颅脑手术患者废弃脑组织中神经干细胞的培养与鉴定

目的:研究颅脑手术患者废弃脑组织中神经干细胞的培养与鉴定,为立体定向自体神经干细胞移植治疗脑出血、脑梗塞及颅脑损伤后遗症提供临床前基础。方法:通过改进原代培养方法,从颅脑手术患者废弃脑组织中培养和鉴定神经干细胞(NSCs)。收集颅脑手术患者不同脑区废弃脑组织各约500 mg以上,按改良方法进行原代培养并传代,通过免疫荧光检测NSCs标志物巢蛋白(Nestin)的表达,传代培养5代时开始进行诱导分化。结果:培养约3～10 d后,多数患者废弃脑组织中均可见干细胞球生长,经Nestin免疫荧光检测均呈阳性表达。选取第6代诱导分化后的贴壁细胞分别经β-Tubulin(神经元标志物)、Sox10(少突胶质细胞标志物)与GFAP(星形胶质细胞标志物)免疫荧光检测,可见少量神经元、少突胶质细胞及星形胶质细胞阳性表达。取第3代与第6代细胞行免疫印迹鉴定,第3代细胞仅见Nestin与少量β-Tubulin表达,第6代细胞可见不同程度的Nestin、β-Tubulin、Sox10及GFAP等表达。结论:本研究成功从颅脑手术患者不同脑区废弃脑组织中分离培养获得成人NSCs,并可向神经元及神经胶质细胞分化,使立体定向自体神经干细胞移植促进神经功能修复从实验室到临床应用成为可能。     

Lactulose Mediates Suppression of Dextran Sodium Sulfate-Induced Colon Inflammation by Increasing Hydrogen Production

Background

Molecular hydrogen (H₂) is a potent antioxidant and able to protect organs from oxidative stress injuries. Orally administered lactulose, a potent H₂ inducer, is digested by colon microflora and significantly increases H₂ production, indicating its potential anti-inflammatory action.

Objective

To evaluate the anti-inflammatory effects of lactulose on dextran sodium sulfate (DSS)-induced colitis in mice.

Methods

Mice were randomly assigned into seven groups, receiving regular distilled water, H₂-rich saline (peritoneal injection), DSS, oral lactulose (0.1, 0.15, 0.2 ml/10 g, respectively), and lactulose (0.2 ml/10 g) + oral antibiotics. The mouse model of human ulcerative colitis was established by supplying mice with water containing DSS. The H₂ breath test was used to determine the exhaled H₂ concentration. Body weight, colitis score, colon length, pathological features and tumor necrosis factor alpha (TNF-α), interleukin-1β (IL-1β), maleic dialdehyde (MDA) and marrow peroxidase (MPO) levels in colon lesions were evaluated.

Results

After 7 days, DSS-induced loss of body weight, increase of colitis score, shortening of colon length, pathological changes and elevated levels of TNF-α, IL-1β, MDA, and MPO in colon lesions, were significantly suppressed by oral lactulose administration and intraperitoneally injected H₂-rich saline. Ingestion of antibiotics significantly compromised the anti-inflammatory effects of lactulose. The H₂ breath test showed that lactulose administration significantly induced hydrogen production and that antibiotics administration could inhibit H₂ production.

Conclusion

Lactulose can prevent the development of DSS-induced colitis and alleviate oxidative stress in the colon, as measured by MDA and MPO, probably by increasing endogenous H₂ production.     

BMP7 Expression in Esophageal Squamous Cell Carcinoma and Its Potential Role in Modulating Metastasis

Background

Our previous study showed that BMP7 revealed significantly higher levels in esophageal squamous cell carcinoma (ESCC) tissues with lymph node metastasis compared with non-lymph node metastasis, using gene expression profiling assays. The roles of BMP7 in ESCC is not fully understood.

Aim

The aim of this study was to investigate the effect of BMP7 on lymph node metastasis of ESCC and to explore its potential mechanism.

Methods

Expression of BMP7 in ESCC tissues was evaluated by immunohistochemistry. BMP7 were down-regulated by RNA interference. The protein and mRNA levels of BMP7 were detected by western blot and RT-PCR, respectively. High content screening and transwell assay were used to identify the metastatic ability of tumor cells.

Results

Positivity of BMP7 staining was 57.5 % in the tissues of primary carcinoma with lymph node metastasis compared to tissues without lymph node metastasis, and expression of BMP7 was significantly higher in the cell lines with highly metastatic capacity than that in the cell lines without metastatic ability. Suppression of endogenous BMP7 expression by siRNA in the highly metastatic cell lines resulted in significant reduction in ability of cell migration and invasion in both in vitro and in vivo studies. In addition, inhibition of BMP7 by siRNA also leads to up-regulation of E-cadherin and down-regulation of MMP-9 in the highly metastatic cell lines.

Conclusions

These findings indicate that BMP7 modulates the expression of E-cadherin and MMP-9, and by which mechanism it may regulate cell migration and metastasis of ESCCs.     

