Isotype distribution of anti-Ro/SS-A and anti-La/SS-B antibodies in plasma and saliva of patients with Sjögren's syndrome

In this study an ELISA assay was used to quantify the levels of antibodies against the recombinant Ro 52 kD, Ro 60 kD, and La 48 kD proteins in plasma and saliva of 17 Sj?gren's syndrome patients. The levels of total IgG, IgA, and IgM were also quantified. About one third of the patients had salivary enrichment of IgA and IgM against the Ro 52 kD, Ro 60 kD, and La 48 kD antigens and IgG against La 48 kD, while no enrichment of IgG against Ro 52 kD and Ro 60 kD was found. Most correlations between plasma and saliva levels of antigen specific antibodies were highly significant, as were most correlations between focus score and levels of antigen specific antibodies in plasma and saliva. In total, the results support the hypothesis that autoantibodies are produced in the salivary glands. The strong correlation between focus score and plasma and saliva levels of autoantibodies, indicates that the local autoantibody production is a consequence of local inflammation.     

Molecular study and genotype/phenotype correlation of β Thalassemia in Malaysia

Introduction: To study the ß‐gene mutations spectrum, the genotype/phenotype correlation, the modulatory effect of co‐inherited factors such as α‐gene mutations and of Xmn1 polymorphism in a large cohort of Malaysian patients. Methods: A total of 264 cases clinically diagnosed as Thalassemia major (TM) (111), Thalassemia intermedia (21), HbE‐β Thalassemia (131), and 1 HbE homozygous were studied. The detection of α and ß gene mutations and characterization of Xmn1 polymorphism were performed by multiplex PCR, amplification refractory mutation system (ARMS), DNA sequencing, and restriction fragment length polymorphism (RFLP)‐PCR. Results: A total of 19 ß Thalassemia mutations were characterized. CD26 and CD41/42 were the most common found in the Malay and Chinese population, respectively. The sensitivity of the clinical diagnosis for β TM, thalassemia intermedia, and HbE/β thalassemia was 94.0%, 15.2%, and 89.2%, respectively. Patients with Xmn1 heterozygosity [+/?] required less frequent transfusion compared with those without the polymorphism. Co‐inheritance of α‐thalassemia alleviates the severity of HbE‐β thalassemia in our cohort. Conclusion: Molecular analysis should be used for a better diagnosis and management of β thalassemia.     

C/EBPβ and CHOP participate in Tanshinone IIA-induced differentiation and apoptosis of acute promyelocytic leukemia cells in vitro

Our studies indicated that Tanshinone IIA (TanIIA), which is widely applied in the treatment of cardiovascular diseases with a rare occurrence of side effects, could promote APL cell differentiation and apoptosis. We found TanIIA induced the differentiation of NB4 and MR2 cells with elevated C/EBPβ and CHOP. When C/EBPβ was overexpressed in NB4 cells, the level of CD11b in the transfected cells was significantly elevated. When we used CHOP siRNA to suppress CHOP expression in NB4 cells and then treated these cells with a high concentration of TanIIA, the differentiation and apoptosis of these cells were both significantly increased. These data demonstrate that C/EBPβ is critical for APL cell differentiation and apoptosis induced by TanIIA, and that CHOP acts as a negative regulator of C/EBPβ activity. Our study suggested that TanIIA is a promising drug for treating newly diagnosed and ATRA-resistant APL, and a high concentration of TanIIA associated with inhibition of CHOP, maybe a potentially promising therapy strategy.     

