Objective To investigate the effects of combination of retinoids and interferon ?-2a on growth inhibition and apoptosis induction in bladder cancer cell lines. Methods Four bladder cancer cell lines and two retinoids,all-trans-retinoic acid (ATRA),9-cis retinoic acid (9cRA),combined with interferon ?-2a(IFN),were used in the study.The abilities of these agents to inhibit growth,induce apoptosis were compared and the expression of nuclear retinoid receptors,Stat1 protein were measured by means of TUNEL,FCM,RT-PCR and Western blot. Results Most of the bladder cancer cell lines were resistant to the effect of ATRA and 9cRA on growth inhibition and apoptosis induction.The effects of ATRA and 9cRA on cell growth and apoptosis were enhanced by IFN ?-2a.Combination of ATRA and IFN ?-2a induced RAR? and Stat 1 expression in three bladder cancer cell lines. Conclusions INF ?-2a synergizes with the inhibitory effect of ATRA and 9cRA on the growth inhibition and apoptosis of bladder cancer cells in vitro,up-regutation of Stat1 gene expression could be the main molecular mechanism.

Objective To investigate the feasibility and effectiveness of laparoscopic extraperitoneal inguinal hernia repair with low-pressure pneumoperitoneum without using stapling devices.Methods A total of 38 patients with reducible inguinal hernia underwent laparoscopic extraperitoneal inguinal hernia repair in our hospital from January 2006 to February 2007.Three abdominal trocars were introduced into the extraperitoneal cavity.An 11-mm trocar was placed at the lower border of the umbilicus,and the other two sized 5.5 mm were at the upper and lower 1/3 of the line between the umbilicus and the pubic symphysis,respectively.Then,low-pressure(6-8 mm Hg) CO2 was insufflated into the extraperitoneal cavity.Non-traumatic forceps was used to create an operative space from the hilum to the pubic symphysis and the diseased inguinal area.The hernia sac and the fabrics around it were disconnected.For big hernia sacs,the sac was ligated,cut at the cervix,and then left at its original site.While for small hernia sacs,it was dissociated and put back into the abdominal cavity.Afterwards,a patch sized 12 cm ? 15 cm was placed into the cavity,stretched,and attached to the abdominal wall,covering the annulus inguinalis profundus,Hesselbach triangle,and the femoral ring,without using stapling devices.After the operation,the patients received sandbag compression at the operative area and were kept in bed with urethral catheter for 24 hours. Results The operation was successfully accomplished in all the 38 cases without conversion to open surgery.The operation time was 45-85 min(mean,56 min).The intraoperative blood loss was 5-10 ml.After the operation,seroma occurred in 4 cases,and perforation in 2.No subcutaneous emphysema,hypercapnia,intestinal injury,neuralgia,or hemorrhage was found.The patients were followed up for 6 to 19 months(mean,10.5 months),and no recurrence occurred.Conclusions It is feasible to use laparoscopic extraperitoneal inguinal hernia repair with low-pressure pneumoperitoneum without using stapling devices.The method is associated with few complications and confirmed efficacy.

Objective To study the anti-inflammation and analgesic action s of Guanjiekang Tablet and to observe its toxicity.Methods Rat pedal swelling experiment,rat capillary permeability experiment,mouse acetic-acid-induced twisting experiment and mouse acute toxicity experiment were carried out in this study.Results Guanjiekang Tablet could obviously reduce the twisting times in mice(P 0.05).The LD50 of oral dose in mice is 1.74(g/kg).Conclusi ons Guanjiekang Tablet has good anti-inflammation and analgesic actions,but also has some toxicity.

Objective To study the feasibility,superiority and curative effect of percutaneous nephrolithotomy with fibrous sheath and Swiss LithoClast Master (EMS Ⅲ) through T-tube tract for retained choledocholithiasis. Methods This study involved 32 patients with hepatolithiasis who were treated in our hospital from August 2004 to August 2007. By combining percutaneous nephrolithotomy (with a fibrous sheath on) and Swiss LithoClast Master (EMS Ⅲ) through a T-tube tract,retained choledocholithiasis was removed. Results The 32 patients totally underwent 36 operations. The mean operation time was 43 minutes (ranged from 33 to 78 minutes). In 28 of the cases,the stones were extracted completely by one session; and 2 were cured by two operations. The final stone-free rate was 94% (30/32). Follow-up was carried out in all the patients for 26 to 48 months (mean,29 months),during which no patients developed abdominal pain,jaundice or fever,no recurrence was detected by B-ultrasonography. Two patients showed residual stones after the treatment,one of them received Roux-en-Y hepatocholangioenterostomy because of extensive multiple biliary stones in the liver,which could no be removed completely after three sessions of nephrolithotomy; another patient who was a 70-year old man refused the secondary operation and thus retained a few stones in the right inferior lobe of the liver. In this series of 32 cases,no bile duct tear,massive hemorrhage,biliary leakage,cholangitis or abdominal distension occurred. Conclusions It is a safe,convenient,and simple method to combine percutaneous nephrolithotomy with Swiss LithoClast Master via T-Tube tract for retained choledocholithiasis. The procedure results in less pain in patients,and reduced equipment spoilage,while the cost is low. As the surgery is easy to perform,it is worth being widely used.

