Objective:To investigate the relationship of the mRNA expression of BRCA1 andβ-tubulinⅢwith docetaxel-resistance in esophageal cancer. Methods:The genes BRCA1 andβ-tubulinⅢwere determined at the mRNA level using RT-qPCR in 36 esophageal carcinoma specimens. Results:The mRNA expression of BRCA1 andβ-tubulinⅢwas determined in the 36 tumor samples using RT-qPCR. The median BRCA1 mRNA expression level in relation to that ofβ-actin was 6.27. The medianβ-tubulinⅢmRNA expression level in relation to that ofβ-actin was 4.44. The patients were divided into two groups using these cutoff values. The BRCA1 mRNA expression level was not correlated with the sensitivity of esophageal cancer patients to docetaxel (P=0.733). The response rate of the tumors with highβ-tubulinⅢexpression was (38.9%), which is significantly lower than in patients with lowβ-tubulinⅢexpression (83.3%) (P=0.015). Conclusion:Theβ-tubulinⅢexpression levels in the tumor tissues were probably an important biomarker for the efficacy of docetaxel chemotherapy in esophageal cancer patients. Our study may provide new insights into taxane chemotherapy for advanced esophageal cancer patients.

BACKGROUND:Stem celltherapy for renal ischemia-reperfusion injury has been the hot topics for many scholars. Its mechanism is very complex, which could not be explained by simple mechanism of stem cells differentiation. It is the result involving a variety of mechanisms. OBJECTIVE:To investigate the influence on immune cells during the bone marrow mesenchymal stem celltherapy for renal ischemia-reperfusion injury, then to preliminary summarize the immune regulation mechanism of bone marrow mesenchymal stem celltherapy for renal ischemia-reperfusion injury. METHODS:First, we established a model of renal ischemia-reperfusion injury in rats and, cultured and purified rat bone marrow mesenchymal stem cells in vitro. Then, the bone marrow mesenchymal stem cells were transplanted into the rat models. Using flow cytometry detection technology, we analyzed the proportion of CD4+CD25+regulatory T cells of rat spleen cells, discussed the effects on immune cells during the bone marrow mesenchymal stem celltherapy for renal ischemia-reperfusion injury, and then transferred the rat’s spleen cells to the nude mice which were subjected to renal renal ischemia-reperfusion injury. Renal function and renal histological changes of nude mice were assessed. RESULTS AND CONCLUSION:The bone marrow mesenchymal stem celltransplantation could significantly inhibit the decrease of CD4+CD25+regulatory T cellof spleen cells in rats with renal ischemia-reperfusion injury. The transplantation of spleen cells from the above-mentioned rats to nude mice could obviously protect nude mice from renal ischemia-reperfusion injury, characterized by lower serum creatinine, blood urea nitrogen and renal tubule pathologic damage score. Therefore, bone marrow mesenchymal stem cells have protective effects on renal ischemia-reperfusion injury by regulating the immune system.

Objective To investigate the relationship between interleukin (IL)-28B gene polymorphisms (rs12979860 and rs8099917) and treatment response in patients with chronic hepatitis C in China.Methods Taqman probes single nucleotide polymorphism genotyping methods were used to detect the genotypes of rs12979860 (C/T) and rs8099917 (T/G) located at IL-28B gene in 105 included patients.The patients were treated with standard doses of pegylated interferon plus ribavirin and were followed up regularly for therapeutic response and adverse reaction.The relationship between IL-28B gene polymorphism and antiviral treatment response of patients were analyzed.Categorical data were analyzed using Pearson chi-square test or Fisher exact test.Results Totally 105 cases were included in our study and 2 cases lost to follow-up because of moving away.Eight-one cases (78.6%) of the remaining 103 patients were CC/TT genotype (CC/TT group) at rs12979860 and rs8099917, 19 cases (18.4%) were CT/TG (CT/TG group) and 3 cases (2.9%)were TT/TG (TT/TG group).No other genotypes were detected and linkage disequilibrium was discovered at the two polymorphism loci (r2=0.11).After 4 weeks of treatment, 35 cases (43.2%) in CC/TT group, 3 cases (15.8%) in CT/TG group and non in TT/TG group achieved rapid virological response (RVR).There were statistically significant differences among three groups (P=0.033).After 12 weeks of treatment, 45 cases (55.6%) in CC/TT group, 6 cases (31.6%) in CT/TG group and none in TT/TG group achieved early virological response (EVR).There were statistically significant differences among three groups (P=0.025).At the end of the treatment, 68 cases (83.9%) in CC/TT group, 10 cases (52.6%) in CT/TG group and only 1 case (33.3%) in TT/TG group achieved end-of-treatment response (ETR).There were significant statistical differences among the three groups (P=0.003).After 24 weeks of follow-up, 62 cases (76.5%) in CC/TT group, 9 cases (47.4%) in CT/TG group and 1 case (33.3%) in TT/TG group achieved sustained virological response (SVR).There were statistically significant differences among the three groups (P=0.014).One hundred and one cases in CC/TT group developed adverse events, among them 19 cases needed clinical treatment.There were 43 cases in CT/TG group developed adverse events and 9 cases needed treatment.Seven cases in TT/TG group developed adverse events and only 1 case needed treatment.There were no statistically significant difference among three groups (χ2=0.139,P>0.05).Conclusions The genotype of rs12979860 (C/T) and rs8099917 (T/G) at IL-28B gene could affect the treatment response in patients with chronic hepatitis C.RVR and SVR are higher in patients with genotype CC/TT, which might help to guide HCV treatment.

