ObjectiveTo evaluate a new coloanal anastomosis preserving dentate line and anal sphincter. Methods After total mesorectal excision in 87 patients with low rectal carcinoma, the rectum no more than 1cm above the dentate line was preserved. The rectal mucosa was stripped and the dentate line was saved, then a sustaining anastomotic tube was fixed into the proximal colon, and the colon was pulled down and anastomosed with the remnant rectum 0.5cm above the dentate line. Results The ultralow coloanal anastomosis with anal sphincter preservation was accomplished. No perioperative death and anastomotic leakage occurred. The patients were followed up for 2 to 6 months and the follow-up rate was 89%. There was no anastomotic recurrence. Soft tissue recurrence in pelvic cavity was found in 3 cases, lymph node recurrence in obturator space recurrence in 2 cases and liver metastasis in 6 cases. Anastomotic stenosis was found in 6 cases 12 months later. The defecation function returned to normal six months after operation. Conclusions The sustaining banding method in the ultralow coloanal anastomosis with anal sphincter preservation is a safe and reliable surgical procedure.

Objective To evaluate a surgical procedure of low/ultralow colo-rectal(anal) anastomoses with sustaining bonding method after total mesorectal excision (TME) for lower rectal cancer. Methods After TME in 346 cases of lower rectal carcinoma, a sustaining anastomotic tube was inserted into the proximal colon, then the remnant was ligated and sutured. The rectal remnant no less than 1cm was preserved by colo-rectal anastomoses of modified Welch operation,while the rectal remnant no more than 1cm were preserved by colo-anal anastomoses with anal sphincter preservation. Results There was no perioperative mortality. Anastomotic leakage developed in 4 cases (1.2%), and anastomotic stenosis in 10 (2.9%). Postoperative 5 year survival and recurrence was 78.6%, 6.3% respectively. The defecation function was satisfactory in 82.6% cases. Conclusions Low/ultra-low colo-rectal(anal) anastomoses with sustaining bonding method after TME is safe and effective for lower rectal cancer.

Objective To study the influence of clinicopathologic characteristics and surgical treatment of gastric cancer on patients' survival rate.Methods From Apr.1994 to Aug.2005, the data of 759 gastric cancer patients concerning surgical treatment, pathological diagnosis and outcome were collected. Retrospective analysis of the results was made, 3-year and 5-year survival rates were calculated by Kaplan-Meier curve method, univariate analysis was done through Log-rank and multiple factors comparison through Cox regression analysis, and follow-up duration was 4-131 months.Results Single factor analysis indicated that age,tumor location,diameter of tumor, Borrmann type, type of histology, TNM stage, depth of infiltration, lymph node metastasis, liver metastasis, peritoneal dissemination, blood of transfusion during operation, extent of the radical cure of the tumor and excision techniques were significantly influential factors for the prognosis of patients. Cox regression analysis showed that tumor location, diameter of tumor,depth of infiltration, lymph node metastasis,liver metastasis, TNM stage, peritoneal dissemination, blood transfusion during operation, extend of the radical cure of the tumor and excision techniques were independent factors influencing the postoperative survival rate.Conclusion Independent factors influencing the postoperative survival rate include tumor location, diameter of tumor, lymph node metastasis, infiltration depth of the tumor, pathological classification, liver metastasis, peritoneal dissemination, and TNM stage, extent of the radical cure of the tumor, lymphanodectomy techniques and blood transfusion during operation are also important factors.

