Objective:To investigate the mechanism and effects of autophagy on cisplatin(DDP)-induced apoptosis in human gas-tric cancer cell line SGC7901. Methods:Cell proliferation was determined by an MTT assay after the SGC7901 cells were treated with DDP and/or chloroquine. Cell apoptosis was determined by flow cytometry. Autophagy and related protein expressions were detected by Western blot. Autophagy was quantitatively analyzed by fluorescence microscopy after monodansylcadaverine staining was per-formed. Results:The cells were treated with 5 mg/L of DDP for 24 h, the rate of cell apoptosis was (21.07 ± 2.12)%. Autophagy, char-acterized by an increase in the number of autophagic vesicles and LC3-II protein level, was observed in DDP-treated cells. After autoph-agy was inhibited by chloroquine, the rate of cell apoptosis was increased to (30.16 ± 3.54)%. In addition, caspase-3 and P53 protein levels were increased, but Bcl-2 protein was decreased. Conclusion:Autophagy protected human gastric cancer cell line SGC7901 from DDP-induced apoptosis. In addition, the inhibition of autophagy could promote apoptosis. The combined therapy of DDP and chlo-roquine may be a promising therapeutic strategy for gastric cancer.

Objective To evaluate the safety and effect of one-way barbs line (absorbable sutures v - locTM180) in primary suturing of laparoscopic choledocholithotomy. Methods From July 2014 to June 2015, clinical data of 86 cases performed primary suturing in laparoscopic choledocholithotomy by the same performer were retrospectively analyzed. The patients were divided into three groups, One-way barbs line continuous full-thickness suture group (A group), ordinary absorption line continuous full-thickness suture group (B group) and ordinary absorption line discontinuous full-thickness suture group (C group). The bile duct suture needed time, intraoperative blood loss, postoperative eating time, postoperative hospital stay and postoperative bile fistula were compared. Results All the 86 cases underwent successful operation without T tube, and none of them received alternative open operation. Compared with group B and C, time needed for bile duct suturing in A group have statistical significance. There was no significant difference in the intraoperative blood loss, postoperative eating time, postoperative hospital stay between the three groups (P > 0.05). The incidence of postoperative bile fistula in A group were none. The incidence of postoperative bile fistula in B group was 1 case, the incidence of postoperative bile fistula in C group was 2 cases.Conclusions The method of one-way barbs line continuous full-thickness suture were simple and secure in primary suturing of laparoscopic choledocholithotomy.

OBJECTIVETo evaluate the safety and effectiveness of GreenLight 120-W laser photoselective vaporization of the prostate (PVP) versus transurethral resection of the prostate (TURP) for benign prostatic hyperplasia (BPH).

METHODSWe searched PubMed, Medline, Embase, Cochrane Library, Wanfang, CNKI, and VIP for randomized control trials and their references addressing 120-W PVP versus TURP in the treatment of BPH. Based on the inclusion and exclusion criteria, two reviewers independently accomplished the screening, quality assessment, and data extraction of the identified studies and performed meta-analyses using RevMan 5.2.

RESULTSTotally, 6 randomized control trials were included in this analysis, involving 703 cases, 351 treated by PVP and 352 by TURP. Compared with TURP, PVP showed significantly decreased time of catheterization (by 32. 55 hours, 95% CI 15.3 -49.8, P < 0.01), hospital stay (by 1.85 days, 95% CI 1.2-2.5, P < 0.01), and intraoperative blood loss (by 15.6 g/L, 95% CI 10.0-21.2, P < 0.01), but increased time of operation (by 9.37 minutes, 95% CI 5. 1-13.6, P < 0.01). There was also a significant reduction in blood transfusion, TUR syndrome, and capsular perforation in the PVP group. At 12 months after surgery, no statistically significant differences were found between the two groups in the improvement of maximum urinary flow rate, IPSS, postvoid residual, and sexual function.

CONCLUSIONGreenLight 120-W laser PVP is a safe and effective procedure for the treatment of BPH, with similar effectiveness to TURP but less blood loss, shorter time of catheterization and hospital stay, and lower incidences of blood transfusion, TUR syndrome and capsular perforation.