Objective: To explore early diagnostic and resent prognostic assessment value of serum levels of ischemia-modified albumin (IMA) and cardiac troponin I (cTnI) for acute coronary syndrome (ACS).Methods: A total of 175 ACS patients were selected, including 73 cases with unstable angina pectoris (UAP), 61 cases with non-ST elevation myocardial infarction (NSTEMI) and 41 cases with ST elevation myocardial infarction (STEMI).According to chest pain-to-visit time, ACS patients were divided into <3h group (n=112) and 3~6h group (n=63);another 40 healthy subjects were selected simultaneously as healthy control group.Serum IMA and cTnI levels were compared among above groups and between patients suffering from major adverse cardiovascular events (MACE) within 30d or not in <3h group.Risk factors for MACE in <3h patients within 30d were screened.Results: Compared with healthy control group and UAP group, there were significant rise in serum levels of IMA[(16.78±4.25) μg/L, (35.16±8.32) μg/L vs.(49.76±9.29) μg/L, (52.07±11.34) μg/L], cTnI[(0.17±0.06) ng/ml, (0.15±0.06) ng/ml vs.(7.65±1.29) ng/ml, (8.83±1.40) ng/ml]in NSTEMI group and STEMI group, and IMA level in STEMI group was significantly higher than that of NSTEMI group, that of UAP group was significantly higher than that of healthy control group (P<0.05 or <0.01);serum IMA level of <3h group was significantly higher than those of 3~6h group and healthy control group, and cTnI level of <3h group was significantly lower than that of 3~6h group (P<0.01 all);serum levels of cTnI and IMA in patients suffering from MACE in <3h group were significantly higher than those of patients without MACE (P<0.01 both);multi-factor Logistic regression analysis indicated that elevated serum IMA level was an independent risk factor for MACE within 30d in ACS patients[OR=2.757,95%CI(2.084~4.705), P=0.001].Conclusion: The levels of cTnI and IMA significantly rise in ACS patients.IMA level possesses early diagnosis and recent prognosis evaluation value.

Objective:To study the biocompatibility of fibrin sealant (FS) and human corneal fibroblasts (HCFs) obtained by small incision lenticule extraction (SMILE).Methods:The human corneal stromal tissues were selected from corneal stromal lens in 24 eyes of 12 patients underwent SMILE in the First Affiliated Hospital of Guangxi Medical University from March to April 2018.HCFs were isolated and cultured in vitro within 1 hour after the corneal stromal lens were extracted and the growth status of HCFs on FS surface was observed.HCFs were divided into 2-fold leaching solution group and normal control group, and the cells in the two groups were treated with 2-fold leaching solution or complete medium according to grouping, respectively.The apoptosis of HCFs in the two groups was observed by acridine orange (AO)/ethidium bromide (EB) double staining.The proliferation of HCFs in the two groups was assayed by methyl thiazolyl tetrazolium (MTT) method.HCFs in logarithmic phase were divided into 2-fold leaching solution group, normal control group, and the cells were treated with 2-fold leaching solution or complete medium according to grouping, respectively.In addition, a blank control group without HCFs was also set and treated with complete medium.The absorbance value and relative growth rate of HCFs in the three groups were compared.HCFs in logarithmic phase were divided into 1-fold leaching solution group, 2-fold leaching solution group and normal control group, and the cells were treated with 1-fold leaching solution, 2-fold leaching solution or complete medium culture according to grouping, respectively.The apoptosis of HCFs in the three groups was compared by Annexin V-FITC/PI flow cytometry, and the cytotoxicity of the three groups was graded.Written informed consent was obtained from each patient before the operation.The study protocol adhered to the Declaration of Helsinki and was approved by the Ethics Committee of the First Affiliated Hospital of Guangxi Medical University (No.2018[022]). Results:HCFs grew well on FS surface and the morphology was normal.MTT assay showed that HCFs in the 2-fold leaching solution group and the normal control group had a similar proliferation tendency, and the toxicity index of HCFs in the 2-fold leaching solution group was graded 0-1 at 0-72 hours after changing solution.After AO/EB staining, the HCFs in the 2-fold leaching solution group and the normal control group were normal, and only a small amount of early apoptotic cells were observed.Flow cytometry showed that the apoptosis rates of the normal control group, once leaching solution group and the double leaching solution group were (4.96±1.09)%, (3.66±1.35)% and (2.88±0.66)%, respectively, with no significant difference among them ( F=2.89, P=0.13). Conclusions:FS has no cytotoxicity and has good biocompatibility with HCFs in vitro.

