The most effective sequence of small interfering RNA (siRNA) silencing STAT3 of psoriatic keratinocytes (KCs) was screened out,and the effects of the most effective siRNA combined with ultrasonic irradiation and SonoVue microbubbles on the expression of STAT3 of KCs and the dose-and time-response were investigated.Three chemically-synthetic siRNAs targeting STAT3 carried by Lipofectamine 3000 were transfected into KCs,and the effects on STAT3 expression were detected,then the most effective siRNA was selected for the subsequent experiments.The negative controls of siRNA (siRNA-NC) labeled with Cy3 carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles were transfected into KCs,then the optimal parameters of ultrasonic irradiation were determined.The most effective siRNA carried by Lipofectamine 3000 combined with ultrasonic irradiation at the optimal parameters and SonoVue microbubbles was transfected into KCs,and the dose-and time-response of RNA interference was determined.The effect of RNA interference by the most effective siRNA at the optimal time and dose carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles (LUS group) was compared with that only carried by Lipofectamine 3000 (L group).The results showed that siRNA-3 achieved the highest silencing efficacy.0.5 W/cm2 and 30 s were selected as the parameters of ultrasonic irradiation.The siRNA-3 carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue mierobubbles could effectively knock down the STAT3 expression at mRNA and protein levels in dose-and time-dependent manners determined at 100 nmol/L with maximum downregulation on mRNA at 48 h,and on protein at 72 h after transfection.The LUS group achieved the highest silencing efficacy.It was concluded that siRNA-3 carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles could effectively knock down the STAT3 expression in psoriatic KCs,and the optimized transfection condition and the sequence of siRNA-3 could serve for further research on gene therapy of psoriasis.

Objective To evaluate the effect of gabapentin for treatment of neuropathic pain induced by bilateral sciatic nerve chronic constriction injury (bCCI) of rats.Methods Forty-eight female SPF Sprague-Dawley rats,weighing 180-200 g,aged 7-9 weeks,were randomly divided into 4 groups (n =12 each) using a random number table:control group (C group),sham operation group (S group),bCCI group,and bCCI + gabapentin group (G group).Gabapentin 100 mg/kg was intraperitoneally injected once at 15 min before operation and twice a day during 7-13 days after operation for 7 consecutive days.The bilateral mechanical paw withdrawal threshold (MWT),thermal paw withdrawal latency (TWL) and number of cold-stimulated paw withdrawal were measured before operation (baseline) and at 1,3,6,8,10,14 days after operation.Results Compared with C group,the bilateral MWT was significantly decreased,and the bilateral TWL was shortened in bCCI and G groups,and no significant change was found in MWT and TWL in S group.Compared with bCCI group,the bilateral MWT was significantly increased,the bilateral TWL was prolonged,and no significant change was found in the number of cold-stimulated paw withdrawal in G group.Conclusion Gabapentin can relieve thermal and mechanical hyperalgesia induced by bCCI,however,it exerts no effect on cold allodynia in rats.

OBJECTIVETo develop a HPLC-ESI-MS method for the determination of puerarin and its metabolite and study the metabolic kinetics in beagle dog liver microsomes.

METHODBeagle dog liver microsomes were prepared by using ultracentrifugation method. Chromatography was performed on a Shimadzu C18 column (2.0 mm x 150 mm, 5 microm). Amethanol-water gradient system was used. ESI interface was applied in the positive, and SIM m/z 417 was puerarin and m/z 531 was daidzein.

RESULTThe puerarin was metabolized by NADPH regenerating system in beagle dog microsomes. The Michaelis-Menten parameters Km and Vmax in beagle dog microsomes were initially estimated by analyzing Lineweave-Brurk plot. The Vmax Km of puerarin were (0.047 +/- 0.006) mg x min(-1) x g(-1), (1.22 +/- 0.53) mg x L(-1).

CONCLUSIONThe puerarin and daidzein can be rapidly determined by HPLC-MS in beagle dog microsomes and the puerarin was metabolized to daidzein by CY P450. The study can give help for Baige capsule.

Animals ; Chromatography, High Pressure Liquid ; Dogs ; Isoflavones ; pharmacokinetics ; Liver ; chemistry ; drug effects ; Microsomes, Liver ; chemistry ; drug effects ; Pharmacokinetics ; Spectrometry, Mass, Electrospray Ionization

Adult ; Dizziness ; prevention & control ; Humans ; Hypnotics and Sedatives ; pharmacology ; Male ; Midazolam ; pharmacology ; Naval Medicine ; Young Adult

OBJECTIVETo explore the relationship between methylenetetrahydrofolate reductase (MTHFR) gene polymorphism and microsatellite instability (MSI) in patients with gastric cancer.