Prognostic and predictive significance of MSI in stages Ⅱ/Ⅲ colon cancer

In colon cancer,classic disease staging remains the key prognosis and treatment determinant.Although adjuvant chemotherapy has an established role in stageⅢcolon cancer patients,in stageⅡit is still a subject of controversy due to its restriction to a small subgroup of patients with high-risk histopathologic features.Patients with stageⅡtumors form a highly heterogeneous group,with five-year relative overall survival rates ranging from 87.5%(ⅡA)to 58.4%(ⅡC).Identifying those for whom adjuvant chemotherapy would be appropriate and necessary has been challenging,and prognostic markers which could serve in the selection of patients more likely to recur or benefit from adjuvant chemotherapy are eagerly needed.The stronger candidate in this category seems to be microsatellite instability(MSI).The recently reported European Society for Medical Oncology guidelines suggest that MSI should be evaluated in stageⅡcolorectal cancer patients in order to contribute in treatment decisionmaking regarding chemotherapy administration.Thehypothetical predictive role of MSI regarding its response to 5-fluorouracil-based adjuvant chemotherapy has proven a much more difficult issue to address.Almost every possible relation between MSI and chemotherapy outcome has been described in the adjuvant colon cancer setting in the international literature,and the matter is far from being settled.In this current report we critically evaluate the prognostic and predictive impact of MSI status in patients with stageⅡand stageⅢcolon cancer patients.     

Past and present of parasitology in China

Comparison was made of the parasitological institutions, personnel and work betweenthe periods before and after the founding of the People's Republic. There were 3 nationalinstitutions concerned with parasitology before the 50s, i.e. the Institute of TropicalMedicine of Zhejiang Province, the Fan Memorial Institute of Biology and the CentralHealth Station which was the precursor of the National Institute of Health. In contrast,     

Assessment of resin perfusion in hepatic failure in vitro and in vivo

AIM:To observe the adsorbent effect of resin on endotoxin,cytokine,bilirubin in plasma of patients with hepatic failureand to determine the resin perfusion as an artificial liversupport system in the treatment of hepatic failure.METHODS:One thousand milliliters of discarded plasmawas collected from each of 6 severe hepatitis patients treatedwith plasma exchange.The plasma was passed through aresin perfusion equipment for 1-2 h via extracorporealcirculation,and then absorbent indicators of transaminase,bilirubin,blood ammonia,endotoxin and cytokines wereexamined.In the meantime,study of in vivo resin plasmaperfusion was performed on 7 severe hepatitis patients tocompare the changes of endotoxin and cytokines in bloodbefore and after perfusion.RESULTS:The levels of total bilirubin,endotoxin,interleukin1β and TNF-α in plasma were significantly decreased afterin vitro resin plasma perfusion.The levels of interleukin 1β,TNF-α and endotoxin in blood were also evidently declinedafter in vivo resin plasma perfusion.Nevertheless,no obviouschanges in IL-6,creatinine (Cr) and urea nitrogen (UN),bloodammonia and electrolytes were found both in vitroand in vivo.CONCLUSION:Bilirubin,endotoxin and cytokines in plasmaof patients with hepatic failure can be effectively adsorbedby resin in vitro.Most cytokines and endotoxin in plasma canalso be effectively removed by resin in vivo.It demonstratesthat resin perfusion may have good treatment efficacy onhepatic failure and can be expected to slow down theprogression of hepatic failure.     

Alcohol metabolites and lipopolysaccharide： Roles in the development and/or progression of alcoholic liver disease

The onset of alcoholic liver disease (ALD) is initiated by different cell types in the liver and a number of different factors including: products derived from ethanol- induced inflammation, ethanol metabolites, and the indirect reactions from those metabolites. Ethanol oxidation results in the production of metabolites that have been shown to bind and form protein adducts,and to increase inflammatory, fibrotic and cirrhotic responses. Lipopolysaccharide (LPS) has many deleterious effects and plays a significant role in a number of disease processes by increasing inflammatory cytokine release. In ALD, LPS is thought to be derived from a breakdown in the intestinal wall enabling LPS from resident gut bacterial cell walls to leak into the blood stream. The ability of adducts and LPS to independently stimulate the various cells of the liver provides for a two-hit mechanism by which various biological responses are induced and result in liver injury. Therefore,the purpose of this article is to evaluate the effects of a two-hit combination of ethanol metabolites and LPS on the cells of the liver to increase inflammation inflammation and fibrosis, and play a role in the development and/or progression of ALD.     

