The killing effects of herpes simplex virus thymidine kinase gene/ganciclovir (HSV-tk/GCV) approach by the addition of several commonly clinical chemotherapeutic agents on hormone refractory prostate cancer (HRPC) cells PC-3m were investigated. After transferring of the HSV-tk gene into PC-3m cells, mRNA and protein expression of HSV-tk was detected by reverse-transcript polymerase chain reaction (RT-PCR) and strept avidin-biotin complex (SABC) immunohistochemical method. The killing effect of GCV, cisplatin (CDDP), etoposide (VP-16), vincristine (VCR), methotrexate (MTX), 5-fluorouracil (5-Fu), and suramin on PC-3m cells was evaluated by morphological assessment analysis, trypan blue exclusion assay and MTT assay respectively. Additionally, the cooperative effect of HSV-tk/GCV system combined with the above agents on the target cancer cells was determined by MTT. Furthermore, apoptosis and necrosis induced by GCV plus 5-Fu or suramin was analyzed by flow cytometry (FCM). The results showed that that there was HSV-tk mRNA and protein expression in pDR2-tk plasmid transduced PC-3m cell. Combination of GCV with VP-16, VCR, 5-Fu or suramin led to an enhanced cellular killing effect, but with CDDP resulted in a reduced one and with MTX in an approximate one. FCM revealed that synergistic use of GCV and 5-fu or suramin resulted in a rather large proportion of apoptosis and necrosis with the apoptosis index being 36.38% and 35.51%, and the proportion of necrosis being 33.05% and 28.87%, respectively. In conclusion, HSV-tk/CGV approach by addition of certain clinical available chemotherapeutic drugs brings on statistically significant enhanced cell killing over single-agent treatment. Our results highlight the potential for such new combination therapies for future treatments of HRPC.

Connexin-43 (Cx43) expression in prostate cancer (PCa) cells and the potency of gap junctional intercellular communication (GJIC) in the cells were investigated, with an attempt to elucidate the reason why the so-called "bystander effect" mediated by thymidine kinase (TK) suicide gene therapy on PCa cells is not of significance and to explore the role of GJIC in PCa carcinogenesis. mRNA and protein expression of Cx43 in a PCa cell line PC-3m was detected by reverse-transcription polymerase chain reaction (RT-PCR) and strapt-avidin-biotin-enzyme complex (SABC) immunohistochemical staining, and inherent GJIC of PC-3m cells was assayed by scrape-loading and dye transfer (SLDT) assay. The expression of Cx43 in human normal and malignant prostate tissues was determined by SABC immunohistochemistry as well. It was found that Cx43 mRNA and protein expression in PC-3m cells was slightly reduced as compared with positive controls and the location of Cx43 protein was aberrant in cytoplasm rather than on membrane. Assessment of paraffin sections demonstrated that the expression of Cx43 protein in PCa cells was abnormally located and markedly diminished as compared with normal prostatic epithelial ones, displaying a negative correlation to the pathological grade (chi2=4.025, P<0.05). Additionally, capacity of inherent GJIC in PC-3m cells was disrupted, which was semi-quantified as (+) or (-). It was indicated that both down-regulated expression of Cx43 mRNA and aberrant location of Cx43 protein participated in the mechanisms leading to deficient GJIC in PC-3m cells. Lack of efficient GJIC is a molecular event, which may contribute not only to limited extent of "bystander effect", but also to initiation and progression of prostatic neoplasm.

AIM: To probe into the genetic susceptibility of HLA-DRB1 alleles to esophageal neoplasm in Hubei Han Chinese. METHODS: HLA-DRB1 gene polymorphism in 42 patients with esophageal neoplasm and 136 normal control subjects was studied by PCR and sequence. RESULTS: Allele frequency of HLA-DRB1 *0901 allele was significantly higher in esophageal cancer patients than those in normal controls (0 2500 vs 0 1397,P =0 028; the odds ratio 2 053; etiologic fraction 0 1282).There were no association between the rested HLA-DRB1 alleles with patients. CONCLUSION: Individuals carrying HLA-DRB1 *0901 may be susceptible to esophagealo carcinoma, its nucleotide sepuence approachs to the corresponded allele sequence(exon 2)published in GenBank.