Blood Loss, Surgical ; Humans ; Laser Therapy ; adverse effects ; methods ; Length of Stay ; Male ; Prostate ; surgery ; Prostatic Hyperplasia ; surgery ; Randomized Controlled Trials as Topic ; Treatment Outcome

Objective To explore the clinical application of percutaneous endoscopic gastrostomy in gastroin- testinal nutrition among critically ill patients.Methods Twenty-two patients were undertaken percutaneous endo- scopic gastrostomy under the lead of gastroscope.The puncturatio site was located in anterior wall of stomach.Results All the procedures were performed successfully for one time.Intraoperational blood pressure was very steady.At the same time,intraoperational SpO_2 of the patients all exceeced 97%.The bleeding amount and operation time were respectively (3.8?1.9) ml and (15.5?2.3) min.Severe complications such as gastrostoma,gastrocolic fistula, pneumoperitoneum,refluxing or aspiration of gastric juice,inhalant pneumonia didn't occur after the operation.Con- clusion Percutaneous endoscopic gastrostomy was safe and feasible among critically ill patients who needed gastroin- testinal nutrition because of its slight injury,little bleeding and shorter operational time.

BACKGROUNDRNA interference (RNAi) is a powerful tool to silence gene expression post-transcriptionally. Our previous study has demonstrated that small interfering RNAs (siRNAs) have sufficiently inhibited hepatitis B virus (HBV) replication and expression in vitro. In this study we observed the RNAi-mediated inhibitory effects on HBV replication in mice models and accessed the specificity of these effects.

METHODSA mutant RNAi vector (pSI-C mut) with two base pairs different from the original target gene sequence at the RNAi vector (pSI-C) was constructed according to the method described in this study. A mouse model of acute hepatitis B virus infection was established by injecting naked plasmid pHBV1.3 via the tail vein with acute circulatory overload. pSI-C, pSI-C mut and the irrelevant RNAi control plasmid for green fluorescent protein (GFP) gene, pSIGFP were respectively delivered with pHBV1.3 by tail vein injection method. Six days post injection, enzyme-linked immunosorbent assay (ELISA) assay was used to measure the concentration of HBV surface antigen (HBsAg) in mouse serum, immunohistochemical straining method was used to visualize the expression of HBV core protein (HBcAg) in liver tissues, and the transcriptional level of HBV C mRNA in liver tissues was detected by reverse transcriptase PCR (RT-PCR) analysis.

RESULTSInjection of pSI-C exerted magnificent and specific inhibitory effects on the replication and expression of HBV in the murine model. After 6-day post-injection (p.i.), the OD values were shown to be 5.07 +/- 1.07 in infecting group and 0.62 +/- 0.59 in pSI-C group. The concentration of HBsAg in pSI-C group was significantly lower than that in infecting group (P < 0.01). Liver intracellular synthesis of viral core protein was sharply reduced to 0.9% +/- 0.1%, compared with 5.4% +/- 1.2% of positive hepatocytes in infecting group (P < 0.01), and the transcriptional level of HBV C mRNA was greatly reduced by 84.7%. However, the irrelevant RNAi control plasmid (pSIGFP), and the pSI-C mut did not show the same robust inhibitory effects as pSI-C.

CONCLUSIONpSI-C exert efficient and specific inhibitory effects on HBV replication and expression in mice models.

Animals ; Hepatitis B ; therapy ; virology ; Hepatitis B Core Antigens ; biosynthesis ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; genetics ; physiology ; Male ; Mice ; Mice, Inbred BALB C ; RNA Interference ; RNA, Messenger ; analysis ; RNA, Small Interfering ; therapeutic use ; RNA, Viral ; analysis ; Virus Replication

Antiviral Agents ; pharmacology ; Dose-Response Relationship, Drug ; Hep G2 Cells ; Hepatitis B Surface Antigens ; metabolism ; Hepatitis B e Antigens ; metabolism ; Hepatitis B virus ; drug effects ; Humans ; Interferon-alpha ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Recombinant Proteins ; Reverse Transcriptase Polymerase Chain Reaction ; STAT1 Transcription Factor ; genetics ; metabolism