Detection of bcl-2 and bax expression and bcl-2/JH fusion gene in intrahepatic cholangiocarcinoma

AIM:To investigate the relationship between bcl-2 gene and its related protein bax and intrahepatic cholangiocellular carcinoma(CCC). METHODS:Semi-nested in situ PCR(SNISPCR)and imm- unohistochemistry were performed to detect bcl-2/JH fusion gene and bcl-2,bax protein expression in 29 cases of CCC. RESULTS:No bcl-2/JH fusion gene was found in all cases of CCC,72.4% of 29 cases expressed bcl-2 protein.Bcl-2 protein expression was related to histopathological grades (P<0.05).There was no corresponding relationship between bcl-2/JH fusion gene formation and bcl-2 protein expression in CCC(P<0.05).Bax was expressed in 10.3% of 29 cases. The ratio of bcl-2 to bax in normal liver tissues(3.5 to 1)was different from that in tumor tissues(7.0 to 1). CONCLUSION:It is suggested that bcl-2/JH fusion gene formation is not a frequent event and may not play an important role in the pathogenesis of CCC.However,aberrant ratio of bcl-2 to bax protein expression may be involved in the course of tumorigenesis of CCC.Abnormal bcl-2 protein expression may not be solely resulted from bcl-2/JH fusion gene.     

Efficacy and Safety of Alirocumab in Patients with Heterozygous Familial Hypercholesterolemia and LDL-C of 160 mg/dl or Higher

Purpose

Even with statins and other lipid-lowering therapy (LLT), many patients with heterozygous familial hypercholesterolemia (heFH) continue to have elevated low-density lipoprotein cholesterol (LDL-C) levels. ODYSSEY HIGH FH (NCT01617655) assessed the efficacy and safety of alirocumab, a proprotein convertase subtilisin/kexin type 9 monoclonal antibody, versus placebo in patients with heFH and LDL-C ≥ 160 mg/dl despite maximally tolerated statin ± other LLT.

Methods

Patients were randomized to subcutaneous alirocumab 150 mg or placebo every 2 weeks (Q2W) for 78 weeks. The primary endpoint was percent change in LDL-C from baseline to week 24.

Results

Mean baseline LDL-C levels were 196.3 mg/dl in the alirocumab (n = 71) and 201.0 mg/dl in the placebo groups (n = 35). Significant mean (standard error [SE]) reductions in LDL-C from baseline to week 24 were observed with alirocumab (?45.7 [3.5] %) versus placebo (?6.6 [4.9] %), a difference of ?39.1 (6.0) % (P < 0.0001). Absolute mean (SE) LDL-C levels were reduced from baseline by 90.8 (6.7) mg/dl with alirocumab at week 24, with reductions maintained to week 78. Treatment-emergent adverse events were generally comparable between groups. Injection-site reactions were more frequent in the alirocumab group (8.3 %) versus placebo (5.7 %); most were mild in severity and did not result in study medication discontinuation.

Conclusions

In patients with heFH and very high LDL-C baseline levels despite maximally tolerated statin ± other LLT, alirocumab 150 mg Q2W demonstrated significant reductions in LDL-C levels with 41 % of patients achieving predefined LDL-C goals. Alirocumab was generally well tolerated.

     

Overexpression of β3/γ2 chains of laminin-5 and MMP7 in biliary cancer

AIM： To clarify the clinicopathological significance of laminin-5 y2 （LNγ2） and 133 （LNI33） chains and MMP7 expression in biliary tract cancer. METHODS： We analyzed the association between immunohistochemically detected LNγ2, LNβ3, and MMP7 expression in biliary tract cancer and clinicopathological characteristics. Activity of MMP7 was analyzed by casein zymography. An in vitro invasion assay after treatment with MMP7-specific siRNA was performed. RESULTS： LNγ2 expression was predominantly observed in carcinoma cells at the invasive front. LNγ2 expression was seen in 57% of patients with biliary tract cancer, and was associated with depth of invasion, histologic type, and advanced stage. The expression pattern of LNβ3 was classified Into two types： invasive front dominant type （38%） and diffuse type （28%）.The invasive front dominant type was associated with histologic type and advanced stage. MMP7 positivity was correlated with LNγ2 or LNβ3 expression but not with clinicopathological characteristics. Active MMP7 detected by casein zymography was correlated with depth of invasion and advanced stage. Downregulation of MMP7 expression by siRNA resulted in a significant decrease in biliary tract cancer cell invasion in vitro.CONCLUSION： Our results suggest that LNγ2 and LNβ3, in conjunction with MMP7, play a key role in the progression of biliary tract cancer.     