改性甲壳素生物修复膜治疗浅Ⅱ度烧伤的临床疗效

目的探讨改性甲壳素生物修复膜治疗浅Ⅱ度烧伤的临床疗效。 方法将石家庄市第一医院烧伤整形科2016年5月至2018年5月收治的浅Ⅱ度烧伤患者70例按随机数字表法分为治疗组和对照组,每组35例。治疗组患者创面经过碘伏消毒后,外用0.9%氯化钠溶液冲洗,清除坏死腐皮及异物,外用改性甲壳素生物修复膜覆盖,每日换药1次,直至创面愈合;对照组同样清理创面后,外用无菌油纱覆盖创面,每日换药1次。观察伤后7、10、13 d创面愈合率及创面平均愈合时间、患者换药疼痛数字评分法(NRS)评分、创面感染情况、瘢痕情况及其他不良反应。数据比较采用t检验。 结果治疗组患者在伤后7、10、13 d创面愈合率分别为(83.66±3.59)%、(93.69±3.24)%、(99.46±0.78)%明显高于对照组(81.40±3.50)%、(90.63±4.25)%、(98.57±1.63)%,差异均有统计学意义(t＝2.66、3.38、2.90,P＝0.010、0.001、0.006);治疗组创面平均愈合时间为(12.11±1.89) d,短于对照组(13.51±1.15) d,差异有统计学意义(t＝3.75,P<0.05);治疗组患者NRS评分为(3.29±0.52)分,低于对照组(3.86±0.49)分,差异有统计学意义(t＝4.72,P<0.05),2组均未出现感染患者,随访3个月均未发现瘢痕生长,未发现其他不良反应。 结论应用改性甲壳素生物修复膜可提高创面愈合率,缩短浅Ⅱ度烧伤患者创面愈合时间,减轻患者疼痛。     

LINC00205 promotes proliferation,migration and invasion of HCC cells by targeting miR-122-5p

Long non-coding RNAs (lncRNAs) have been identified as crucial regulators in the tumorigenesis and progression of hepatocellular carcinoma (HCC). Recently, long intergenic non-protein coding RNA 205 (LINC00205) has been identified as a prognostic biomarker in HCC. However, the biological role of LINC0205 and its potential molecular mechanism are poorly investigated. Here, we found that the expression of LINC00205 was dramatically up-regulated in HCC tissues compared to adjacent nontumor tissues. Furthermore, the level of LINC00205 in both Hep3B and Huh7 cells was prominently higher than that in normal hepatic cell line LO2. Notably, the high expression of LINC00205 was strongly correlated with tumor size ≥5 cm, venous infiltration and advanced tumor stages. Functionally, LINC00205 knockdown obviously repressed the proliferation, migration and invasion of Hep3B and Huh7 cells in vitro. An inverse correlation between LINC00205 and miR-122-5p was detected in HCC tissues. Interestingly, LINC00205 knockdown increased the level of miR-122-5p in both Hep3B and Huh7 cells. Mechanistically, luciferase reporter assay demonstrated LINC00205 acted as a competing endogenous RNA (ceRNA) by directly interacting with miR-122-5p. More importantly, miR-122-5p overexpression significantly restrained the proliferation, migration and invasion of HCC cells. Collectively, our study provides solid evidence to support the oncogenic role of LINC00205 in HCC, which may be benefit for the improvement of HCC therapy.     

Genetic variants of DNA repair pathway genes on lung cancer risk

As is commonly perceived, polymorphisms in genes of deoxyribonucleic acid (DNA) repair pathway plays a fundamental role in defective DNA repair and mutagenesis prevention and serves to contribute to the individual susceptibility to the development of a variety of cancers. Recently, an increasing number of studies have been dedicated to the contentious and ambiguous links between polymorphisms in genes of DNA repair pathway and lung cancer (LC) risk. In response, a comprehensive updated meta-analysis has been proposed herein to assess the correlation between polymorphisms of DNA repair pathway genes and susceptibility to LC. This paper has identified and retrieved eligible articles from PubMed, Google Scholar, Web of Science, and CNKI databases till February 20, 2019. Finally, 295 case-control studies as to the fourteen polymorphisms of DNA repair pathway genes were enrolled. When the results have been pooled, we have brought to light the conclusion that ERCC2-rs13181 polymorphism has an elevated association with LC risk under allele, heterozygote, and dominant comparisons. In the subgroup analysis by ethnicity, we have found that the Caucasian individuals with “B” variant possess risk of LC which was more than twice as much as allele, homozygote, and recessive models. In comparison, Asian carriers of rs13181 polymorphism in ERCC2 gene are more susceptible to LC in heterozygote, dominant models. To sum up, ERCC2-rs13181 polymorphism could be a critical factor in stimulating LC evolvement. Future studies with a larger sample size and multivariate factors are needed to vindicate these findings.     

