Objective To explore the method of culture of rat adipose-derived stem cells(ADSCs) and differentiation induction into myoblasts. Methods Adipose tissues were obtained from SD rats,and were isolated by enzyme digestion and cultured into ADSCs.The expression of surface antigen CD90,CD105 and CD34 was detected by immunofluorescence and flow cytometry.ADSCs of the second passage with logarithmic growth were obtained,and culture media containing 5-azacytidine(5-aza) and basic culture media were employed for cells in induction group and control group,respectively.The induction lasted for 7 d,14 d,21 d,28 d and 35 d,respectively.Cell growth and cell morphology were observed by inverted phase contrast microscope,and immunofluorescence and flow cytometry were utilized to detect the expression of myoblast specific antigens desmin and myosin. Results ADSCs were successfully isolated and cultured,and were identified to be stem cells.On the 28th day of induction,cells in induction group displayed "swirl" morpholgy,and multinucleation was observed.It was revealed by immunofluorescence and flow cytometry that the highest expression rates of desmin and myosin were 52.57% and 50.04%,respectively on the 28th day of induction,while there was no expression before induction and in control group. Conclusion ADSCs can be isolated and cultured from rat adipose tissues,and can further differentiate into myoblasts after induction by culture media with 5-aza.The expression of myoblast specific antigen is the highest on the 28th day of induction.

Objective:To investigate the clinical efficacy of gabapentin capsules( GBP)combined with amitriptyline and tramadol in the treatment of patients with postherpetic neuralgia( PHü)and anxiety. Methods:Totally 106 PHü patients with anxiety symptoms at different degree were selected and divided into group A(n=53)and group B(n=53). Group A was given GBP,amitriptyline and tramadol,while group was given GBP only. The anxiety,depression,quality of life and pain-relieving intensity( by VAS)of the pa-tients were determined before the medication( T1 )and in the first week( T2 )and the fourth week( T3 )after the medication. Follow-up was carried out regularly according to the requirements to evaluate the clinical efficacy. Results:The anxiety,depression and VAS scores in the two groups on T2 were statistically significant lower than the corresponding results on T1(P<0.05). On T3,however, there were statistically significant differences between the two groups(P<0. 05). The total adverse reaction rate of group A was signif-icantly lower than that of group B(P<0. 05). The total effective rate of group A was significantly higher than that of group B(P<0. 05). Conclusion:GBP combined with tramadol and amitriptyline in the treatment of PHü patients with anxiety can effectively con-trol the pain of the patients,significantly reduce the scores of VAS and improve the quality of life of the patients.

Long noncoding RNA (lncRNA) is a class of RNA molecules that longer than 200 nucleotides.lncRNA is lack of functional open reading frames and has no protein coding ability.Recently, accumulating evidences indicate that lncRNA plays active roles in tumor carcinogenesis and progression.The aberrant expression of lncRNA is significantly correlated with the growth,metastasis,and therapy-resistance of digestive system tumors.Thus,lncRNA may be served as new targets for the diagnosis,treatment,and progno-sis of patients with digestive system tumors.

Objective To study the expression of interleukin-1β in aortic dissections and aneurysms. Methods Aortic specimens were obtained from patients with type Ⅰ thoracic aortic dissection (11 cases),ascending thoracic aortic aneurysms (10 cases),and healthy organ donors (7 cases).Expression of interleukin-1β,matrix metalloproteinase-9,and signal transduction factors phospho-p38 and phospho-JNK were detected by real time RT-PCR,Western blot,and immunohistochemistry,respectively.TUNEL staining was performed to detect apoptosis of media cells. Results Apoptosis in the media of thoracic aortic dissection and ascending thoracic aortic aneurysms was dramatically higher than control group.Expression of interleukin-1β gradually increased in an order of control group,thoracic aortic dissection to ascending thoracic aortic aneurysms ( P ＜ 0.01,respectively).Expression of matrix metalloproteinase-9significantly increased in the media of thoracic aortic dissection and ascending thoracic aortic aneurysms compared with control group (P ＜ 0.01,respectively).There were positive correlations between interleukin1 β and matrix metalloproteinase-9,interleukin-1β and phospho-p38 in thoracic aortic dissection ( P ＜ 0.01,respectively),interleukin-1β and apoptosis in ascending thoracic aortic aneurysms (P ＜ 0.01 ).Conclusions Interleukin-1β and interferon-γ might effect the formation of thoracic aortic dissection and ascending thoracic aortic aneurysms possibly through the up-regulation of matrix metalloproteinase-9 and apoptosis of media cells in humans.