Objective To explore the effect of Melatonin(MT) on CD4+CD25+ regulatory T cell (CD4+CD25+Tr)and airway inflammation in asthmatic rat.Methods Thirty-two SD rats were randomly divided into 4 groups,8 rats in each group.Asthmatic group:rats were immunized on day 1 and 7 by intraperitoneal inject of mixture of ovalbumin(OVA) and aluminumhydroxide.From day 14,the animals were allenged with aerosolized OVA for 20 min per day for 7 consecutive days.MT group:OVA-sensitized rats were injected intraperitoneally with 0.1 mg/kg MT 30 min before each OVA challenge.Dexamethasone group:OVA-sensitized rats were injected intraperitoneally with 0.5 mg/kg Dexamethasone 30 min before each OVA challenge.Control group:OVA for inhalation and MT for intraperitoneal injection was replaced with saline.After the last challenge,peripheral blood was stained to count the percentage of eosinophil(EOS).Then the rats were lavaged and total leukocytes counts in bronchoalveolar lavage fluid(BALF) were performed after staining with Wright-Giemsa staining.The EOS counts around the airway was counted after the histological section of lung staining with hematoxylin and eosin staining.The serum level of immunoglobulin E(IgE) was detected by immunoenhancement.The change of CD4+CD25+Tr was assessed with flow cytometry.SPSS 10.0 software was applied to analyze data. Results In asthmatic rats,the CD4+CD25+ Tr/ CD4+T cells ratio had significant negative relationship with the EOS counts around the airway and the total leukocytes counts in BALF (r=-0.73 P0.05).There was a significant decrease in the percentage of the eosinophils in peripheral blood,the eosinophil counts around the airway,the total leukocytes counts in BALF and the serum level of IgE in MT group compared with asthmatic group (Pa

OBJECTIVE: A systematic study of Laggera pterodonta was performed in order to identify its bioactive compounds. METHODS: Solvent partition and column chromatography used to isolate and purify Laggera pterodonta compounds. The isolated compounds were further elucidated by high field NMR spectroscopy. RESULTS: Fifty-two compounds were separated and the structures of 39 compounds were elucidated. Twenty-four compounds have not been previously reported. For example, the structure of compound 19, a previously undetected compound was elucidated as 4gamma, 9alpha,11-triol-enantio-eudesmane using both 1D and 2D NMR spectroscopy. Major secondary metabolites of the Laggera pterodonta included eudesmane-type sesquiterpenes and flavone derivatives. CONCLUSION: A systematic study of a yunnan herbal medicine Laggera pterodonta resvealed several novel compounds that may have clinical significance. Further in vivo animal studies should be performed to assess their bioactive role.

Objective To clarify the participation of oxidative stress in the white matter lesions induced by chronic cerebral hypoperfusion in the rats.Methods Chronic cerebral ischemia models were established by permanent occlusion of bilateral common carotid arteries(2-VO)in male Wistar rats.The rats were assigned to 5 groups(n=6):those with chronic cerebral ischemia for 3 and 7 d,3 and 6 weeks,and those given sham operation.We examined the activities of superoxide dismutase(SOD)and catalase(CAT),the giutathione(GSH)content and the changes of malondialdehyde(MDA)level and HNE modified protein in the white matter of rats.Results The MDA level of the hypoperfused rats was significantly increased 3 weeks after the operation as compared with that of the sham-operated rats with a further increase at 6 weeks(P<0.05).The HNE modified protein level in the hypoperfused rats was gradually and significantly increased from 3 d to 6 weeks after the operation as compared with that of the sham-operated rats(P<0.05).SOD activity of the hypoperfused rats was significantly decreased 3 and 6 weeks after the operation compared with that of the sham-operated rats(P<0.05),while CAT activeity was not altered.Moreover,the GSH content in the hypoperfused rats was gradually and significantly decreased 7 d and 6 weeks after the operation in comparison with that in the sham-operated rats(P<0.05).Conclusion Chronic cerebral ischemia results in increased oxidative damage in the white matter and decreased antioxidant defense capability,which is closely correlative to white matter lesions induced by chronic cerebral ischemia.