According to the theory of in vitro radioassay, using TSHAb as binder and 125I-staphylococcal protein A (125I-SPA) as tracer agent, a new method for detecting the abnormal immunoglobulin (aIgG) in the sera of patients with Graves disease (GD) was reported. In the initial study of the method, the most appropriate interacting conditions of TSHAb with aIgG were explored and compared with ELISA using ganglioside (GLS) as a binder. The relationship between aIgG and thyroid-stimulating imunoglobulin(TSI) was probed into. The results showed that TSHAb could interact specifically with aIgG in vitro; but it could not interact with IgG from sera of normal subjects, systemic lupus erythematosis (SLE) patients and diabetic patients. With aIgG as an evaluating index, the mean value in 29 normal subjects was 1. 06?0. 17. When the sera of 72 patients with GD in different clinical stages were studied, the aIgG index was positive in 83% of untreated GD patients (n = 24), in 12% of GD patients in clinical remission (n = 25) and in 82% of relapsed GD patients (n = 23). Very significant difference was observed between normal subjets and untreated and relapsed patients with GD (P

ObjectiveTo evaluate the clinical efficacy of bladder hydrodistention and intravesical sodium hyaluronate in the treatment of interstitial cystitis (IC).Methods Twenty-one IC patients received intravesical sodium hyaluronate therapy under combined blockage or intravenous anesthesia.Bladders were perfused with normal saline under 100 cm H2O perfusion pressure and expanded for 10 min,the bladders were then injected through a catheter with 40 mg/50 ml sodium hyaluronate which was released after 1 h.Intravesical perfusion was applied once every week four to six times in a course of treatment.Results The average bladder capacity was extended from 191.6 ± 88.7 ml before expansion to 425.3 ± 79.8 ml after bladder expansion ( P =0.000).The extension was done under anesthesia.There were two suspected bladder ruptures after starting the bladder expansion at 6.5 min and 7.2 min.There was significant gross hematuria in 19 cases,10 min after bladder expansion.After treatment,the catheters were removed 24 h after manipulation in 17 patients; the catheters were removed 72 h after manipulation in two cases with hematuria;the catheters were removed four days after manipulation in the two cases of suspected bladder rupture.Pain was significantly reduced after the catheters were removed and the maximum urinary output increased slightly.The day before the second injection of sodium hyaluronate,the urinary frequency decreased significantly than before start of treatment (32.8 vs 18.5 times/24 h).The maximum urinary output increased significantly compared with the output before treatment (86.7 vs 151.9 ml).Pain was reduced significantly after treatment (8.7 vs 3.0).The O'Leary-Sant IC score and the QOL were significantly improved (30.0 vs 17.0,5.9 vs 2.4,respectively) (P =0.000).After the third treatment,the symptoms continued to improve.The treatment results were best in the fifth week at the time of the sixth injection of sodium hyaluronate.Symptoms rebounded at six months.However compared with that before treatment,the difference was still statistically significant ( P =0.000).ConclusionsBladder hydrodistention under anesthesia for severe intractable IC patients produces immediate effectiveness.Sodium hyaluronic infusion can alleviate urinary frequency and pain,and the effectiveness and duration of treatment are positively correlated.

