Tumor interstitial fluid and postoperative recurrence of tumors: An experimental study for verifying hypothesis of “tumor-phlegm microenvironment”

Objective: To explore a method of extracting tumor interstitial fluid (TIF) which is similar to muddy phlegm in Chinese medicine (CM), interleukin-8 (IL-8) in concentration was taken as the representative of the content of TIF, analyzed in the extracted TIF and the original tumor tissue, and examined to see whether TIF has an interfering effect on tumor recurrence. Methods: Tumor tissue was ground, centrifuged, and filtered for intercellular substances. Tumor-bearing Kunming S180 mice were raised for 21 days and then the tumors were removed to observe the influence of intervention with TIF, normal saline (NS) and a blank control on tumor recurrence. Results: The content of IL-8 in the filtered and unfiltered tumor tissue was not significantly different (P0.05). Postoperative tumor recurrence in TIF intervention group was significantly higher than that in the NS intervention and control groups (60%, 12/20 vs. 20%, 4/20 vs. 15%, 3/20, χ2=11.058, P0.01). Tumor cells grew vigorously and infiltrated to muscular tissue in TIF intervention group. Large numbers of tumor cells were seen necrotic in the NS intervention group, and small numbers of tumor cells were seen necrotic in the blank control group. Conclusions: TIF can be effectively extracted by the means described. It does not contain tumor cells, but its contents such as IL-8 may stimulate tumor cell growth and promote postoperative tumor recurrence, which provided preliminary experimental basis for hypothesis of "tumor-phlegm microenvironment".     

Gastric cancer induced by N—methyl—N‘—nitro—N—nitrosoguanidine in rat with ulcers and expressions of ras and c—erbB2 genes

Objective:To observe the series of pathological changes during the development of gastric adenocarcinoma in ulcerative rats induced by N-methyl-N‘-nitrosoguanidine(MNNG),and the expression profile of related oncogenic protein.Methods:MNNG was administered in rats with ulcers due to acetic acid treatment to induce gastric cancer,and the protein expressions of ras and c-erbB2 genes in the ulcer were examined immunohistochemically along with pathological examination.Results:The incidence of gastric adenocarcinoma in the model group reaches 40%(6/15),while none of the rats developed cancer in the control group with ulcers.Postiive expressions of the proteins of p21 ras and c-erbB2 were observed in the tissues undergoing canceration in the 6 rats of model group,but were not observed in the 5 control rats;p53 protein expression,however,failed to be detected in both groups.Conclusion:A new animal model of gastric cancer has been established in rats with gastric ulcer after MNNG treatment,which may facilitate the pharmacological reserach of gastric cancer.     

Effect of Soothing Gan (Liver) and Invigorating Pi (Spleen) Recipes on TLR4-p38 MAPK Pathway in Kupffer Cells of Non-alcoholic Steatohepatitis Rats

