0.05),and T max had signif?icant differences(P

Objective To investigate the effects of miR-135a on HOXA10 expression,proliferation and apoptosis of SKOV3 cells.Methods (1) Through computer-aided algorithms,the predicted target gene of miR-135a (HOXA10)were determined.(2) miR-135a mimics,miR-135a inhibitor and negative control were transfected into SKOV3 cells,respectively.Reverse transcription (RT)-PCR,western blot analysis were used to examine the expression levels of HOXA10 at different times (24,48 and 72 hours).(3) A luciferase reporter assay was used to confirm the direct regulation between miR-135a and HOXA10.(4) SKOV3 cells proliferation at different times (24,48 and 72 hours) was detected by methyl thiazolyl tetrazolium(MTT) assay [quantified by absorbance(A)].Western blot was used to examine the expression of apoptosis-associated protein bcl-2,bax and caspase-3 in SKOV3 cells after 48 hours transfection.Results (1) HOXA10 was predicted to be the target gene of miR-135a by computer-aided algorithms.(2) RT-PCR shown that HOXA10 mRNA levels were decreased over time (24,48 and 72 hours) after miR-135a mimics transfectionin SKOV3 cells (0.94 ±0.04 vs 0.78 ±0.03 vs 0.70 ±0.03,P ＜0.05).While,the expression of HOXA10 mRNA was increased over time after miR-135a inhibitor transfection (1.14 ± 0.05 vs 1.16 ±0.03 vs 2.60 ±0.08,P ＜0.05).After transfected with miR-135a mimics or miR-135a inhibitor over 48 and 72 hours,the HOXA10 expression levels in SKOV3 cells were significantly lower or higher than each control group,respectively (all P ＜ 0.01).Western blot analysis of HOXA10 expression in SKOV3 cells confirmed the results of RT-PCR detected.(3) After cotransfection of miR-135a plasmid and pMIR-REPORT luciferase plasmid containing HOXA10,luciferase reporter assays showed that the luciferase activity reduced by 67.8％ (P ＜0.01).(4) MTT showed that SKOV3 cells growth after miR-135a mimics transfection for 48 and 72 hours were significantly lower than those in control group (0.38 ± 0.03 vs 0.52 ± 0.05,0.67 ±0.05 vs 0.75 ± 0.06 ; respectively,all P ＜ 0.05).While,SKOV3 cells transfected with miR-135a inhibitor for 72 hours grew significantly faster than that in control group (0.95 ± 0.05 vs 0.75 ± 0.06,P ＜ 0.01).After miR-135a mimics transfection,the level of bcl-2 protein was significantly lower than that in control group (0.28 ±0.06 vs 0.76 ±0.09,P ＜0.01).The activity of caspase-3 was significantly higher than that in control group (115.0 ± 2.4 vs 95.4 ± 2.1,P ＜ 0.01).While,there was no statistical difference of bax expression (P =0.142).However,after miR-135a inhibitor transfection,the expression level of bcl-2 protein was significantly higher than that in control group (0.92 ± 0.03 vs 0.76 ± 0.09,P =0.037) and the activity of caspase-3 was significantly lower than that in control group (59.5 ± 4.1 vs 95.4 ± 2.1,P ＜ 0.01).There was also no statistical difference of bax expression (P =0.066).Conclusion miR-135a may play an important role in cell proliferation and apoptosis of ovarian cancer cells by regulating HOXA10 and its downstream pathways.

OBJECTIVE: To investigate the effect of electro-acupuncture (EA) at Fenglong (GV 16) on body weight, blood lipids, nitric oxide (NO) and endothelin (ET) in rats with hyperlipidemia (HLP). METHODS: Eighty Wistar rats were randomly divided into normal control group (fed normal diet), untreated group (fed a high-fat diet), EA-treated group (fed a high-fat diet plus EA therapy) and pravastatin-treated group (fed a high-fat diet plus pravastatin tablet). There were 20 rats in each group. The body weight and the blood content of total cholesterol (TC), triacylglycerol (TG), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), NO the and ET of the rats in different groups were measured before experiment and after 30-day treatment. A modified method of cardiac puncture for blood sampling was used for blood collection. RESULTS: Compared with the normal control group, the body weight and the levels of TC, TG, LDL-C and ET in the untreated group were significantly elevated (P<0.01, P<0.05), while the levels of HDL-C and NO were obviously decreased (P<0.05). The body weight and the levels of TC, TG, LDL-C demonstrated significant reduction in pravastatin-treated group and EA-treated group as compared with the untreated group (P<0.01), and the NO content in pravastatin-treated group and EA-treated group was higher than that in the untreated group (P<0.01). Compared with the untreated group, HDL-C level was elevated significantly in pravastatin-treated group, while HDL-C level in EA-treated group was not changed significantly, and there was significant difference in the HDL-C level between pravastatin-treated group and EA-treated group (P<0.01). The level of ET was decreased obviously in pravastatin-treated group (P<0.05), while the level of ET in EA-treated group was not changed significantly (P>0.05). CONCLUSIONS: Both EA therapy and pravastatin have efficient regulation of body weight and the content of TC, TG, LDL-C and NO in HLP rats. To some extent, they are able to regulate the imbalance between ET and NO content under the condition of HLP. Western medicine such as pravastatin can regulate the HDL-C level in HLP rats, while the effect of EA therapy on regulation of the HDL-C level is limited.

