Objective To observe the preliminary effect and safety of perioperative hyperthermic intraperitonral chemotherapy to treat gastrointestinal cancer. Methods The clinical data of 60 patients with progressed gastric or colorectal carcinoma who underwent perioperatively HIPEC in our hospital from May, 2012 to October, 2013 were retrospectively analyzed. The incision healing, complications, KPS scores and serum CEA levels were observed. Results The vital signs of all patients were normal during HIPEC. There was no perioperative death. No patients underwent serious complications like diffuse peritonitis, intestinal obstruction, gastrointestinal perforation or intraperitoneal bleeding. There was no anastomotic leakage in 28 patients who underwent Stomach-jejunum anastomosis or intestinal anastomosis. After HIPEC, the life quality was improved;increase in KPS score and reduction in serum CEA levels were noted in all patients (P < 0.01). Of 29 patients with malignant ascites, 20 cases achieved complete mitigation and 8 cases achieved partial mitigation, 1 case was in stable condition, thus yielding effective rate of 96.5%. Conclusions It is safe and feasible for HIPEC to treat gastrointestinal cancer. HIPEC can improve the patients’ life qualities, without theincrease in perioperative complications. The short-term effect of HIPEC is confirmed in alleviating ascites.

Objective To explore an effective method of isolating and culturing the vascular endothelial cells of rat thoracic aorta.MethodThe thoracic aorta was harvested under aseptic condition from the thorax of a Wistar rat.The peripheral connective tissue and fat of the thoracic aorta were stripped and disposed,and then the thoracic aorta was turned inside out to expose the intima.The thoracic aorta was ligated with silk and cauterized on both ends with heated forceps.Then the thoracic aorta was cultured in medium DMEM/F12 containing 20% newborn calf serum in a 50 ml culture bottle which was already blanketed with rat tail collagen.The thoracic aorta was discarded and the new culture medium was added into the culture bottle six days later.The migrating cells were differentially digested by 0.125% pancreatic enzyme for serial subcultivation.The cells were identified by immunohistochemical method with anti-Ⅷ factor antibody.ResultA small amount of cells were seen to migrate from the aorta and adhered to the bottom of culture bottle 6 days after cultivation;the migrating cells spread to cover most part of the bottom of culture bottle 12-14 days later.About 70% of the migrated cells were in a confluent monolayer.The confluent cells grew rapidly after being digested with pancreatic enzyme,and they showed a typical cobblestone appearance.The cells were identified as endothelial cells with 100% expression of Ⅷ factor,which was regarded as the marker of endothelial cells.ConclusionThe method established in the present study is simple and easy to handle,it does not need collagen enzyme and endothelial cell growth promoting substrate,and it is economical and applicable.It is especially suitable for isolation and cultivation of vascular endothelial cells of vessels of small caliber.

OBJECTIVE:To establish a method for the decoction of isoflavone content and its acid hydrolysis conversion rate in the flower of Pueraria lobata. METHODS:Using the flowers of Pueraria lobata as raw material,the isoflavone with main com-ponent of tectoridin in the flower of P. lobata was prepared with ethanol,ethyl acetate extracted,ethanol recrystallized and puri-fied,and it was converted to tectorigenin with hydrolysis in hydrochloric acid. By screening the solvent and wavelength,UV spec-trophotometry was established to determine tectovidin and tectorigenin,and calculate the isoflavone content and acid hydrolysis con-version rate of tectoridin(expressed by the relative percentage of tectorigenin). It was compared with HPLC detection results,the accuracy of UV method was evaluated. RESULTS:The solvent was 70％ethanol solution containing 1％triethylamine,and the iso-flavone content was detected at wavelength of 339,274 nm. The linear range of tectoridin was 8.80-29.33 nmol/mL(r=0.9999). RSDs of precision(n=6),stability(n=5)and reproducibility(n=6)tests were lower than 1.94％;average recovery was 99.7％(RSD=1.77％,n=9). There were no statistical significances in the contents of total flavonoids (UV:17.64-25.55 nmol/mL vs. HPLC:17.39-24.40 nmol/mL) and the relative percentage of tectorigenin (UV:57.65％-87.59％ vs. HPLC:55.62％-91.14％). CONCLUSIONS:The established method is accurate,reliable,and can be used for the rapid determination of acid hydrolysis con-version rate of tectoridin.