Objective: To investigate the mechanism of inflammatory-mediated toll-like receptor 4(TLR4)-p38 mitogen-activated protein kinase(p38 MAPK) pathway in Kupffer cells(KCs) of non-alcoholic steatohepatitis(NASH) rats and the intervention effect of soothing Gan(Liver) and invigorating Pi(Spleen) recipes on this pathway. Methods: After 1 week of acclimatization, 120 Sprague-Dawley male rats were randomly divided into 8 groups using a random number table(n=15 per group): normal group, model group, low-dose Chaihu Shugan Powder(柴胡疏肝散, CHSG) group(3.2 g/kg), high-dose CHSG group(9.6 g/kg), low-dose Shenling Baizhu Powder(参苓白术散, SLBZ) group(10 g/kg), high-dose SLBZ(30 g/kg) group, and low-and highdose integrated recipe(L-IR, H-IR) groups. All rats in the model and treatment groups were fed with a high-fat diet(HFD). The treatments were administrated by gastrogavage once daily and lasted for 26 weeks. The liver tissues were detected with hematoxylin-eosin(HE) and oil red O staining. Levels of liver lipids, serum lipids and transaminases were measured. KCs were isolated from the livers of rats to evaluate the mRNA expressions of TLR4 and p38 MAPK by real-time fluorescence quantitative polymerase chain reaction, and proteins expressions of TLR4, p-p38 MAPK and p38 MAPK by Western blot. Levels of inflammatory cytokines including tumor necrosis factor α(TNF-α), interleukin(IL)-1 and IL-6 in KCs were measured by enzyme-linked immunosorbent assay. Results: After 26 weeks of HFD feeding, HE and oil red O staining showed that the NASH model rats successfully reproduced typical pathogenesis and histopathological features. Compared with the normal group, the model group exhibited significant increases in body weight, liver weight, liver index, serum levels of total cholesterol(TC), triglyceride(TG), low-density lipoprotein cholesterol, and aspartate aminotransferase as well as TC and TG levels in liver tissues, and significant decrease in serum level of high-density lipoprotein cholesterol(P0.05 or P0.01), while those indices were significantly ameliorated in the H-IR group(P0.05 or P0.01). Higher levels of TNF-α, IL-1 and IL-6 in KCs were observed in the model group compared with the normal group(P0.01). Significant decreases in TNF-α, IL-1 and IL-6 were observed in the H-SLBZ, H-IR and L-IR groups compared with the model group(P0.05 or P0.01). The m RNA expressions of TLR4 and p38 MAPK and protein expressions of TLR4, p38 MAPK and p-p38 MAPK in KCs in the model group were significantly higher than the normal group(P0.01), while those expression levels in the L-IR and H-IR groups were significantly lower than the model group(P0.05 or P0.01). Conclusions: Inflammation in KCs might play an important role in the pathogenesis of NASH in rats. The data demonstrated the importance of TLR4-p38 MAPK signaling pathway in KCs for the anti-inflammatory effect of soothing Gan and invigorating Pi recipes.     

Intervention Effect of Jianxin Decoction（健心汤） on Serum Cytokine Level of Congestive Heart Failure Patients

Objective: To investigate the intervention effect of Jianxin Decoction (健心汤, JXD) on the cytokine level in serum of patients with congestive heart failure (CHF). Methods: Sixty-six patients with CHF were randomly divided into the control group (n=33) and the trial group (n=33). The control group received conventional treatment, and the trial group was treated with conventional therapy plus JXD for 4weeks. Before and after treatment, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and nitrogen monoxide (NO) in serum and cardiac function were determined. Results: After treatment, the levels of TNF-α,IL-6 and NO were significantly lower than those before treatment (P＜0.05, or P＜0. 01) in the two groups,and the lowering degree of the indices in the trial group was more significantly reduced than that in the control group (P＜0. 05). And cardiac functions in both groups were improved significantly (P ＜ 0.05, or P ＜0.01). Conclusion: JXD could prevent and reverse ventricular remodeling so as to ameliorate cardiac function through regulating the levels of cytokines.     

Effects of hydroxy safflower Yellow-A on tumor capillary angiogenesis in transplanted human gastric adenocarcinoma BGC-823 tumors in nude mice

OBJECTIVE:To study the effects of hydroxy safflower yellow A(HSYA) on tumor capillary angiogenesis in transplanted human gastric adenocarcinoma BGC-823 tumors in nude mice.METHODS:BGC-823 cells were injected subcutaneously into the right anterior armpit of nude mice to establish an animal model of transplanted tumors.After 24 h,18 nude mice injected with tumor cells were randomized into model,control,and HSYA 0.028 g/L groups,with six mice in each group.Transplanted tumors were excised on day 20.Tumor inhibition ratios were calculated for the transplanted tumors.Pathological changes and capillary angiogenesis in the tumors were observed by light microscopy.RESULTS:Tumors in the model group grew more quickly than those in the control and HSYA groups,with inhibition ratios of 48% and 30%,respectively.The microvessel count in the HSYA group was lower than in the model group(P<0.01),and microvessel density was also lower in the HSYA group(P<0.05).Pathological changes were more obvious in tumors in the model group compared to the HSYA group.CONCLUSION:HSYA inhibits the growth of transplanted BGC-823 tumors,and its effects on tumor capillary angiogenesis may represent one of the mechanisms responsible for this antineoplastic effect.     