Objective To study the effect of motor imagery therapy combined with conventional rehabilitation treatment on walking ability in patients with post-stroke hemiplegia.Methods Eighty patients with post-stroke hemiplegia were randomly divided into a treatment group (40 cases) and a control group (40 cases).All the patients in both groups were given basic medication and conventional rehabilitation treatment.In addition,the patients in treatment group were given motor imagery therapy.The 10 m maximum walking speed (10 m MWS),stride length,cadence,Fugl-Meyer motor assessment (lower limb) (FMA-L) and Holden's functional ambulation classification (FAC) were used to evaluate walking ability before the beginning of training and at the end of six weeks of training.Results Before intervention there was no significant difference between the two groups in terms of all the assessment (P ＞0.05).At the end of training,all measurements in both groups [the treatment group:10 m MWS (0.53 ± 0.20)m/s,stride length (78.91 ± 20.46) cm,cadence (78.10 ± 12.03) min,FMA-L (24.13 ±5.77),FAC (3.60±1.01);the control group:10 m M WS (0.42 ±0.15)m/s,stride length (69.75 ± 18.31)cm,cadence (71.14±9.29)/min,FMA-L (20.65 ±4.70),FAC (2.93 ±0.89)] were significantly better than those before training (P ＜ 0.05),and the improvements in treatment group were significantly better than those in control group (P ＜ 0.05).Conclusion Motor imagery therapy combined with conventional rehabilitation treatment can distinctly improve the walking ability in patients with post-stroke hemiplegia.

Objective To evaluate the efficacy and safety of laparoscopic technique in the treatment of refractory adhesive intestinal obstruction. Methods In a retrospective matched?pair analysis, 68 patients of re?fractory adhesive intestinal obstruction whose relapsed more than 3 times were included in the research. All patients were treated with laparoscopic operation or continued follow?up observation after conservative treatment from Janu?ary 2011 to January 2016. The patients were divided into 2 groups, laparoscopic surgery (32 cases) and observation group (36 cases), to contrast the recurrence rate and safety of the two types of treatment. Results The recurrence rate of laparoscopic surgery group was significantly lower in the observation group (18.8%vs. 77.8%, P<0.05). Re?fractory adhesive intestinal obstruction is further divided into Mixed type, Patchy adhesion type and Cable Belt Com?pression type, in the laparoscopic surgery subgroup analysis, hybrid recurrence rate (66.7%, 4/6 cases) was signifi?cantly higher than that of patchy adhesion type (10.0%, 1/10 cases) and cord compression type (6.3%, 1/16 cases). Conclusion Laparoscopic operation is safe and feasible in the treatment of refractory adhesive intestinal obstruc?tion, and then it can effectively reduce the recurrence rate of refractory adhesive intestinal obstruction;the curative effect is closely related to the type of adhesion.

Background and purpose:Liquid biopsy is a kind of blood, urine and other non-solid biologi-cal tissue sampling analysis, mainly for malignant tumor diagnosis, monitoring and predicting its prognosis. In this research, we optimized the extraction of miRNA in urine, established a standardized means of liquid biopsy, screened and verified the miRNA markers in patients with bladder cancer.Methods:From Jan. 2014 to Sept. 2015, we used miRNA microarray in six patients with bladder cancer and six healthy controls. Samples of 78 cases of bladder cancer and 23 healthy controls were tested by real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) to verify the relationship between miRNA markers in liquid biopsy and clinical pathological parameters. The diagnostic value of miRNA markers was also analyzed and compared.Results:We screened 10 miRNAs differential expression in urine. Combined with previous literature, we selected 20 miRNAs to verify their expression levels in bladder cancers and healthy controls. miR-509-5p/miR-124 ratio in the urine was found higher in patients with bladder cancer than in healthy controls (P<0.0001). With the rise of miR-509-5p/miR-124 ratio in urine, tumor stage and grade were also increased (P=0.003). When the cutoff was set at 0.41, the diagnostic sensitivity and specificity of miR-509-5p/miR-124 ratio were 73.1% and 82.6%, respectively. The AUC of miR-509-5p/miR-124 ratio to detect bladder cancer was 0.864, higher than that of urinary exfoliated cells (P=0.0002).Conclusion:We optimized the extraction of miRNAs in urine,established a standardized liquid biopsy of miRNA markers. The miR-509-5p/miR-124 ratio could be an ideal diagnos-tic marker for bladder cancer.