Objective To investigate the effects and possible mechanisms of platelet-activating factor (PAF) on the invasion of ovarian cancer cells and to provide a potential target for ovarian cancer therapy.Methods ( 1 ) Serous type ovarian cancer cell line OVCA429 with platelet-activating factor receptor (PAFR) positive and mucinous type cell line RMUG-L (PAFR negative) were treated with 100 nmol/L of the PAF,cell invasion ability was determined by transwell cell migration assay.(2) For determination of the optimal PAF concentration,ovarian cancer cell OVCA429 was treated by 0,0.1,1,10,100,and 1000 nmol/L of PAF for 10 minutes or 24 hour,respectively.To observe the different time point of protein changes,OVCA429 were treated by 100 nmol/L of PAF for 0,5 minutes,10 minutes,30 minutes,1 hour or 12 hours,respectively.The total proteins of treated cells were extracted according to standard protocol.The expression of p38 mitogen-activated protein kinase (p38 MAPK),phosphorylated p38 MAPK (p-p38 MAPK),transcription factor response element-binding protein (CREB),phosphorylated CREB (p-CREB) and matrix metalloproteinase-2 (MMP-2) were detected by western blot.(3) To verify the pathway involved in the PAF induction of the cancer cell invasion,we repeated the experiments by adding the inhibitors when treating cells with PAF.The inhibitors used were as follows,PAFR inhibitor-WEB2076 (50 μmol/L),pp38 MAPK inhibitor-SB203580 (10 μmol/L),CREB binding protein (CBP)-CREB interaction inhibitor217505(25 μ mol/L).All treated cells were divided into 6 groups:control group,PAF group,PAF + WEB2076 group,PAF + SB203580 group,PAF + 217505 group and PAF + SB203580 + 217505group.Results ( 1 ) By transwell assay,100 nmol/L of PAF could improve the invasion ability of OVCA429 cell significantly [ PAF:( 118 ± 23 ) cells vs.control:(36 ± 8 ) cells,P ＜ 0.0l ],while the same treatment had no effect on RMUG-L cells [PAF:(45 t 13) cells vs.control:(53 ±9) cells,P＞0.05].(2) Even a very low concentration of PAF (0.1 nmol/L) could increase the expression of p-CREB and MMP-2,while the most effective concentration of PAF was 100 nmol/L.The highest p-CREB protein expression was detected at 10 minutes after administration of 100 nmol/L PAF,as well as the expression of p-p38 MAPK protein.Even 12 hours after treatment the p-p38 MAPK protein could be detected,while there was no significant difference in the expression of CREB ( P ＞ 0.05 ).(3) As compared with PAF group,both in PAF + WEB2076 group and PAF + SB203580 group,the expressions of p-p38 MAPK,p-CREB and MMP-2 protein were decreased significantly; in PAF + 217505 group,although the expression of p-p38 MAPK and p-CREB protein was significantly higher than the control group,the expression of MMP-2 protein was significantly lower; in PAF + SB203580 + 217505 group,the expression of these three proteins were also significantly lower,but there was no significant difference as compared with that in the PAF + WEB2076 or PAF + SB203580 group.Conclusion PAF could induce MMP-2 expression and contributed to PAFR positive ovarian cancer cell invasion via activation of CREB by phosphorylating of p38 MAPK.

Objective To explore genes expression of adiponectin receptors during differentiation of SW872 preadipocytes. Met-(hods) SW872 preadipocytes were cultured in vitro and induced to differentiate by 0.6 mmol/L oleic acid. During the progress of diffe-(rentiation), the morphological changes of SW872 cells were observed and the differentiation rate was assayed by oil-red O staining. Adiponectin receptors mRNA was measured by reverse transcription-polymerase chain reaction during differentiation of SW872 preadipocytes. Results 1.After stimulated by 0.6 mmol/L oleic acid for 72 hours, almost all SW872 cells were differentiated,and there were lots of fat droplets in the cells.2.There were adiponectin receptors genes expressions in SW872 preadipocytes.After 72 hours,and the levels of adiponectin receptor(AdipoR) 1 mRNA and AdipoR 2 mRNA were markedly increased up to 2.54 and 4.09 times,respectively. Conclusion There are AdipoR1 and AdipoR2 genes expressions in fat cells and the expressions are differentiation-dependent.

Objective To study the effect of oleic acid on the differentiation of SW872 preadipocytes.Methods SW872 prea-(dipocytes) were cultured and induced to differentiate by 0.6 mmol/L oleic acid in vitro.After 24 h,48 h and 72 h of differentiation,the morphological changes of SW872 preadipocytes were observed and the differentiation rate was assayed by oil-red O staining.In addition,triglyceride(TG) mass was detected by chemical colorimetry methods.During the differentiation of SW872 preadipocytes,transcription factors including peroxisome proliferator activated receptor-?_2(PPAR-?_2) and CAAT/enhancer binding protein-?(C/EBP-?)(mRNA) were also measured by reverse transcription-polymerase chain reaction(RT-PCR).Results 1.SW872 preadipocytes were fibroblastic and had no obvious fat droplet in cytoplasm.However,when stimulated for 72 hby 0.6 mmol/L oleic acid,SW872 preadipocytes became more bigger and rounder and differentiated into mature adipocytes with lots of fat droplets in the cells.2.Compared with that of predifferentiation,the concentration of TG mass increased by 14 folds after 72-hour differentiation(P