Correlation between elevated FOXP3 expression and increased lymph node metastasis of gastric cancer

Background FOXP3 was thought to express in the T-cell lineage exclusively until recently when FOXP3 was shown to be expressed by cancer cells. It was indicated that FOXP3 may play a wider role in biology by endowing tumor cells with immune suppressive activity. However, researches between FOXP3 and lymph node metastasis of gastric cancer were relatively infrequent, so the present work was aimed to investigate the relationship between FOXP3 expression and lymph node metastasis in human gastric cancer.Methods A total of 122 gastric cancer patients were enrolled in this study, and gastric tumor specimens and lymph nodes were acquired. Thirty patients who had chronic superficial gastritis diagnosed by gastroscopy contemporaneously in the Peking University People＇s Hospital were chosen randomly as the control group. Immunohistochemistry was performed to evaluate FOXP3 expression. A survival analysis on the 122 patients was then performed. Then, NCI-N87cell lines were used to confirm FOXP3 expression in gastric carcinoma cells. Finally, evaluation of FOXP3 expression in gastric tumor and peritumor tissues in 12 patients were conducted using immunohistochemistry and Western blotting. A X2 test or Fisher＇s exact test （bilateral） was conducted to compare the percentage of positive percentage staining between groups. Kaplan-Meier analysis was performed for survival analysis.Results FOXP3 was expressed by gastric cancer cells and peritumor epithelial cells. FOXP3 expression was increased in primary tumors （58.2%） than that in control group （26.7%）. In the lymph-node metastasis group, the incidence of lymph node metastasis which was less than 60% had a significant upregulation of FOXP3 in primary tumors and lymph nodes.However, the frequency of FOXP3 expression had no relationship with survival.Conclusion FOXP3 probably has a relationship with lymph node metastasis of gastric cancer.     

Effects of different therapeutic methods and typical recipes of Chinese medicine on activation of c-Jun N-terminal Kinase in Kupffer cells of rats with fatty liver disease

<正>Objective:To observe the effects of different therapeutic methods and the recipes of Chinese medicine(CM) on the activation of c-Jun N-terminal kinase(JNK) in Kupffer cells of rats with fatty liver disease and to explore the mechanisms of these therapeutic methods.Methods:By using a random number table,98 rats were randomly divided into 7 groups:control group,model group,and 5 treatment groups,including soothing Liver(Gan) recipe group,invigorating Spleen(Pi) recipe group,dispelling dampness recipe group,promoting blood recipe group,and complex recipe group.Rats in the control group were fed with normal food and distilled water by gastric perfusion,while rats in the model group were fed with high-fat food and distilled spirits by gastric perfusion.Rats in the 5 treatment groups were fed with high-fat food and corresponding recipes by gastric perfusion.Twelve weeks later,all rats were sacrificed and liver tissues were stained for pathohistological observation.Kupffer cells were isolated from livers of rats to evaluate JNK and phospho-JNK expressions by Western blotting.Results:The grade of hepatic steatosis was higher in the model group than the control group(P0.05).Compared with the model group,the grade of fatty degeneration in soothing Liver recipe group and invigorating Spleen recipe group were significantly ameliorated(P0.05).Expressions of JNK and phospho-JNK in Kupffer cells were significantly higher in the model group than those in the control group(P0.05,P0.01).Compared with the model group,expressions of JNK in all treatment groups decreased,especially in invigorating Spleen recipe group and promoting blood recipe group(P0.05).Compared with the model group,expressions of phospho-JNK in all treatment groups declined significantly(P0.01),especially in soothing Live recipe group and invigorating Spleen recipe group. Conclusions:The high expressions of JNK and phospho-JNK in Kupffer cells might play an important role in the pathogenesis of fatty liver disease in rats.The recipes of CM,especially invigorating Spleen recipe and soothing Liver recipe,might protect liver against injury by reducing the total JNK protein content and inhibiting the activation of JNK protein in Kupffer cells of fatty liver model rats,which showed beneficial effects on fatty liver disease.     

Clinical values of palliative gastrectomy for late-staged gastric cancer

Objective To investigate the clinical importance of palliative gastrectomy for late-staged gastric cancer. Methods From June 1994 to October 2001,95 patients with late-staged gastric cancer underwent palliative operation. Clinicopathological and prognostic parameters between 64 patients with palliative gastrectomy (PG group) and 31 patients with unresectable operation (UO gruop) was compared retrospectively. Results The age and gender ratioes were not different between the two groups. The incidence of large volume (diameter≥8 cm), serosal invasion (T4) and late TNM stage (Ⅳ stage) were significantly higher in the UO group than that in the PG group. There was no difference in peritoneal disemination, distant lymph node and hepatic metastasis, and tumor location between the two groups. The one- and two-year survival of the patients in the PG group was 48. 1 % and 23.1% ,and significandy better than 13.5% and 0 in the UO group. Conclusion Palliative gastrectomy, compared with unresectable operation, can     