In this study the effects of red orpiment on NB4 and HL-60 cells were tried to determine. Semi-quantitative RT-PCR to determine the PML mRNA expression, immuno-fluorcscence study, together with the fluorescence stain and morphological observations were used. The results showed that: (1) red orpiment induces apoptosis morphologically in NB4 and HL-60 cells, the morphology of typical apoptosis can be seen in NB4 and HL-60 cells after 12 hours of treatment with red orpiment. Through the Wright's stain, we can see the extensive membrane blebbing, condensation of heterochromatin, and cell fragmentation, apoptotic body appearing. Many dead cells can be found on the second day. (2) in NB4 cells, red orpiment is shown to induce the PML-RARalpha chimera disappearance and to reorganize then to degradation of PML nuclear bodies which also seen in HL-60 cells, indirect immunofluorescence staining of PML with a specific monoclonal antibody was performed in control and treated cells. In NB4 cells, the control was diffusely microspeckled pattern of immunoreactivity. Upon red orpiment treatment, the microspeckled pattern disappeared, PML protein reversed into normal location. and the the size and the brightness of the particles were increased obviously. The normal nuclear distribution of PML protein was seen in untreated HL-60 cells. After treatment with red orpiment, in the nuclei of HL-60 cells, the size and the brightness of the particles were also increased. After two days of treatment with red orpiment, the immunofluorescent particles in cells almost disappeared. (3) the expression of PML mRNA is not changed in red orpiment-treated cells, RT-PCR to determine the PML mRNA expression in NB4 and HL-60 cells treated with red orpiment, the expression results are similar to the controls, that to say, the PML mRNA lever is unaffected. It was concluded that, red orpiment induced PML to play the effects of induce apoptosis in leukemia cells at the translational level and inhibited the proliferation of leukemia cells.

OBJECTIVETo investigate the anti-HBV effect of fusion protein thymosin alpha1-interferon alpha (TA1-IFN) in vitro and to compare its effect with a combination of interferon alpha and thymosin alpha1.

METHODSAfter 2.2.15 cells were seeded for 24 hours, drugs of five serial concentrations (8000, 4000, 2000, 1000, 500 U/ml) were added to the wells, then the medium was changed every three days. After 2.2.15 cells were treated with drugs for 6 days, the medium was collected. The inhibitory rates on HBsAg and HBeAg were determined using Abbot kit, and the cytotoxicity of different drugs by means of MTT colorimetric assays was also observed.

RESULTSThe inhibitory rate of fusion protein on HBsAg, HBeAg was dose-dependent and reached the maximum at 8000 U/ml concentration. In the meantime, the inhibitory rates of fusion protein on HBsAg and HBeAg were 72.2% +/- 0.8% and 60.4% +/- 1.1% respectively, and the cell survival rate was 85.2% +/- 2.0%; In the corresponding concentration, the inhibitory rates of combination thymosin alpha 1 and interferon alpha on HBsAg and HBeAg were 40.0% +/- 0.7%, 34.5% +/- 3.2% respectively. The results showed significant statistical differences between them; cell survival rate 70.0% +/- 1.9%, and the difference of the results was also significant. Cytotoxicity of fusion protein was weaker than a combination of thymosin alpha 1 and interferon alpha.

CONCLUSIONFusion protein TA1-IFN exerted stronger anti-HBV effects in vitro. Its anti-HBV effects in vitro were stronger than the combination of thymosin alpha and interferon alpha, and its cytotoxicity was weaker than the combination of thymosin alpha and interferon alpha. Our studies provided important evidence for clinical research on TA1-IFN, and also brought new hope for hepatitis B therapy.

Antiviral Agents ; pharmacology ; Hepatitis B virus ; drug effects ; Humans ; Interferon-alpha ; biosynthesis ; genetics ; pharmacology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; Thymosin ; biosynthesis ; genetics ; pharmacology

BACKGROUND AND OBJECTIVEThe primary submucous type of nasopharyngeal carcinoma (NPC) or the recurrent NPC in the parapharyngeal space is difficult to be diagnosed histologically by conventional biopsy because of the obstruction of the surrounding structures. This study was performed to evaluate the needle biopsy approach through the madibular area into the parapharyngeal space under the guidance of computed tomography (CT) for NPC.