Objective: To observe the clinical efficacy of deep electroacupuncture (EA) at Baliao points in treating stress urinary incontinence (SUI). Methods: A total of 60 female patients with SUI were divided into two groups according to the order of consultation, with 30 cases in each group. The control group was treated with pelvic floor muscle training. The treatment group was treated with deep EA at Baliao points [Shangliao (BL 31), Ciliao (BL 32), Zhongliao (BL 33) and Xialiao (BL 34)]. Results: The total effective rate was 93.3％ in the treatment group, versus 33.3％ in the control group, and the total effective rate of the treatment group was significantly higher than that of the control group (P<0.05). After treatment, the scores of international consultation on incontinence questionnaire-short form (ICIQ-SF) and the volume of urinary leakage in both groups were lower than those before treatment (all P<0.05), and the ICIQ-SF score and the volume of urinary leakage in the treatment group were lower than those in the control group (both P<0.05). Conclusion: Deep EA at Baliao points with long needles can improve the clinical symptoms in female patients with SUI, and it has a better curative effect than pelvic floor muscle training.

Objective To investigate the expression levels of serum LINC00978 in gastric cancer patients and its correlation with clinicopathological characteristics,and evaluate its clinical diagnostic value.Methods The expression levels of LINC00978 in gastric cancer cells and serum samples from the patients with gastric cancer or gastric benign diseases and healthy controls were detected by real-time qRT-PCR,and its correlations with clinicopathological parameters were analyzed.The diagnostic efficacy of serum LINC00978 was evaluated by the receiver operating characteristic (ROC) curve.Results The expression levels of LINC00978 in MGC-803 cells were significantly higher than that in GES-1 cells (18.88 ± 1.15 vs 1.00 ± 0.03,t =-21.926,P ＜ 0.05).The expression levels (median[P25,P75]) of serum LINC00978 in gastric cancer patients (3.525 [1.385,8.954]) were significantly higher than those in the patients with gastric benign diseases (0.419 [0.258,1.369],Z =-4.834,P ＜ 0.01) and healthy controls (0.814 [0.351,2.510],Z =-4.686,P ＜ 0.01).The expression levels of serum LINC00978 in gastric cancer patients were significantly related to lymphatic metastasis (r =0.448,P ＜ 0.01),invasive depth (r =0.369,P ＜ 0.01) and TNM stage (r =0.383,P ＜ 0.01),and those after operation significantly lower than before operation (0.17 [0.15,0.39] vs 0.98 [0.59,1.61],Z =-5.731,P ＜ 0.01).There was no significant difference in serum LINC00978 levels between the patients with gastric benign diseases and healthy controls (Z =-1.693,P ＞ 0.05).The area under the ROC curve (AUCROC) and 95％ confidence interval (CI) of serum LINC00978 levels in the diagnosis of gastric cancer were 0.807 and 0.732-0.882,respectively.When the cutoff value was 1.806,the sensitivity and specificity for the diagnosis of gastric cancer were 71.4％ and 75.9％,respectively.Conclusion The expression levels of serum LINC00978 in gastric cancer patients increase and are related to lymphatic metastasis,invasive depth and TNM stage,which may be served as a novel biomarker for the diagnosis of gastric cancer.