Objective To investigate the modulation of ET-3 on the nuclear factor (NF)-κB/Bfl-1 antiapoptotic pathway in a malignant melanoma cell line A375. Methods Flow cytometry was performed to detect the apoptosis in cultured A375 cells after treatment with ET-3 of 100 nmol/L for 24 hours. ET-3 of various concentrations (0, 1, 10, 100 nmol/L) was used to treat some A375 cells with or without the pretreatment with the ETRB antagonist BQ788; after another 24-hour culture, RT-PCR and Western blot were conducted to examine the mRNA expression of Bfl-1 and protein expressions of Bfl-1 and ETRB, respectively. Results The 24-hour treatment with ET-3 of 100 nmol/L significantly reduced the apoptosis rate of A375 cells (F = 10.68, P ＜0.05). The mRNA and protein expressions of Bfl-1 were up-regulated by ET-3 in a concentration dependent manner (both P ＜ 0.01 ), while BQ788 significantly blocked the ET-3-induced up-regulation (F = 420.38,229.49, both P ＜ 0.01 ). The protein expression of pNF-κB in A375 cells was also enhanced by ET-3 of different concentrations (all P ＜ 0.05), but the enhancement was suppressed by BQ788, and there was a significant difference in the protein expression of pNF-κB between cells treated with ET-3 of 100 nmol/L and those treated with the combination of ET-3 of 100 nmol/L and BQ788 (F = 255.46, P ＜ 0.01 ). Conclusion ET-3/ETRB inhibits the apoptosis in A375 cells likely by activating the NF-κB/Bfl-1 anti-apoptotic pathway.

Objective:To establish an HPLC method for the determination of four kinds of isoflavone compounds including daid-zin, tectoridin, daidzein and tectorigenin in the extracts of Pueraria lobata flowers. Methods:The separation was performed on an Agi-lent C18 column(250 mm × 4. 6 mm, 5 μm) using a mobile phase of methanol-acetonitrile-water(2∶1∶2). The flow rate was 1. 0 ml· min-1 ,and the detection wavelength was 264nm. The column temperature was 25℃ and the sample size was 20 μl. Results:The de-tection could be accomplished within 10 minutes with good separation and specificity of four isoflavone compounds with the retention timeof3.4,3.8,5.7and7.2min,respectively. Thelinearrangewas0.784-78.440 μg·ml-1(daidzin),2.000-200.000 μg· ml-1(tectoridin) and 0.800-80.020 μg·ml-1 (daidzein and tectorigenin),and the relative coefficient was 0.999 9, 0.999 8, 0. 999 7 and 0. 999 9, respectively. The average recovery was 100. 54%(RSD=1. 66%,n=6),100. 03%(RSD=1. 00%, n=6), 99. 48%(RSD=1. 76%, n=6) and 100. 92%(RSD=2. 26%, n=6), respectively. Conclusion: The method is simple, rapid and accurate with good repeatability, which can be used in the rapid determination of isoflavone compounds in the flowers of Pueraria loba-ta.

Objective To optimize the extraction technology of flos Puerariae lobata isoflavone .Methods The flos Pu-erariae lobata isoflavone was distilled by ethanol circumfluence .Total flavonoids ,tectoridin and tectorigenin extracted from Puerariae using the UV and HPLC spectromertry methods were taken as evaluation indexes .Extraction technology was opti-mized with L9 (34 ) orthogonal test on the base of single observation of ethanol concentration ,solvent dosage and distilling time . Results The best extraction technology of flos Puerariae lobata isoflavone was :to add 12 times the amount of 70% ethanol for 90 minutes for the first time ,and 10 times the amount of 70% ethanol for 60 minutes for the second time .Conclusion The op-timized extraction process of flos Puerariae lobata isoflavone is reasonable and feasible ,and it can offer reference to actual pro-duction .

OBJECTIVETo observe the protective effect of tectorigenin on the vascular endothelial cells(VEC) injured by oxidant low density lipoprotein-cholesterol and the expression of MCP-1 and ICAM-1 mRNA, and explore the mechanism of anti-atherosclerosis.

METHODThe VEC of rat was cultured in vitro and the 100 mg x L(-1) ox-LDL inducing oxidant injured model was used in this study. Different dosage tectorigenin was added into VEC and the activity of VEC was observed by MTT colorimetry. The expression of MCP-1 and ICAM-1 mRNA in VEC was detected by RT-PCR.

RESULTTectorigenin had significantly protective effect on the VEC injured by ox-LDL and obviously inhibited the excessive expression of MCP-1 and ICAM-1 mRNA in VEC.