In this study, an analytical method was developed and used to quantify simultaneously protocatechuic acid, neochlorogenic acid, cryptochlorogenic acid and 1, 3-dicaffeoylquinic acid--four bioactive compounds contained in Fructus Xanthii using UPLC. The contents of four phenolic components of 28 batches of samples collected from different product areas and markets were determined and compared by means of this established method. The mobile phase was composed of methanol and water containing 0.1% phosphoric acid. Chromatography was monitored at dual-wavelengths--220 and 327 nm. Flow rate was 0.4 mL x min(-1) and column temperature was 35 degrees C. The correlation coefficient between concentration and chromatographic peak area of protocatechuic acid, neochlorogenic acid, cryptochlorogenic acid and 1, 3-dicaffeoylquinic acid was over 0.9999 in the range of 0.3570-35.70, 2.500-250.0, 1.060-106.1, 1.010-101.0 microg x mL(-1), respectively. The average recoveries of the four compounds were 97.68%, 99.55%, 97.92% and 100.4%, respectively. In conclusion, the established method can rapidly attain an accurate and reproducible result used to control the quality of Fructus Xanthii.
Chlorogenic Acid ; analysis ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Fruit ; chemistry ; Hydroxybenzoates ; analysis ; Plants, Medicinal ; chemistry ; Quality Control ; Quinic Acid ; analogs & derivatives ; analysis ; Reproducibility of Results ; Xanthium ; chemistry

This study was aimed to investigate the effect of multikinase inhibitor sorafenib on the proliferation and apoptosis of U937 cells and its possible mechanism. U937 cells were treated with different concentrations of sorafenib for 48 hours. Cell viability was determined by Cell Counting Kit-8; cell apoptosis and cell ratio in cell cycle were detected by flow cytometry with Annexin V/PI staining and PI staining respectively; expressions of GSK-3β, β-catenin and cyclin-D1 were assayed by Western blot. The results showed that the proliferation of U937 cells was inhibited by sorafenib in a dose-dependent manner (p < 0.05). Sorafenib induced cell apoptosis and cell cycle G(1)/G(0) arrest. Compared with results of Western blot before treatment, expression of inactivated GSK-3β, β-catenin and Cyclin-D1 down-regulated in a dose-dependent manner after treatment with sorafenib, this same changes were observed after up-regulation of inactivated GSK-3β by LiCl (p < 0.05). It is concluded that sorafenib inhibits the proliferation of U937 cells and induces cell apoptosis through reducing negative regulation of WNT signal pathway on inactivated GSK-3β and down-regulating β-catenin and cyclin-D1 level, which result in U937 cell cycle G(1)/G(0) arrest.
Apoptosis ; drug effects ; Benzenesulfonates ; pharmacology ; Cell Proliferation ; drug effects ; Cyclin D1 ; metabolism ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Humans ; Niacinamide ; analogs & derivatives ; Phenylurea Compounds ; Pyridines ; pharmacology ; U937 Cells ; Wnt Signaling Pathway ; beta Catenin ; metabolism