Objective To investigate the application of fetuses with talipes equinovarus (TE) using chromosomal microarray analysis (CMA) technology. Methods From May 2012 to June 2015, 54 fetuses were found with TE and with or without other structural anomalies by prenatal ultrasound. Karyotyping was taking for them all, and the fetuses with normal karyotypes took another CMA test. The data were analyzed with CHAS software. Finally all the cases were followed up to know about their pregnancy outcomes. Results One of the 54 cases was detected with abnormal karyotype which was trisomy 18 (2%, 1/54). CMA was undertaken to the remaining fetuses, they were divided into 2 groups, including isolated TE group (n=38) and complex TE group (n=15). The detection rate of clinical significant copy number variations (CNV) by CMA was 11% (6/53), while isolated and complex TE group were 5% (2/38) and 4/15, respectively (P=0.047). Of the 53 cases, 51 cases were successfully followed up. Eleven cases were found without TE after birth, and the false positive rate (FPR) of TE was 22%(11/51). Conclusions Whole-genome high-resolution CMA increased the detection rate by 11% in fetuses with TE. With the FPR and the detection rate of the clinical significant CNV of 2 groups, whole-genome CMA could be recommended to the fetuses with complex TE group but normal karyotypes. A series of ultrasonic tests should be suggested to the isolate TE group, while with the abnormal ultrasound, fetuses would be suggested to have CMA test for decreasing the rates of invasive prenatal diagnosis and FPR.

Objective To investigate the effects of chronic hyperglycemia on the proliferation,secretion function and expression of endoplasmic reticulum stress(ERS) related molecules in pancreatic ? cell line MIN6. Methods MIN6 cells were treated with different concentrations of glucose(5.6,25 and 33.3mmol/L) and were harvested at indicated time(24h and 96h) for examinations.Cell proliferation was tested using CCK-8 solution,insulin and proinsulin secretion function was determined by ELISA,and mRNA expression of ERS related molecules(Total XBP-1,Spliced XBP-1) and protein expression of phosphorylated IRE1? were detected by Real-time PCR and Western blotting,respectively. Results After treatment with hyperglycemia for 96h, cell proliferation was significantly lower than that treated for 24h(P

Objective To evaluate the inter-fraction setup error during the treatment with megavoltage computed tomography (MVCT) and provide theoretical basis for clinical target volume-planning target volume (CTV-PTV) margins for nasopharyngeal carcinoma (NPC) patients treated with tomotherapy.Methods Thirty-seven consecutive NPC patients treated with tomotherapy were prospectively enrolled for the study between February 2015 and September 2015.For each patient,one MVCT scan was obtained after conventional positioning,online correction and tomotherapy delivery daily,and the scan was registered to the planning CT to determine inter-fraction setup error.The expanding margin for PTV (MPTV) was calculated with the recipe:MPTV =2.5∑ + 0.76 (∑:systematic error;6:random error).Results The average absolute errors of the inter-fraction were (2.102 ± 0.040 6) mm,(1.490 ± 0.034 8) mm,(1.306 ± 0.335) mm and (1.392 ± 0.038 4) ° in the three dimensions.Gradual increases in both inter-fraction three-dimensional displacement were observed with time and treatment (P ＜ 0.05).The total MPTV ac counting for inter-error were 3.467 5 mm,2.979 5 mm and 2.888 5 mm.Conclusions Tomotherapy irradiation technology personalized MPTV should be adopted for the design of tomotherapy plan.Displacement increased as a function of time.

MP-Rep fusion gene which size is about 1000bp was constructed by recombinant PCR technique.In order to construct fusion gene,Tobacco mosaic virus(TMV) partial movement protein gene(MP) and Cucumber mosaic virus(CMV) partial replicase gene(rep) were ligated.Two copies of MP-Rep fusion gene were ligated with soybean intron in inverted repeat manner,the recombinant fragments were then inserted into binary vector pBIN438 under the control of 35S promoter.Recombinant clone pBIN438-MP-Rep(i/r) which contained two different virus derived genes was constructed.Recombinant clone pBIN438-MP-Rep(i/r) in accord with expected design was certified by restriction endonuclease enzymes digestion and PCR analysis.This approach provides a basis for Broad-spectrum plant virus resistance mediated by RNA Silencing.