[Objective] To optimize the technology for Antipyretic Mixture (AM), which is mainly composed of Herba Ephedrae, Gypsum Fibrosum (GF), Rhizoma Amenarrhenae, Radix Isatidis, Rhizoma Imperatae, Radix Puerariae, etc. . [Methods] Orthogonal test was applied for the screening of technology. Rat model of fever was induced by dried yeast and body temperature served as the screening index. [Results & Conclusion] The optimal process was as follow: (1) cleaning the herbs and smashing Gypsum Fibrosum into pieces and then soaking for one hour; (2) decocting the herbs with 10-fold water for one hour (Gypsum Fibrosum decocted first for one hour) ; (3) extracting the volatile oil, filtering and then concentrating the decoction; (4) evaporating again the volatile oil; (5) mixing the solutions and preparing as 1 kg/L oral liquid.

Objective To investigate the expression of HSPA9 in hepatocellular carcinoma(HCC) and its relationship with clinicopathological features and prognosis.Methods Forty-nine cases HCC treated by operative resection and follow up data in our hospital from January 2006 to January 2010 were retrospectively analyzed.Immunohistochemistry was performed to determine the expression of HSPA9 in HCC and paratumor tissues.The relationship between HSPA9 expression and clinicopathological features and prognosis was statistically analyzed.Results The HSPA9 protein expression in tumor tissue was higher that that in the paratumor tissue(t=6.601,P＜0.01),moreover the over expression of HSPA9 was significantly correlated with lymph node metastasis (P =0.005),TNM-stage(P =0.015),tumor differentiation (P =0.033),microvascular invasion (P =0.009) and recurrence (P =0.047).In the survival analysis results,the patients with over expression of HSPA9 had a much lower total survival rate(P=0.002)and much higher postoperative cumulative recurrence rate(P =0.003).There were significant differences in TNM-stage,microvascular invasion,lymph node metastasis,tumor differentiation and HSPA9 staining for overall survival and cumulative recurrence rate based on a univariate analysis(P＜0.05).Conclusion HSPA9 has over expression in HCC.The over expression of HS-PA9 is closely related to invasion and metastasis pathological features and can serve as an independent prognostic risk factor for predicting the prognosis of HCC.

OBJECTIVE: To study anti-inflammatory effect and mechanism of the luteolin · 4, 4' -dipyridy co-crystal.METHODS: Using macrophage RAW264. 7 of normal mice as control, the inflammation model was established with lipopolysaccharide (LPS) -induced RAW264. 7 cells. MTT assay was used to detect cells activity 2 h after treatment of different concentrations of luteolin (10, 20, 40, 80 μmol/L), 4, 4' -dipyridy (10,20, 40,80 μmol/L) and luteolin·4, 4' -dipyridy co-crystal (10, 20, 40, 80 μmol/L). The mRNA expression of iNOS and COX-2 in RAW264. 7 cells at 40 μmol/L were determined by qRT-PCR. The protein expression of TNF-α and IL-6 in RAW264. 7 cells at 40 μmol/L were determined by ELISA. The protein expression of NF-κB p65 in RAW264. 7 cells at 40 μmol/L were determined by Western bolt. RESULTS: Compared with normal cells, the activity of RAW264. 7 cells was decreased significantly after induced by LPS (P<0. 01); mRNA expression of iNOS and COX-2, protein expression of TNF-α, IL-6 and NF-κB p65 were increased significantly (P<0. 01). Both luteolin and luteolin · 4, 4' -dipyridy co-crystal could enhance the activity of RAW264. 7 cells after induced by LPS (P<0. 05 or P<0. 01) in concentration-dependent manner. 4, 4' -dipyridy had no significant effect on the activity of RAW264. 7 cells after induced by LPS. After luteolin and luteolin· 4, 4' -dipyridy co-crystal at 40 μmol/L, mRNA expression of iNOS and COX-2, protein expression of TNF-α, IL-6 and NF-κB p65 in RAW264. 7 cells after induced by LPS were decreased significantly (P<0. 05 or P<0. 01); the luteolin · 4, 4' -dipyridy co-crystal was better than luteolin (P<0. 05 or P<0. 01). CONCLUSIONS: The luteolin·4, 4' -dipyridy co-crystal can inhibit the generation of inflammatory factors by down-regulating NF-κB signal, and its anti-i nflammatory effect is better than luteolin.