Efficacy and Safety of Ombitasvir/Paritaprevir/Ritonavir and Dasabuvir ± Ribavirin for HCV in Brazilian Adults with Advanced Fibrosis

Introduction and aim. Approximately 650,000 people in Brazil have chronic hepatitis C virus (HCV) infection. We evaluated the safety and efficacy of ombitasvir (OBV)/paritaprevir (PTV)/ritonavir (r) plus dasabuvir (DSV) with/without ribavirin (RBV) in an open-label multicenter phase 3b trial in treatment-naive or interferon (IFN) treatment-experienced Brazilian patients with advanced hepatic fibrosis (METAVIR F3/4) and HCV genotype (GT) 1 infection.Material and methods. All patients received coformulated OBV/ PTV/r daily + DSV twice daily (3-DAA). GTIa-infected patients received 3-DAA plus RBV for 12 weeks, except for prior pegIFN/ RBV nonresponders with cirrhosis who were treated for 24 weeks. GTIb-infected patients received 3-DAA alone (F3) or in combination with RBV (F4) for 12 weeks. The primary endpoint was sustained virologic response (HCV RNA < 15 IU/mL) at post-treatment Week 12 (SVR₁₂).Results. The study enrolled 222 patients, 214 achieved an SVR₁₂ (96.4%; 95% CI, 93.1-98.2%), one GT1a-infected patient experienced virologic breakthrough, six (5 GT1a) relapsed, and one was lost to follow-up. SVR₁₂ was achieved in 111/ 112 (99.1%) GT1b-infected patients, including 42/43 (97.7%) noncirrhotic, and 69/69 (100%) cirrhotic patients; and in 103/110 (93.6%) GT1a-infected patients, including 44/46 (95.7%) noncirrhotic and 59/64 (92.2%) cirrhotic patients. Overall there was a low rate of serious adverse events (n = 6, 2.7%). One patient experienced a treatment-related serious adverse event and one patient discontinued treatment because of an adverse event.Discussion. The results confirm that the 3-DAA regimen with/without RBV is well tolerated and had a favorable safety profile and is efficacious in GT1-infected patients with advanced fibrosis (METAVIR F3/4).     

Mutational Analysis of WTX Gene in Wnt/ β-Catenin Pathway in Gastric, Colorectal, and Hepatocellular Carcinomas

A recent study of Wilms’ tumors discovered a new X chromosome gene, Wilms’ tumor gene on the X chromosome (WTX), which was found to harbor small deletions and point mutations. WTX protein negatively regulates Wnt/ β-catenin signaling, and is considered a tumor-suppressor gene. One of the questions about the WTX gene is whether the genetic alterations of the WTX gene are specific to only Wilms’ tumors. To see whether somatic point mutations of WTX occur in other malignancies, we analyzed the WTX gene for the detection of mutations in 141 cancer tissues by a single-strand conformation polymorphism assay. The cancer tissues consisted of 47 gastric adenocarcinomas, 47 colorectal adenocarcinomas, and 47 hepatocellular carcinomas. Overall, we detected one WTX mutation in the colorectal carcinomas (1/47; 2.1%), but there was no WTX mutation in other cancers analyzed. The detected mutation was a missense mutation (c. 1117G > A (p.Ala373Thr)). Although the WTX mutation is common in Wilms’ tumors, our data indicate that it is rare in colorectal, gastric, and hepatocellular carcinomas. The data also suggest that deregulation of Wnt/ β-catenin signaling by WTX gene mutation may be a rare event in the pathogenesis of colorectal, gastric, and hepatocellular carcinomas.     