Objective:To compare clinical efficacy after endoscopic submucosal dissection(ESD)of gastric cancer recurrence with laparoscopic and open radical resection.Methods:ESD postoperative recurrence of 98 cases of gastric cancer were studied.According to different surgical methods were divided into observation group (49 cases of laparoscopic radical resection) and control group (49 cases of radical open surgery),3 years of follow-up,compared two groups of the curative effect,intraoperative and postoperative related indicators,postoperative,and the postoperative complications.Results:The two groups after 1～3 year survival rate and recurrence rate compared with no significant difference (P＞0.05).The operation time of the observation group was longer than the control group,but the total length and amount of bleeding were less than the control group,the differences were statistically significant (P＜0.05).The observation group of exhaust time,gastrointestinal decompression extubation time,extubation time of abdominal drainage,eating for the first time,hospitalization time and hospitalization expenses were less than the control group,the differences were statistically significant (P＜0.05).The patients in the observation group of the overall complication rate was 12.24％,and the control group (8.16％) there was no statistically significant difference (P＞0.05).The observation group patients with laparoscopic radical surgery and recurrence of gender,age,tumor staging,there were no obvious relationship (P＞0.05).The observation group of laparoscopic radical recurrence in patients with lymph node metastasis was 15 cases after surgery,but with gender,age,tumor staging,there were no obvious relationship (P＞0.05).Conclusions:The curative effect of ESD postoperative recurrence in patients with gastric cancer underwent laparoscopic surgery,less damage to patients,is conducive to the recovery,and high safety,worthy of promotion.

AIM: To report the long- term clinical outcomes of accelerated trans-epithelial corneal cross-linking ( CXL ) protocols using KXL System ( Avedro, USA ) in the treatment of progressive keratoconus. · METHODS: Totally 52 patients ( 102 eyes ) with progressive keratoconus between December 2014 and February 2017 [ maximum keratometry values ( Kmax) ≤60.0D, minimum corneal thickness(Thk) ≥400m] were treated with an accelerate trans-epithelial CXL protocol (UV-A irradiation intensity 45mW/cm2 with a total fluence of 7. 2J/cm2 ) using KXL system ( Avedro, USA ) in Southwest Hospital. The average follow-up time was 11. 65mo (range: 9-26mo). Uncorrected distance visual acuity ( UDVA) , corrected distance visual acuity ( CDVA) , intra- ocular pressure ( IOP ) , slit-lamp microscope examination, Kmax and average keratometry values ( AveK ) , corneal stromal demarcation line depth and endothelial cell density ( ECD) were evaluated. ·RESULTS:The 52 patients (102 eyes) were included in this research, male 36 (70 eyes) and female 16 (32 eyes), average age was 19. 5±4. 6 years. Preoperative CDVA was 0. 84±0. 89 (LogMAR), postoperative CDVA was 0. 69±0. 72 ( P = 0. 398 ). Preoperative UDVA was 1. 02 ± 0. 62 (LogMAR), postoperative UDVA was 0. 85 ± 0. 59 ( P =0. 154 ). Preoperative IOP was 12. 95 ± 4. 40mmHg, postoperative IOP was 11.92±3. 66mmHg (P=0. 272). No statistical difference (P=0. 552) has been found between preoperative and postoperative ECD. Nevertheless, on the Sirius anterior system ( Sirius, CSO, Itlay) , significant statistical difference (P=0. 017) was confirmed between preoperative Kmax ( 50. 83 ± 3. 48D ) and postoperative Kmax (52. 05±3. 63D). Meanwhile, the postoperative Avek (47.74±2. 51D) was significantly lower (P=0. 041) than the preoperative Avek ( 48. 73 ± 4. 33D ). The average corneal stromal demarcation line depth ( 192 ± 23. 6μm ) was detected by the anterior segment OCT. No statistical difference ( P = 0. 816 ) has been found between preoperative and postoperative Thk. No severe complication was observed in all cases. ·CONCLUSION: Accelerated trans-epithelial CXL was effective in decreasing keratometry values for progressive keratoconus in this research, and the outcomes remained stable during the follow-up time. No endothelium damage or other severe complications were observed in this clinical research. The accelerated trans-epithelial CXL is as effective as the standard CXL.