Objective To investigate the clinical features of gynecological malignant tumor related multiple primary malignant neoplasms (MPMN).Methods Apply retrospective and comprehensive analysis to the clinical data of 30 patients with gynecological malignant tumor related MPMN.Results Synchronous MPMN were found in 9 patients.Their average age was 50.2 years old and their median age was 49 years old.The neoplasms were located at ovary,uterus,cervix,breast and intestine.Metachronous MPMN were found in 21 patients.Their average age was 57.7 and their median age was 57 years old.The median interval between the first and the second primary malignant neoplasm was 4.0 years.The neoplasms were located at breast,ovary,uterus,gastrointestinal tract,uterine cervix,lung etc.In 30 cases,26 of them were treated by surgical operation and further adjunctive treatment of chemotherapy and (or) radiotherapy was conducted as per the neoplasm staging and its pathological results.The rest 4 patients (first primary malignant neoplasms were excised from 3 of them and another one was not treated by surgical operation) received adjunctive treatment of chemotherapy and (or) radiotherapy.Followed ups,which varied from 6 to 60 months,were made to 29 patients and 20 out of the 29 were alive.5-year survival rate of patients with gynecological malignant tumor related MPMN was 47.8％,2-year survival rate was 73.9％,and 1-year survival rate was 88.6％.Conclusion Pay more attention to the patients with gynecological malignant tumor related MPMN,examine the high-risk patients with malignant tumor comprehensively,identify whether it is recurrence,metastasis or new growth of malignant neoplasm,and further ensure early diagnosis and proper treatment,avoiding misdiagnosis and missed diagnosis.

Wistar rats were intragastrically administered with different doses (2, 5 and 10 g / kg body weight) of haematitum. 1H NMR-based metabonomic analysis coupled with multivariate statistical analysis (principal component analysis and partial least squares-discriminant analysis) was used to analyze the metabolic profiles of the urine samples collected from the treated rats. Univariate analysis on the 1H NMR spectra of urine (1 d before administration, 1-5 d post administration) was used to screen out the potential features of haematitum. Significant treatment related changes were observed for the levels of citrate, tuarine, creatinine,α-ketoglutarate, succinate and dimethylglycine, which could be used as potential features of haematitum. A trend of recovery in connection with dose levels was observed overtime. Such biochemical changes indicated that haematitum treatment at the dose of 2, 5 and 10 g / kg body weight affected the Krebs cycle and glucose metabolism, energy metabolism, choline metabolism and dimethylglycine metabolism in rats. These changes may attribute to the disturbances of hepatic function in 10 g / kg body weight group.

objective At present, most operable breast cancers are confirmed by excision biopsy during the operation. If the breast cancer was definitely diagnosed by Fine -Needle Aspiration cytology(FNA) preoperatively, the excision biopsy could be avoid. Methods We diagnosed 118 inpatients of breast mass with physical examination and FNA from September, 1995 to May, 1996. Results The sensitivity and positive predict value of FNA for diagnosing breast cancer are 95.7% and 97.1%. When the palpation and FNA are both positive, the doublepositive predict value for breast cancer is 100% in our study. It is the indication of using FNA that a breast mass was diagnosed as cancer by physical examination. We can treat a breast mass with redical operation directly without excision biopsy, if the breast mass was confirmed as breast cancer by both physical examination and FNA. Conclusion We concluded that the preoperative FNA could partially replace the excision biopsy for confirming breast cancer.

Objective Re stenosis is the common complication of stenting. This study was designed to investigate the functional changes of fibroblast in local re stenosis esophageal tissue after stenting and its relation to re stenosis. Methods Sixteen healthy adult dogs were divided into four groups. Esophageal stent was placed by means of “autogenous broad fascia transplantation and fixation”. At the end of 1,2,4 and 8 weeks,the dogs were killed, and the esophageal tissue with stent were taken out and analyzed by gross observation, light microscopy and electron microscopy. The expression of proliferation cell nuclear antigen (PCNA) and alpha smooth muscle actin (? SMA) in esophageal tissue at 1,2,4 and 8 weeks after stenting were studied by immunohistochemistry. The contents of hydroxyproline and gross amino acid(AA) in re stenosis tissue were measured by amino acid analysis. Results At week 1 and 2 the inflammatory reaction occurred evidently in stented esophagus, with granulation and fibrosis; in some places esophageal tissues began to proliferate towards the lumen. At week 1 some fibroblasts began to express PCNA and ? SMA, and at week 2 the expression augmented significantly. The content of hydroxyproline and AA was significantly higher in the esophageal tissues at 1 and 2 weeks after stenting than that in normal esophagus. At week 4 and 8 esophageal lumen became narrow conspicuously, with a lot of fibrotic tissue and few inflammatory cells. Only a few fibroblasts displayed the expression of PCNA instead of the expression of ? SMA at week 4. There was no expression of PCNA and ? SMA at week 8. The content of AA at week 4 increased significantly compared with that at week 2, and the level was similar between 4 and 8. Electron microscopy revealed that the fibroblasts were in the state of vigorous proliferation and secretion in esophageal tissue at week 2, and a lot of fibrotic tissue formed at week 8. Conclusions Re stenosis mainly expressed as granulation and fibrosis. At week 4 and 8 the fibrosis becomes stable gradually with the lessening of inflammatory reaction. Within 4 weeks of stenting fibroblast retains active proliferation and secretion, but after week 4 the function of fibroblasts gradually lessen or even lose.