Inhibitory effect of dimeric β peptide on the recurrence and metastasis of hepatocellular carcinoma in vitro and in mice

AIM：To block the adhesion of tumor cells to the extracellular matrix, and prevent tumor metastasis and recurrence, the dimer of the β peptide （DLYYLMDLSYSMKG- GDLYYLMDLSYSMK, β2） was designed and synthesized and its anti-adhesion and anti-invasion effects on hepa- tocellular carcinoma cells were assessed. Additionally, its influence on the metastasis and recurrence of mouse hepatocellular carcinoma was measured.
METHODS：The anti-adhesion effect of β2 on the highly metastatic hepatocellular carcinoma cell line HCCLM6 cells and fibronectin （FN） was assayed by the MTT as- say. The inhibition of invasion of HCCLM6 cells by β2 was observed using a Transwell （modified Boyden chamber） and matrigel. Using the hepatocellular carcinoma metas- tasis model and LCI-D20 nude mice, the influence of β2 on the metastasis and recurrence of hepatocellular carci- noma after early resection was investigated.
RESULTS：HCCLM6 cells co-incubated with 100 mmol/L, 50 mmol/L, 20 mmol/L or 10 mmol/L β2 for 3 h showed an obvious decrease in adhesion to FN. The adhesion inhibition ratios were 11.8%, 21.7%, 29.6% and 48.7%, respectively. Additionally, HCCLM6 cells cultured with 100 mmol/L β2 had a dramatic decrease in cell invasion. β2 was also observed to inhibit the incisal edge recur- rence and the distant metastasis of nude mice hepato- cellular carcinoma after early resection （P 〈 0.05）.
CONCLUSION：The β2 peptide can specifically block the adhesion and invasion of HCCLM6 cells, and can inhibit HCC recurrence and metastasis of LCI-D20 model pos-thepatectomy in vivo. Thus, β2 should be further studied as a new anti-tumor drug.     

The Re-emergence and Control of Plague in Yunnan

云南鼠疫流行历史悠久，１９５６年流行被控制后，经调查证实存在家、野两型鼠疫自然疫源地。１９８２年起家鼠鼠疫开始复燃，到１９９６年底，全省已先后从３３个县市检出鼠疫菌，８个县查出鼠疫血凝抗体（ＩＨＡ），９个县市发现鼠疫放免（ＲＩＰ）阳性。其中１９个县市发生人间腺鼠疫病人２２６例，死亡２例。经努力防治，避免了鼠疫的大流行及肺鼠疫的发生，但鼠疫仍以年均增加３个县的速度继续复燃。先后从４种鼠、６种蚤、１种蜱及患者检出鼠疫菌。经调查研究鼠疫的主要宿主是黄胸鼠（Ｒａｔｕｓｆｌａｖｉｐｅｃｔｕｓ），主要传播媒介是印鼠客蚤（Ｘｅｎｏｐｓｙｌａｃｈｅｏｐｉｓ）。鼠疫菌不发酵甘油、发酵麦芽糖，提取出８种分子量质粒，同时还发现有规范化质粒单个缺失菌株。质粒图谱类型和生化特性与分离宿主、媒介和年代无关而有明显的地理分布特征。鼠疫病人中腹股沟淋巴腺肿占６０６２％，３０岁以下年龄组占６２８３％，农民占９２４８％，汉族占７０３５％。传统使用抗菌素及中草药治疗病人有效，但药敏试验以羧苄青霉素、复方新诺明最敏感，链霉素排到第１１位。采取建立鼠疫监测网、早期发现疫情、灭鼠、灭蚤和疫区处理等综合防治措施。目前缺乏有效的免疫菌苗，经济的灭     

Role of TGF-β1/Smads pathway in carotid artery remodeling in renovascular hypertensive rats and prevention by Enalapril and Amlodipine