Effects of Ad-p27mt gene transfer on the expression of Bax,Bcl-2, VEGF and MMP-9 in the transplanted liver tumors in nude mice

In this study, the mechanism by which Ad-p27mt inhibits the growth, invasion and metastasis of transplanted liver tumor was studied by examining the effects of Ad-27mt gene transfer on the expression of Bax, Bcl-2, VEGF and MMP-9 in the transplanted liver tumors in nude mice.The model of transplanted hepatic tumor was established in nude mice.The mice were then divided into three groups, which were injected with PBS, Ad-LacZ and Ad-p27mt and the growth of the transplanted liver tumor was observed.The expressions of P27, Bax and Bcl-2 proteins were detected by Western blotting and the expressions of VEGF and MMP-9 were immunohistochemically determined.Our result showed that the tumor size, expressions of Bax, Bcl-2 proteins, VEGF and MMP-9 were all lower than those in PBS and Ad-LacZ groups and the differences were statistically significant (P<0.05).Our study suggested that Ad-p27mt could inhibit the growth, invasion and metastasis of hepatic cancer by lowering the expressions of VEGF and MMP-9.     

Effects of angiotensin-converting enzyme inhibitors and angiotensin II type 1 receptor blockers on lymphangiogenesis of gastric cancer in a nude mouse model

Background Angiotensin-converting enzyme inhibitors （ACEI） and angiotensin II type 1 receptor blockers （ARB） can inhibit tumor growth by inhibition of angiogenesis. This study was designed to study the anticancer effects of ACEI and ARB on tumor growth and lymphangiogenesis in an implanted gastric cancer mouse model. Methods A model of gastric cancer was established by subcutaneously inoculating human gastric cancer cell line SGC-7901 into 60 nude mice. One week later, all mice were randomly divided into 5 groups. A control group received physiologic saline once daily for 21 days. Mice in the 4 treatment groups received one of the following agents by gavage once daily for 21 days： perindopril, 2 mg/kg; captopril, 5 mg/kg; Iosartan, 50 mg/kg; or valsartan, 40 mg/kg. Twenty-one days after treatment, all the mice were sacrificed and the tumors were removed. Tumor sections were processed, and immunohistochemical methods were used to observe the expressions of vascular endothelial growth factor C （VEGF-C）, matrix metalloproteinase 7 （MMP-7）, and lymphatic microvessel density （LMVD）.
Results Tumor volume was significantly inhibited in all ACEI and ARB groups, compared with the control group （all P 〈0,01）. LMVD in the ACEI and ARB groups was also significantly lower than that of the control group （all P 〈0.01）. In the ACEI groups, the expressions of VEGF-C and MMP-7 were both significantly decreased, compared with the control group （all P 〈0.05）. In the ARB groups, expression of VEGF-C was significantly decreased compared with the control group （all P 〈0.05）. However, no significant difference was found in the expression of MMP-7 between ARB groups and the control group.
Conclusion In a mouse model, ACEI and ARB might inhibit gastric cancer tumor growth by suppressing lymphangiogenesis.     

EXPRESSION OF MASPIN AND KAI1 AND THEIR CLINICOPATHOLOGICAL SIGNIFICANCE IN CARCINOGENESIS AND PROGRESSION OF GASTRIC CANCER

Objective To investigate the roles of maspin and kail expression in tumorigenesis and progression of gastric cancer.Methods Maspin and kail expressions were detected in normal gastric mucosa (n = 182), gastric dysplasia (n = 69), and gastric cancer (n = 113) by immunohisto-chemistry. Their expressions were compared with clinicopathological parameters of tumors. Relationship between maspin and kai1 expression was also concerned in gastric cancer.Results The positive rates ofmaspin expression were 79.8% (145/182), 75.4% (52/69), and 50.4% (57/113) in normal gastric mucosa, gastric dysplasia, and gastric cancer, while those of kail expression were 81.9% (149/182), 65.2% (49/69),and 58.4% (66/113) in corresponding tissues respectively. Gastric cancer less frequently expressed maspin than the normal gastric mucosa and gastric dysplasia (P ＜ 0.05), while dysplasia and cancer showed less frequent expression of kail than normal mucosa (P ＜ 0.05). Maspin expression showed negative association with invasive depth, metastasis, Lauren's and histological classifications (P ＜ 0.05), but not with tumor size, Borrmann's classification, growth pattern or TNM staging (P ＞ 0.05). Kail expression was negatively correlated with invasive depth, metastasis, growth pattern, Lauren's and histological classifications (P ＜ 0.05), but not with tumor size, Borrmann's classification or TNM staging (P ＞ 0.05). Maspin and kail were collaboratively expressed in gastric cancer (P ＜ 0.05).Conclusions Down-regulated expressions of maspin and kail play an important role in gastric carcinogenesis. Abnormal expression of maspin and kail might have inhibitory effects on invasion and metastasis of gastric cancer and act as an effective and objective marker to indicate the pathobiological behaviors of gastric cancer.     