BACKGROUND:BIOSSN4 nickel-free stainless steel is an austenitic medical stainless steel material, which has passed the standard hemolysis test, cytotoxicity assays and sensitization test of the National Institute for the Control of Pharmaceutical and Biological Products. OBJECTIVE:To evaluate theinvitro cytotoxicity and corrosion resistance of a new medical BIOSSN4 nickel-free stainless steel. METHODS:The L929 mouse fibroblasts suspension was seeded in 96-wel plates at a concentration of 1×108 /L, and were divided into five groups. BIOSSN4 nickel-free stainless steel extract, 316L stainless steel extract, gold aloy extract, lead material extract (positive control) and RPMI1640 medium (negative control) were added respectively. After 1, 2 and 3 days of culture, cel morphology was observed. The absorbance value in each group was determined using MTT assay. The relative cel proliferation rate was calculated. Toxicity grading was evaluated. In the simulated oral environment, the eletric potential of corrosion, current density of corrosion and polarization resistance of BIOSSN4 no-nickel stainless steel, 316L stainless steel and gold aloy were determined. RESULTS AND CONCLUSION:Within 3 days of culture, in lead material extract group, cels shrunk; the number of cels significantly reduced; the relative growth rate was lower than that in the other four groups (P < 0.05). In the other four groups, the cel morphology was good, and the relative growth rate was over 75%. The toxicity of BIOSSN4 nickel-free stainless steel extract, 316L stainless steel extract and gold aloy extract was grade 1. The toxicity of lead material extract was grades 2-3. These results demonstrate that BIOSSN4 nickel-free stainless steel has good biocompatibility. The corrosion resistance of BIOSSN4 nickel-free stainless steel is higher than that of the 316L stainless steel but lower than that of the gold aloys.

This study was aimed to establish an identification method for Spatholobus suberectus and its adulterants by near-infrared spectroscopy (NIRS). Near-infrared diffuse reflection spectroscopy (NIRDRS) spectra of different S. suberectus and its adulterants were acquired by using OPUS INDENT analysis software. NIRDRS spectra clustering analysis model and identification model were established and verified. The results showed that S. suberectus from dif-ferent regions and its adulterants were identified successfully by clustering analysis model and identification model. It was concluded that Spatholobus suberectus and its adulterants can be identified rapidly and non-destructively by NIRS.

AIM:To evaluate the correlation between microRNA-1284 (miR-1284) and gastric cancer, and to investigate the underlying mechanism.METHODS: The expression of miR-1284 was examined by real-time PCR in 63 gastric cancer ( GC) tissue samples and 63 non-malignant adjacent tissue samples.The correlation between miR-1284 and the clinicopathological feature of GC was analyzed.Lentiviral vector containing miR-1284 was constructed and transfected into GC SGC-7901 cells.After transfection, the expression of miR-1284 was examined by real-time PCR.The cell activity was evaluated by CCK-8 assay.The cell cycle and apoptosis were determined by flow cytometry.The ability of cell migra-tion was measured by wound-healing assay.The potential target gene of miR-1284 was predicted by online bioinformatic softwares.The expression of JAG1 mRNA was examined by real-time PCR.The protein levels of JAG1, Notch1 and NF-κB were analyzed by Western blotting.RESULTS:Compared with non-malignant adjacent tissue samples, the results of real-time PCR showed significant downregulation of miR-1284 in 42 GC tissue samples ( P<0.05 ) .The expression level of miR-1284 was not significantly associated with age and gender of the patients, tumor size, TNM staging and lymph node metastases (P>0.05), but significantly associated with histologic grading (P<0.05).Compared with LV-NC-GFP group and control group, after transfection of miR-1284 in LV-miR-1284 group, the expression of miR-1284 was significantly in-creased (P<0.05), the percentages of apoptotic cells and the cells in G0/G1 phase were significantly increased (P<0.05), the cells activity and ability of migration were significantly decreased (P<0.05), and the expression of JAG1, Notch1 and NF-κB was significantly decreased (P<0.05).CONCLUSION:The inhibitory effect of miR-1284 on gastric cancer may be associated with the regulation of its targeting gene JAG1.