CONCLUSIONIt was the critical mechanism of anti-atherosclerosis that tectorigenin prevented the VEC oxidant injured and inhibited the excessive expression of MCP-1 and ICAM-1.

Animals ; Atherosclerosis ; drug therapy ; genetics ; metabolism ; Cells, Cultured ; Chemokine CCL2 ; genetics ; metabolism ; Disease Models, Animal ; Endothelial Cells ; drug effects ; metabolism ; Gene Expression ; drug effects ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; metabolism ; Isoflavones ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar

Objective:To explore the effect of teachers-standardized patients combined with case based learning (CBL) in clinical probation of gastrointestinal surgery.Methods:A total of 140 clinical medicine undergraduates in gastrointestinal surgery were selected as the research subjects, and were randomized into the intervention group and the control group, with 70 cases in each group. Traditional teaching method is performed in the control group, and teachers-standardized patients combined with CBL is conducted in the intervention group. Student knowledge assessment and satisfaction survey were made for teaching effect evaluation. The t test and chi-square test were performed by SPSS 24.0 for comparison between the two groups. Results:The medical records writing of the students in the intervention group and the results of the theoretical assessment [(83.20±7.94) and (82.74±7.19) points] were significantly higher than those of the control group [(79.57±9.26) and (79.49±7.86) points] ( P<0.05). Students in the intervention group have a higher level of satisfaction of this teaching mode. Conclusion:Teachers-standardized patients combined with CBL can improve teaching quality and learning enthusiasm in gastrointestinal surgery, and it is worthy of promotion and application in teaching.

【Objective】 To describe the distribution characteristics of knee and ankle joint range of motion and muscle atrophy related indexes in patients with Kashin-Beck disease (KBD) in Shaanxi Province so as to explore the correlation of knee and ankle joint range of motion (ROM) with muscle atrophy indexes and provide reference for clinical characteristics analysis of KBD patients. 【Methods】 To investigate the registered KBD patients from KBD areas in Shaanxi Province, we measured the general demographic data of the patients were collected and the ROM of the knee joint (flexion and extension), the ankle joint (dorsiflexion and plantar flexion), and the muscle atrophy related indexes such as the upper arm circumference, thigh circumference, calf circumference and grip strength. According to the population characteristics, i.e., gender, age, body mass index (BMI) and KBD grade, the median and quartile of joint ROM and muscle atrophy of KBD patients were reported, and then the differences in each index among different groups were analyzed. Partial correlation analysis was used to explore the correlation between indicators after controlling for variables such as gender, age and BMI. 【Results】 A total of 480 patients with KBD were investigated in this study, who consisted of 249 (51.9%) males and 231 (48.1%) females, with an average age of (63.10±7.32) years and an average BMI of (23.49±8.90) kg/m². The knee flexion ROM, knee extension ROM, ankle dorsiflexion ROM and ankle plantar flexion ROM were [105.0 (95.0, 120.0)] °, [0.0 (-15.0, 0.0)] °, [5.0 (0.0, 15.0)] ° and [20.0 (15.0, 30.0)] °, respectively, in KBD patients in Shaanxi Province. The left thigh circumference, right thigh circumference, left calf circumference, right calf circumference, and upper arm circumference were [43.0 (40.0, 47.0)] cm, [43.0 (39.0, 47.0)] cm, [29.0 (27.0, 32.0)] cm, [29.5 (27.0, 32.0)] cm, [27.0 (25.0, 30.0)] cm, respectively. The left hand grip strength and right hand grip strength were [13.4 (9.5, 18.4)] kg and [13.9 (9.8, 18.2)] kg, respectively. With the increase of age, the extension range of the left and right knee joints of KBD patients showed a decreasing trend (H=31.499, 31.847; all P<0.001). The range of motion of bilateral knee flexion was higher in the normal BMI group than in the overweight or obese group, with statistically significant differences (H=7.753, 12.333; P=0.021, 0.002). The knee flexion, thigh circumference, and calf circumference of the left and right sides showed a decreasing trend under different KBD grades (H=14.345, 17.256, 8.000, 8.462, 8.558, 9.633; all P<0.05). Correlation analysis showed that knee flexion ROM was positively correlated with thigh circumference, calf circumference, and grip strength in patients with KBD (all P<0.05). There was a positive correlation between knee extension ROM and thigh circumference in patients with KBD (P<0.01). 【Conclusion】 The impaired joint ROM and muscle atrophy are serious in KBD patients in Shaanxi Province, and there is a correlation between joint motion and muscle atrophy.