Effect of p27 Gene Combined with Pientzehuang (片仔癀) on Tumor Growth in Osteosarcoma-Bearing Nude Mice

Objective:To observe the effect of p27 gene recombinant adenovirus combined with Chinese medicine Pientzehuang(片仔癀) on the growth of xenografted human osteosarcoma in nude mice.Methods:Tissue transplantation was used to construct the orthotopic model of human osteosarcoma Saos-2 cell in nude mice.Thirty tumor-bearing nude mice were randomly divided into 5 groups with 6 mice in each group:blank control group(model of osteosarcoma),empty vector group(recombinant adeno-associated virus-multiple cloning site),Pientzehuang group,p27 gene group and combined treatment group(p27 gene combined with Pientzehuang).The effect of combined treatment on human osteosarcoma was analyzed through the tumor formation,tumor volume and inhibition rate of tumor growth.The expression of p27 was measured by immunohistochemical staining and Western blot.Results:The orthotopic model of osteosarcoma in nude mice was successfully constructed.The general appearance of tumor-bearing nude mice in Pientzehuang and p27 gene groups was markedly improved compared with the blank control group;and in the combined treatment group it was significantly improved compared with the Pientzehuang and p27 gene groups.The tumor growth in the Pientzehuang and p27 gene groups was significantly inhibited compared with the blank control group(P0.05);while in the combined treatment group it was markedly inhibited compared with the Pientzehuang and p27 gene groups(P0.05).The rates of tumor growth inhibition were 34.1%,56.5%and 63.8%in the Pientzehuang,p27 gene and combined treatment groups,respectively.Meanwhile,the protein expression of p27 gene in the p27 gene group was significantly increased compared with the blank control group(P0.05);and it was significantly increased in the combined treatment group compared with the p27 gene and Pientzehuang groups(P0.05).Conclusion:p27 gene introduced by adenovirus combined with Pientzehuang can inhibit the growth of human osteosarcoma cell Saos-2 in nude mice.     

Effects of Jinmaitong Capsule (筋脉通胶囊) on Ciliary Neurotrophic Factor in Sciatic Nerves of Diabetes Mellitus Rats

Objective:To study the effects of the Chinese medicine Jinmaitong Capsule(筋脉通胶囊,JMT) on the pathomorphology of sciatic nerves,ciliary neurotrophic factor(CNTF),and the mRNA expressions of CNTF in rats with streptozotocin-induced diabetes mellitus(STZ-DM).Methods:The animal model was established by one time intraperitoneal injection of streptozotocin.The rats were simply divided by random into 5 groups including model group,low-dose JMT group(JL),medium-dose JMT group(JM),high-dose JMT group(JH) and neurotropin group.For each of the above 5 groups,a group of 10 normal Wistar rats matched in body weight,age and gender were set as normal group.Intragastric administrations were started after the animal model established.The JL group were administered with five times the JMT dose recommended for a human adult;the JM group were administered with ten times the JMT dose recommended for a human adult;the JH group were administered with twenty times the JMT dose recommended for a human adult.The neurotropin group was administered with ten times the neurotropin dose recommended for a human adult.All rats were given intragastric administration for 16 weeks and then killed.In the 4th,8th,12th,16th week,body weight and blood glucose level were detected before and after the intervention.The morphologic changes of the sciatic nerves were observed by optical microscope and transmission electron microscope.The CNTFmRNA expressions were detected by real-time fluorescent quantitative polymerase chain protein,and the CNTF protein expressions were detected by immunohistochemical method.Results:The blood glucose levels of the STZ-DM rats were much higher than normal group(P<0.01),and there was no apparent difference between any treatment groups and the model group(P>0.05).Before and after the intervention in the 4th, 8th,12th,16th week,there were no significant differences in the body weight among all the groups(P>0.05). The sciatic nerves of STZ-DM rats might have pathomorphological changes in axons,myelin sheaths,and interstitium.The levels of CNTF and CNTF-mRNA expressions in the STZ-DM rats were both significantly decreased(P<0.01).The sciatic nerves of STZ-DM rats might have pathomorphological changes in axons, myelin sheaths,and interstitium.Conclusion:JMT could improve the pathomorphology of sciatic nerves by increasing CNTF’s and CNTF-mRNA expressions in sciatic nerve tissues,and promote the repair and regeneration of damaged nerve fibers.     