Objective To investigate the role of transforming growth factor-β1 (TGF-β1), Smad2/3 and Smad7 expressions in carotid artery remodeling in renovascular hypertensive rats, and also the therapeutic effect of Enalapril and Amlodipine. Methods The renovascular hypertensive rat (RHR) models with “two-kidney and one-clip” were established, including model group (n = 6), sham-operated group (n = 6), Enalapril group (10 mg/kg per day, n = 6), Amlodipine group (5 mg/kg per day, n = 6) and combination group (Amlodipine 2.5 mg/kg per day + Enalapril 5mg/kg per day, n = 6). The medication were continuous administrated for six weeks. Carotid artery morphological and structural changes in the media were observed by HE staining, Masson staining and immuno histochemical staining. Media thickness (MT), MT and lumen diameter ratio (MT/LD), and the expression levels of media α-smooth muscle actin (α-actin), proliferating cell nuclear antigen (PCNA), TGF-β1, phosphorylated Smad2/3 (p-Smad2/3) and Smad7 in carotid arteries were measured. Results The media of carotid arteries in RHR model group was significantly thickened, the volume of smooth muscle cell was increased, and the array was in disorder; MT, MT/LD, the proliferation index of smooth muscle cell and collagen fiber area percentage of carotid arteries in the model group were significantly higher than those in the sham-operated group (P < 0.01). Compared to sham-operated group, the model group had significantly higher expressions of TGF-β1 and p-Smad2/3 (P < 0.05) and lower Smad7 expression. Both Enalapril and Amlodipine improved smooth muscle hypertrophy and collagen deposition, reduced RHR carotid MT, MT/LD, proliferation index of smooth muscle cell, collagen fiber area percentage and the expressions of TGF-β1 and p-Smad2/3 (P < 0.05), increased Smad7 expression (P < 0.05). Moreover, the combination treatment of Enalapril and Amlodipine had significantly better effects than single Amlodipine group (P < 0.05), but not single Enalapril group. Conclusions TGF-β1/Smads pathway may participate in the mechanism of carotid artery remodeling in RHR; the role of Amlodipine and Enalapril in inversing carotid artery remodeling may be related to the change of TGF-β1/Smads pathway, the combination treatment of Amlodipine and Enalapril had better effects than single administration of Amlodipine.     

Combined effects of Cantide and chemotherapeutic drugs on inhibition of tumor cells＇ growth in vitro and in vivo

AIM: To investigate the combination effect of hTERT antisense oligonucleotide "Cantide" and three chemotherapeutic drugs (cisplatin, 5-fluorouracil (5-FU) and adriamycin (ADM)) on inhibiting the proliferation of HepG2, BGC and A549 cell lines in vitro, and to investigate the efficacy of Cantide used in combination with cisplatin (DDP) in vivo. METHODS: Cantide was transfected into these tumor cells by Lipofectin, and cell growth activity was calculated by microcytotoxicity assay. In vivo study, cells of HepG2 were implanted in Balb/c nude mice for 4 d. Then Cantide, DDP and Cantide+DDP were given intra peritonea Ily for 24 d respectively. The body weights of the tumor-bearing animals and their tumor mass were measured later to assess the effect of combination therapy in the nude mice. To evaluate the interaction of Cantide and these chemotherapeutic drugs, SAS software and Jin Zhengjun method were used. RESULTS: Combination treatments with 0.1μmol/L Cantide reduced the IC50 of DDP, 5-FU and ADM from 1.07, 4.15 and 0.29 μg/mL to 0.25,1.52 and 0.12 μg/mL respectively. The inhibition ability of DDP, 5-FU and ADM respectively in combination with Cantide in these tumor cells was higher than that of these drugs alone (P<0.0001). And synergism (Q≥1.15)was observed at the lower concentration of DDP (≤1μg/mL) and ADM (≤0.1 μg/mL) with combination of Cantide. In vivo, combination treatment with Cantide and DDP produced the greater growth inhibition of human liver carcinoma cells HepG2 in nude mice (0.65±0.19 g tumor) compared with that when only one of these drugs was used (Cantide group: 1.05±0.16 g tumor, P= 0.0009<0.001; DDP group: 1.13±0.09 g tumor, P= 0.0001<0.001). CONCLUSION: These findings indicate that Cantide may enhance therapeutic effectiveness of chemotherapeutic drugs over a wide range of tumor cells in vitro, and the combination use of Cantide and DDP can produce much higher inhibition rates, as compared with when either of these drugs was used only in vivo.