METHODSBetween July 6, 2005 and October 23, 2009, a total of 6 patients were enrolled into the study. Two patients with cervical lymph node metastasis were clinically suspicious of NPC according to their clinical manifestations. However, no cancer cell could be found by repeated nasopharyngeal biopsies followed by histologic examinations. The other 4 patients were diagnosed with recurrent NPCs by magnetic resonance imaging (MRI) or/and positron emission tomography (PET)-CT scan, showing tumors in the parapharyngeal spaces in 3 patients and enlarged retropharyngeal lymph node in 1 patient. The CT-guided puncture was performed through the mandibular skin and the cutting needle biopsy was taken at the parapharyngeal space focus.

RESULTSAll the cutting needle biopsies of projected locations have been performed safely. Finally, all the 7 specimens met the requirement of pathologic diagnosis and the cases were all confirmed histologically to be NPCs. The main complication was mild ache at the puncture point. No blood vessel or nerve was injured and no patient needed special treatment.

CONCLUSIONSThe CT-guided puncture biopsy of the parapharyngeal space through the mandibular area is simple and feasible. It can be an additional option for routine nasopharyngeal biopsy.

Adult ; Aged ; Biopsy, Needle ; methods ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Magnetic Resonance Imaging ; Male ; Mandible ; Middle Aged ; Nasopharyngeal Neoplasms ; diagnosis ; diagnostic imaging ; pathology ; Neoplasm Recurrence, Local ; Pharynx ; pathology ; Positron-Emission Tomography ; Tomography, X-Ray Computed

OBJECTIVETo investigate the effects of different subtypes IFN alpha (IFN alpha2b, IFN alpha2a, and IFN alpha1b) transduction molecular STAT1, STAT2, IFNAR, PKR, and RNase L, and to study the differences of their antiviral effects and to evaluate the key signaling transduction molecules.

METHODS(1) After HepG2 cells were treated with IFN alpha2b, IFN alpha2a, or IFN alpha1b, the mRNA levels of STAT1, STAT2, IFNAR, PKR, and RNase L were detected by RT-PCR. (2) After HepG2 cells were treated with 1000 U/ml IFN alpha2b, IFN alpha2a, or IFN alpha1b, the protein expression levels of STAT1 and IFNAR were examined by Western blot.

RESULTSRT-PCR results: (1) IFNAR, STAT1, and STAT2 mRNA expression levels were slightly higher in the IFN alpha1b group than those in the IFN alpha2b group (P > 0.05). The mRNA expression levels in IFN alpha1b or IFN alpha2b groups were significantly higher than in the IFN alpha2a group (P < 0.05). (2) The PKR mRNA expression showed no significant differences among IFN alpha1b, IFN alpha2b, and IFN alpha2a groups. (3) The RNase L mRNA expression was very weak. We could not compare the differences of the RNase L mRNA levels in different groups by RT-PCR. Western blot results: (1) The IFNAR, and STAT1 protein expressions were greatly up-regulated after IFN alpha induction compared with the untreated group (P < 0.05). (2) The IFNAR, and STAT1 protein expression levels in IFN alpha1b group were slightly higher than the IFN alpha2b group. IFNAR, and STAT1 protein levels of IFN alpha1b or IFN alpha2b group were significantly higher than IFN alpha2a group (P < 0.05).

CONCLUSIONSTAT1, STAT2, IFNAR mRNA and protein expressions could all be markedly up-regulated after IFN alpha treatment. Effects of IFN alpha1b or IFN alpha2b were greatly stronger than IFN alpha2a. The PKR mRNA expression also was greatly up-regulated after IFN alpha treatment. Expression levels of PKR in IFN alpha1b, IFN alpha2b, and IFN alpha2a groups were all similar. The mRNA level results were consistent with the protein level results. Our results showed that the antiviral activity of IFN alpha1b or IFN alpha2b were stronger than that of IFN alpha2a. The signal transduction molecules STAT1, STAT2, and IFNAR could be regarded as a key index to evaluate antiviral activity of IFN alpha. Further confirmation is still needed to see whether PKR could be regarded as a key index.

Antiviral Agents ; pharmacology ; Carcinoma, Hepatocellular ; virology ; Humans ; Interferon-alpha ; pharmacology ; Liver Neoplasms ; virology ; Recombinant Proteins ; STAT1 Transcription Factor ; biosynthesis ; genetics ; STAT2 Transcription Factor ; biosynthesis ; genetics ; Signal Transduction ; Tumor Cells, Cultured