Objective To investigate the expression of microRNA-106b(miR-106b)in the placentas of patients with pre-eclampsia and its relationship with matrix metallopeptidase(MMP)-2,and its effect on the invasion and proliferation of trophoblasts. Methods (1) Placental tissues were collected from patients with mild pre-eclampsia (mPE, n=30) , severe pre-eclampsia (sPE, n=30) and normal pregnant women (n=40). Human choriocarcinoma cell lines JAR and JEG3 were assigned to the miR-106b mimics group, the mimics negative control group, the miR-106b inhibitor group and the inhibitor negative control group, respectively. (2) The target gene of miR-106b(such as MMP-2) was predicted by bioinformatics. Dual-luciferase reporting system was used to verify the regulation of miR-106b on the expression of MMP-2. (3) The expressions of miR-106b and MMP-2 were measured by quantitative real-time PCR (qRT-PCR) and western blot. (4) Cell proliferation was determined by MTT assay. (5) Invasive activities in each group were assessed by cell transwell invasion assays. Results (1) Predicting result of bioinformatics indicated that MMP-2 was one of the target genes of miR-106b. Dual-luciferase activity assay demonstrated that MMP-2 was the direct target of miR-106b(P＜0.01).(2) The results of qRT-PCR.①The expression of miR-106b in the placentas of mPE, sPE, normal pregnant women were 2.89±0.04, 1.96±0.03, 1.01±0.03, respectively (P＜0.05). And the expression of MMP-2 mRNA in the placentas of mPE, sPE, normal pregnant women were 1.87±0.05, 0.69±0.03, 2.78±0.03, respectively (P＜0.05).②The expression of miR-106b in the JAR cell line in the miR-106b mimics group, the mimics negative control group, the miR-106b inhibitor group and the inhibitor negative control group were 2.39 ± 0.03, 1.03 ± 0.04, 0.73 ± 0.03, 1.11 ± 0.04, respectively (P＜0.05). And its expression in the JEG3 cell line were 2.17±0.04, 1.18±0.04, 0.61±0.03 and 1.22±0.03, respectively (P＜0.05). ③The expression of MMP-2 mRNA in the JAR cell line in the miR-106b mimics group, the mimics negative control group, the miR-106b inhibitor group and the inhibitor negative control group were 0.45±0.15, 1.02±0.03, 2.28±0.03, 1.11±0.03, respectively (P＜0.05). And its expression in the JEG3 cell line were 0.58±0.03, 1.25±0.15, 2.25±0.03, 1.21±0.03, respectively (P＜0.05). (3) The results of western blot.①The expression of MMP-2 protein in the placentas of mPE, sPE, normal pregnant women were 1.63 ± 0.04, 0.55±0.03, 2.82±0.03, respectively (P＜0.05).②The expression of MMP-2 protein in the JAR cell line in the miR-106b mimics group, the mimics negative control group, the miR-106b inhibitor group and the inhibitor negative control group were 0.41 ± 0.03, 0.97 ± 0.03, 2.25 ± 0.03, 1.01 ± 0.03, respectively (P＜0.05). And its expression in the JEG3 cell line were 0.53±0.03, 1.20±0.03, 2.31±0.04, 1.19±0.03, respectively (P＜0.05). (4) miR-106b could inhibit the proliferation of JAR and JEG3 cells, cell proliferation rates in the miR-106b mimics group were lower than that in the mimics negative control group (P＜0.05). And cell proliferation rate in the miR-106b inhibitor group was higher than the inhibitor negative control group (P＜0.05) . (5) The numbers of JAR cell that passed the membrane in the miR-106b mimics group, the mimics negative control group. The miR-106b inhibitor group and the inhibitor negative control group were 61±15, 79±13, 134±13, 80±12, respectively( P＜0.05). And the numbers of JEG3 cell that passed were 57±12, 71±15, 128±15, 70± 14, respectively (P＜0.05). Conclusion The miR-106b could inhibit the invasion and proliferation of JAR and JEG3 cells through targeting MMP-2, and have a relationship with the pathogenesis of pre-eclampsia.