Antioxidant Mechanism of Xiaojin Pill(小金丸) for Treatment of Peyronie's Disease in Rats Based on Matrix Metalloproteinases

Objective: To evaluate the effects of Xiaojin Pill(小金丸) in the treatment of Peyronie's disease(PD) in a rat model. Methods: Twenty-four male Sprague-Dawley rats were randomly divided into four groups with 6 in each: sham operation, PD model, vehicle control and Xiaojin Pill groups. The rats in the sham operation group received penile tunica albsginea(TA) injection with 50 μL vehicle, while the rats in the other 3 groups received 50 μL penile TA injection of 50 μg transforming growth factor(TGF)-β1. Forty-two days after the injection, rats in the vehicle control and Xiaojin Pill groups received 0.5 mL water and Xiaojin Pill solution(107 mg/kg of body weight), respectively by gavage for 28 days, while those in the sham operation and PD model groups did not receive any intervention. After intervention, the expressions of matrix metalloproteinase 2/9(MMP2/9), nitric oxidesynthase(NOS), superoxide dismutase(SOD) and malondialdehyde(MDA) were measured. Results: Rats in the PD model and vehicle control groups presented obvious fibrosis in corpus cavernosum(CC) and demonstrated a significantly increased expressions of MMP2 and MMP9 in the CC compared with the sham operation group(all P0.01). In contrast, the expressions of MMP2 and MMP9 in the Xiaojin Pill group were significantly down-regulated(both P0.01). In addition, the levels of NOS and MDA in CC were significantly increased while the activity of SOD was decreased in the PD model and vehicle control groups compared with the sham operation group(al P0.01). After Xiaojin Pil treatment, the levels of MDA, NOS and SOD appeared to be corrected(al P0.01). Conclusions: Xiaojin Pill could reduce fibrosis in the CC by decreasing the expressions of MMPs, NOS and MDA, and by increasing the activity of SOD. Therefore, Xiaojin Pill might be a therapeutic option for PD.     

Effects of Propofol on The mRNA Expression of Toll-like Receptor 4 in BV-2 Cells During Mimic Ischemia-Reperfusion Injury In Vitro

This study investigated the effects of propofol on the mRNA expression of Toll-like receptor-4 (TLR4) in BV-2 cells during mimic ischemia-reperfusion (I/R) injury in vitro. BV-2 cells, a mouse microglia line, were cultured and divided into 4 groups at random: control group (group C), ischemia/reperfusion group (group I/R), low-dose propofol (25 μmol/L) intervention group (group PF25) and high-dose propofol (100 μmol/L) intervention group (group PF100). The mRNA expression of TLR4 and NF-κB was measured by means of RT-PCR. TNF-α levels in the supernatants of BV-2 cells were detected by ELISA. The results showed that the mRNA expression of TLR4 and NF-κB was significantly higher in groups I/R, PF25 and PF100 than in group C (P<0.01). And the TNF-α level in the supernatants was elevated in groups I/R, PF25 and PF100 as compared with that in group C (P<0.01). After pre-treatment with propofol, the mRNA expressions of TLR4 and NF-κB and the TNF-α level were significantly decreased in groups PF25 and PF100 in comparison to those in group I/R (P<0.01). And the decrease in those indicators was more significant in group PF100 than in group PF25 (P<0.01). It was concluded that propofol exerted brain-protecting effects during I/R injury by suppressing the mRNA expressions of TLR4 and NF-κB and deceasing the TNF-α level.     