Expression and prognostic value of FOXP1 in esophageal squamous cell carcinoma

BackgroundForkhead box protein P1 (FOXP1) has been suggested as a prognostic marker in several malignant tumors. However, the significance of FOXP1 in esophageal squamous cell carcinoma (ESCC) is still unclear. The purpose of this study was to investigate the expression pattern of FOXP1 in normal esophageal tissue and ESCC and to analyze the clinicopathological significance and prognostic value of FOXP1 in ESCC.MethodsFOXP1 was detected by immunohistochemistry using tissue microarrays containing tumor tissues and adjacent normal tissues from 270 ESCC patients with oncological follow-up data.ResultsNormal esophageal tissues predominantly showed an exclusive nuclear FOXP1 (n-FOXP1) expression pattern, and no exclusive cytoplasmic FOXP1 (c-FOXP1) staining was found. In ESCC, the expression rates of exclusive n-FOXP1-positive, exclusive c-FOXP1-positive, both nuclear and cytoplasmic positive and complete negative were 14.4%, 28.9%, 10.4% and 46.3%, respectively. High n-FOXP1 expression was significantly correlated with decreased postoperative recurrence and distant metastasis (P < 0.05). Furthermore, elevated c-FOXP1 expression was significantly associated with regional lymph node metastasis and distant metastasis (P < 0.05). High c-FOXP1 expression had an effect on shorter overall survival (OS) time, but the difference was not statistically significant (P > 0.05). Kaplan–Meier analysis showed that ESCC patients with high n-FOXP1 expression survived significantly longer than patients with low n-FOXP1 expression. Multivariate analysis confirmed that patients with high n-FOXP1 staining exhibit good prognosis and n-FOXP1 was an independent factor for ESCC prognosis.ConclusionsOur results suggest that FOXP1 plays an essential role in ESCC progression and prognosis and may be a useful biomarker for predicting survival.     

MicroRNA-301b-3p accelerates the growth of gastric cancer cells by targeting zinc finger and BTB domain containing 4

MicroRNAs (miRNAs) have been found to be aberrantly expressed and exert essential roles in the tumorigenesis and progression of gastric cancer (GC). miR-301b-3p has been recognized as a cancer-related miRNA in lung cancer, bladder cancer and hepatocellular carcinoma. However, the function of miR-301b-3p in GC progression and its underlying mechanism have not been studied yet. In this study, we found that miR-301b-3p expression was up-regulated in GC tissues compared to adjacent noncancerous tissues. Furthermore, the elevated levels of miR-301b-3p were detected in GC cell lines (SGC-7901, AGS, MKN-45 and MGC-803) as compared with GES-1 cells. Interestingly, GC tissues from patients with tumor size ≥ 5 cm and advanced tumor stages showed obvious higher levels of miR-301b-3p compared to matched controls. Functionally, miR-301b-3p knockdown prominently inhibited cell proliferation, and induced cell cycle arrest at G1 phase and apoptosis in MGC-803 cells. Meanwhile, ectopic expression of miR-301b-3p conversely regulated these biological behaviors of MKN-45 cells. Next, we found that miR-301b-3p knockdown increased, whereas miR-301b-3p overexpression reduced the expression of zinc finger and BTB domain containing 4 (ZBTB4) in GC cells. Accordingly, luciferase reporter assay identified ZBTB4 as a direct target of miR-301b-3p. ZBTB4 overexpression markedly restrained the growth of MGC-803 cells. More importantly, ZBTB4 silencing partially reversed miR-301b-3p knockdown-induced tumor suppressive effects on MGC-803 cells. In conclusion, we firstly revealed that miR-301-3p was highly expressed in GC and contributed to tumor progression via attenuating ZBTB4, which might provide a novel molecular-targeted strategy for GC treatment.     

Referencing the trochlear groove based on three-dimensional computed tomography imaging improves the reliability of the measurement of the tibial tuberosity–trochlear groove distance in patients with higher grades of trochlea dysplasia

BackgroundTo determine whether 3D-CT imaging technique is valid and reproducible compared to conventional CT measurement technique (CCT) for the detection of a femoropatellar instability.MethodsPatients who had undergone surgery for femoropatellar instability (patellar instability group) between 2010 and 2016 (n = 37 knees of 35 patients) were retrospectively enrolled. For the matched control group, patients who had acute anterior cruciate ligament injury (< 4 weeks previously; n = 30) were recruited. Preoperative CT data had been obtained in all patients. Inter-rater reliability was calculated for both measurement protocols, and inter-method reliability was calculated between the two imaging modalities. The results are reported using intraclass correlation coefficients (ICCs) and Bland–Altman 95% limits of agreement.ResultsAll patients in the patellar instability group had femoral trochlear dysplasia (Dejour types A: four, B: 19, C: seven, and D: six), but no dysplasia was noted in the control group. In the patellar instability group, the CCT technique showed a poor inter-rater agreement (ICC = 0.74), and the 3D-CT technique still showed excellent inter-rater agreement (ICCs = 0.91). In the sub-analysis of the patellar instability group according to the trochlear dysplasia grade, ICCs were markedly decreased with severe trochlear dysplasia when using CCT technique; however, the 3D-CT technique could provide excellent reliability even with severe trochlear dysplasia.ConclusionThe 3D-CT imaging technique for the measurement of the TT–TG distance can be suggested as a better measurement technique for patellar instability patients with bone abnormality.