Objective To evaluate the quality of Radix et Caulis Ilicis Asprellae from Pingyuan planting base and Chinese herbal medicine market. Methods The water- and alcohol-soluble extracts from 19 batches of Radix et Caulis Ilicis Asprellae medicinal materials were detected according to Appendix ⅨH, ⅩA of the Chinese Pharmacopoeia ( 2010 edition). And the quality of the medicinal materials was evaluated by microscopic identification technology according to the method for Radix et Caulis Ilicis Asprellae recorded in Guangdong Provincial Chinese Medicine Standard, and then thin layer chromatography ( TLC) was optimized to establish the high performance liquid chromatography (HPLC) fingerprint. The HPLC was performed on Waters XBridgeTM C18 column (250 mm × 4.6 mm, 5μm) with acetonitrile(A)-0.2% (v/v) phosphorus acid (B) as the mobile phase by gradient elution, flow rate was 1.0 mL/min, and detection wavelength was 220 nm. Results The results of sample characters, TLC and microscopic identification showed that the samples of Radix et Caulis Ilicis Asprellae in Chinese herbal medicine markets were certified products, but stems and roots were blended. Seven common peaks were showed by HPLC and confirmed by similarity analytical software. The similarity of 15 batches of planting base samples was all above 0.9. Of 19 batches of the commercial samples, the similarity of 11 batches was above 0.9. The alcohol-soluble extract contents were in the range of 64.55 mg/g to 186.18 mg/g. Conclusion The medicinal materials of Radix et Caulis Ilicis Asprellae from Chinese herbal medicine market are certified products, but the qualities vary greatly for the blending of stems and roots and inadequate growth years. The quality of materials from planting base is better. The established method is helpful for the quality evaluation and control of Radix et Caulis Ilicis Asprellae.

Objective To establish the fingerprints and formononetin content determination method for Caulis Spatholobi from different habitats by high performance liquid chromatography ( HPLC) , thus to control the quality of Caulis Spatholobi. Methods Reversed phase-high performance liquid chromatography (RP-HPLC) for fingerprint was performed on Feini Gen RedClassical AQ-C18 column ( 4.6 mm × 250 mm, 5 μm) with acetonitrile-0.1%acetic acid solution as the mobile phase by gradient elution, and the detection wavelength was 260 nm. High performance liquid chromatography-diode array detector ( HPLC-DAD) for the determination of formononetin content was performed on AcclaimTM 120-C18 column ( 4.6 mm × 250 mm, 5 μm) with acetonitrile-water solution by isocratic elution, the detection wavelength was 254 nm, the flow rate was 1.0 mL/min and the column temperature was 25℃. Results The standard fingerprint of Caulis Spatholobi was set up through the evaluation of the fingerprints of 24 batches of Caulis Spatholobi samples from different habitats. Thirteen common peaks were identified with reference to formononetin peak, and the content of formononetin was determined by HPLC-DAD method. The similarity of the fingerprints of Caulis Spatholobi from different habitats and their formononetin content had great differences. Conclusion The established method is simple, accurate, highly sensitive, and repeatable, and can be applied for the quality control of Caulis Spatholobi.

Objective To investigate the effect of herpes simplex virus thymidine kinase/ganciclovir (HSV-TK/GGV)system driven by human alpha fetoprotein(AFP)enhancer on hepatocellular carcinoma(HCC)cells in vitro and in vivo.Methods HCC-specific eukarotypic expression vector carrying suicide gene driven by AFP enhancer(pAFP-cDNA3.1-TK)was constructed.The plasmid was trasfected to AFP-positive HepG2 cells and AFP-negative SMMC7721 cells by liposomes.Protein and mRNA expressions of TK were detected by RT-PCR or Western blot.The survival rates of HCC cells were detected by methyl thiazolyl tetrazolium assay.The effects of GGV on the in vitro proliferation,survival and apoptosis of HCC cells were observed,and the inhibitive effect of GGV on the survival of HCC cells in vivo was also detected.All data were analyzed by using the t test.Results The pAFP-cDNA3.1-TK was successfully constructed and transfected to the HCC cells.The protein and mRNA expressions of TK were detected in AFP-positive HepG2 cells.GGV dose-and time-dependently inhibited the growth and induced the apoptosis of HepG2 cells in vitro,but it had no effect on SMMC7721 cells.No protein or mRNA expression of TK was detected in the SMMC7721 cells.There was a significant difference on the inhibitory effects of GGV on HepG2 cells and SMMC7721 cells(t =2.58,2.73,3.12,P ＜0.05).GGV specifically inhibited the growth of AFP-positive HepG2 cells,and the inhibition rate was 46%;the growth of AFP-negative SMMC7721 cells was not influenced by GGV.There was a significant difference in the inhibitive effect of GGV on the growth of HepG2 cells and SMMC7721 cells(t = 3.36,P ＜ 0.05).Conclusion HSV-TK/GGV systemdriven by human APF enhancer kills APF-positive HCC cells and inhibits the growth of HCC cells.