Modulation of expression of P16 and Her2 in rat breast tissues of mammary hyperplasia model by external use of Rupifang Extract

OBJECTIVE:To observe the effect of Rupifang Extract in external use on expression of proto-oncogenes her2 and tumor suppression genes p16 in rat breast tissues of mammary hyperplasia model.To explore the mechanisms of Rupifang Extract in external use for preventing and treating mammary hyperplasia.METHODS:Thirty virginal female Wistar rats were randomized into 5 groups,6 in each,A:blank control group;B:model group;C:the low dose group of Rupifang;D:the middle dose group of Rupifang;and E:The high dose group of Rupifang.The mammary hyperplasia rat models were produced by injecting estradiol benzoate and progesterone and irritating by tail nipping.Drug intervention was also launched during the model formation.After 30 days,the expression of her2 and p16 in breast tissues of rats in each group were detected by the SP immunohistochemical method.RESULTS:Compared with Blank control group,the expression of her2 in breast tissues in Model group was higher,and the expression of p16 was lower(P<0.05 or P<0.01).After intervention with Rupifang Extract,compared with Model group,the expression of her2 in breast tissues in Rupifang groups was lower,and the expression of p16 higher(P<0.05 or P<0.01).CONCLUSION:The mechanisms of Rupifang Extract in external application for preventing and treating mammary hyperplasia may be reducing the expression of proto-oncogenes her2 and increasing the expression of tumor suppression genes p16.     

Telomerase activity analysis of esophageal carcinoma using microdissection-TR AP assay

Objectives To investigate telomerase activity in esophageal squamous cell carcinoma (SCC) and its preneoplasia lesions, and to study the relationships between telomerase activity and cancer differentiation, cancer invasiveness, and lymphatic metastasis.Methods Telomerase activity in esophageal SCC tissues, adjacent dysplasia tissues and normal epithelia from the surgical edge were assessed by microdissection-TRAP elomeric repeat amplification protocol）-silver staining assay.Results Telomerase activity was detected in 37 (82.2%) of 45 esophageal tumors, 23 (79.3%) of 29 dysplasias, and 2 (5%) of 40 normal epithelia.There was a significant difference in activity between dysplasia and normal epithelium, as well as between tumor and normal epithelium.Twenty-six (92.9%) of 28 tumors with lymphatic metastasis had detectable telomerase activity compared to 11 (64.7%) of 17 non-lymphatic metastasis tumors.These relationships were statistically significant (P&lt;0.05), but the one between telomerase activity and tumor grade was not.Conclusion Telomerase activity was high both in esophageal SCC and their preneoplasia lesions.The telomerase activity in SCC tissue was related to lymphatic metastasis, but not to cancer differentiation.     

Preliminary research on the pathological role of cathepsin-B in subcutaneous heteroplastic pancreatic carcinoma in nude mice

Background Cathespin-B （cath-B） is an important proteolytic enzyme involved in the disease course of invasion in many types of cancer. Cath-B expression in subcutaneous heteroplastic pancreatic carcinoma in nude mice has not been studied. We investigated the role of cath-B in a model of heteroplastic pancreatic carcinoma in BALB/c nude mice. Methods Thirty-two six-week-old female BALB/c nude mice were equally divided into four groups. PANC-1 cells were inoculated subcutaneously in the left axillary region. Besides volume, weight of subcutaneous tumor, and change in body weight, cath-B expression in each group was measured by immunohistochemical staining, PCR and Western blotting. Its relationship to microvessel density （MVD）, CD44v6, and placenta growth factor （PLGF） was also examined. CA-074Me, a specific inhibitor of cath-B, was injected intraperitoneally （i.p.） at different stages of tumor growth in group B and C. Gemcitabine （GEM）, was also injected （i.p.） in group D to compare anti-tumor efficacy with CA-074Me. Results Expression of cath-B at different levels was related to tumor growth, MVD, and PLGF expression. In group A （control group）, cath-B expression was enhanced more than that seen in other groups. CA-074Me clearly inhibited cath-B expression and tumor growth in group B. There was no difference between group C and D with respect to anti-tumor effect. Conclusions Cath-B correlates with the growth and angiogenesis of tumors, but not with the adhesion induced by CD44v6. CA-074Me clearly inhibited cath-B expression and demonstrated an anti-neoplastic and anti-angiogenesis effect.