Using the polymmerse chain reaction (PCR), we amplified the phytase gene phyA from Pichia pastoris GS115-phyA in Aspergillus niger NRRL3135 without the signal peptide sequence and intron sequence,. Then, it was cloned into pINA1297 vector to generate a recombinant vector of pINA1297-phyA. pINA1297-phyA was linearized and transformed into Yarrowia lipolytica po1h by the lithium acetate method. The positive transformants were obtained by YNB(casa) and PPB plates, after induced in YM medium at 28 degrees C for 6 day. The activity of the expressed phytase phyA reached 636.23 U/mL. The molecular weight of the enzyme was 130 kDa measured with SDS-PAGE analysis, whereas its molecular size reduced to 51 kDa after deglycosylation which is correspond with theoretical value. The enzymatic analysis of the recombinant phytase phyA revealed its optimal pH and temperature was 5.5 and 55 degrees C, which had high activity after incubated in pH ranged from 2.0 to 8.0 for 1 h. Moreover, its activity remained 86.08% after exposure to 90 degrees C for 10 min. It also was resistant to pepsin or trypsin treatment.
6-Phytase ; biosynthesis ; genetics ; Aspergillus niger ; genetics ; metabolism ; Pichia ; enzymology ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Yarrowia ; genetics ; metabolism

Microbial lipopeptides play an important role in apoptosis induction of tumor cells. However, there is little knowledge about the relationship between apoptosis induction and membrane fatty acids. The present study focused on the effects of lipopeptides produced by Bacillus subtilis HSO121 on Bcap-37 cell lines. 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl (MTT) colorimetric assay and surface tension measurements, showed that the critical micelle concentration (CMC) was a critical level for the inhibitory activity of lipopeptides on the growth of Bcap-37 cells. Under the CMC, the order of least to greatest cytotoxicity effect on cancer cell lines by lipopeptides is C(13)-lipopeptide < C(14)-lipopepitde < C(15)-lipopeptide. Above CMC, all lipopeptides directly exert cytolytic activity. The flow cytometric analysis and Hoechst33258 staining experiments confirmed the apoptosis of Bcap-37 cell lines induced by lipopeptides in a dose-dependent manner. This apoptosis was associated with a significant decrease of the unsaturated degree of the cellular fatty acids of Bcap-37 cell lines due to the changes in the cellular fatty acids composition induced by the lipopeptide treatment. These results indicated that disturbance of the cellular fatty acid composition of breast cancer cell lines were related to in the cell apoptosis. Furthermore, significant difference in IC(50) values of tumor cells and normal cell showed that the lipopeptide exerted selective cytotoxicity on the cancer cells. Thus HSO121 lipopeptides may have potential applications as an anticancer leads.
Antineoplastic Agents ; chemistry ; pharmacology ; Apoptosis ; Bacillus subtilis ; Cell Cycle ; Cell Membrane ; chemistry ; metabolism ; Cell Survival ; drug effects ; Fatty Acids ; chemistry ; metabolism ; HeLa Cells ; Humans ; Lipopeptides ; chemistry ; pharmacology ; Ploidies ; Tumor Cells, Cultured

Objective: To determine the HIV infection of a 13-year-old leukemia patient Wei using molecular tracing technique. Methods: Three blood samples were collected from the persons who were associated with HIV positive blood donation member Lang. The viral load was tested and pol gene was sequenced and analyzed. At the same time using HyPhy2.2.4 and Cytoscape 3.6.1 to establish the molecular network with these 3 samples sequences and other HIV subtypes sequences obtained from Luzhou. Results: The HIV-1 viral load test result of the three patients were: Lang 933 CPS /ml (treated, blood donor), Wei (blood recipitent) 89 813 CPS /ml, Deng (blood recipitent) 85 158 CPS /ml. The subtype of HIV-1 of these three samples were all recombinant HIV CRF01_AE, and the pol gene similarity was 98.8% (Lang and Wei), 99.7% (Lang and Deng), and 99.1% (Wei and Deng), respectively. The gene sequences of the three samples were linked into clusters (the gene distance was less than 0.004). Conclusions The patient Wei with leukemia was infected with HIV-1 due to blood transfusion during the seroconversion period, and the three patients were more likely to be infected with the same HIV-1 strain. It is necessary to popularize the use of high-sensitivity nucleic acid detection method in blood collection and supply institutions, which can effectively improve the safety of blood use.