METHODSMTHFR gene C677T and A1298C polymorphism were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and MSI was examined with PCR.

RESULTSMTHFR gene C677T and A1298C polymorphisms were analyzed on 122 gastric cancers and 110 normal controls The genotype frequencies of MTHFR 677CC, 677CT and 677TT were 47.5%, 39.3% and 13.1% on patients with gastric cancer, and 48.5%, 42.6%, 8.9% in the controls respectively. There was no significant difference of genotype frequency between the two groups (P > 0.05). The individuals with 677CT genotype, 677TT genotype and 677CT + TT genotype exhibited significantly reduced risk (OR = 0.38,95% CI: 0.15-0.98; OR = 0.26,95% CI: 0.03-2.18 and OR = 0.36,95% CI: 0.07-0.98) of developing gastric cardia cancer compared with those harboring the wild-type(677CC). The individuals with 677TT genotype having a 3.03-fold (95% CI: 1.07-8.65) increased risk of developing gastric corpus cancer. The genotype frequency of MTHFR 1298AA, 1298AC and 1298CC were 59.8%, 36.1% and 4.1% in gastric cancer patients, and 57.4%, 7.6%, 5.0% in the controls, respectively. The distribution of MTHFR A1298C genotype was not significantly different between gastric cancer and controls (P > 0.05). The individuals with 1298CC genotype had a reduced risk of developing gastric antrum cancer (OR = 0.41- fold, 95% CI: 0.03-2.18, 0.05-3.72) when comparing with those having 1298AA genotype. Patients with MSI+ gastric cancer had an increased frequency of the MTHFR 677TT genotype when comparing with those suffering from MSI- gastric cancer (P = 0.009) and with controlled subjects (P = 0.008). There was no significant association found between MTHFR A1298C polymorphism and MSI (P>0.05).

CONCLUSIONPolymorphism of MTHFR C677T was associated with increased risk on gastric corpus cancer and reduced risk on gastric cardia cancer. The polymorphism of MTHFR A1298C was associated with reduced risk for gastric antrum cancer while MSI pathway was possibly involved in the development of gastric cancer with MTHFR 677TT genotype.

Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Microsatellite Instability ; Middle Aged ; Polymorphism, Genetic ; Stomach Neoplasms ; genetics

This study purposed to investigate the effects of different oxygen concentrations and reactive oxygen species (ROS) on the biological characteristics of hematopoietic stem cells (HSC) and their possible mechanisms through simulating oxygen environment to which the peripheral blood HSC are subjected in peripheral blood HSCT. The proliferation ability, cell cycle, directed differentiation ability, ROS level and hematopoietic reconstitution ability of Lin(-)c-kit(+)Sca-1(+) BMHSC were detected by using in vitro amplification test, directional differentiation test, cell cycle analysis, ROS assay and transplantation of Lin(-)c-kit(+)Sca-1(+) HSC from sublethally irradiated mice respectively. The results showed that oxygen concentrations lower than normal oxygen concentration, especially in hypoxic oxygen environment, could reduce ROS generation and amplify more primitive CD34(+)AC133(+) HSC and active CD34(+) HSC, and maintain more stem cells in the G(0)/G(1) phase, which is more helpful to the growth of CFU-S and viability of mice. At the same time, BMHSC exposed to normal oxygen level or inconstant and greatly changed oxygen concentrations could produce a high level of ROS, and the above-mentioned features and functional indicators are relatively low. It is concluded that ROS levels of HSC in BMHSCT are closely related with the oxygen concentration surrounding the cells and its stability. Low oxygen concentration and antioxidant intervention are helpful to transplantation of BMHSC.
Animals ; Bone Marrow Cells ; cytology ; metabolism ; Cell Differentiation ; Cells, Cultured ; Female ; Hematopoietic Stem Cells ; cytology ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Oxygen ; administration & dosage ; pharmacology ; Reactive Oxygen Species ; metabolism

Objective To determine whether morphine having the ability to influence the antiviral effect of lamivudine(3TC)in vitro study.Methods MT2 cells were randomly assigned into morphine+3TC treatment group,morphine+naloxone+3TC treatment group,naloxone+3TC treatment group.Both 3TC and virus control groups were set up.The corresponding MT2 cells were treated with opiates antagonist(naloxone)for 0.5 hours before the 24-hours morphine treatment program was implemented while all of the groups were then infected with equal amounts of cell-free HIV-1 ⅢB strain and 3TC.HIV-1 p24 antigen in culture supernatants collected at days 3,4,5 and 6after infection status was tested and the inhibition of 3TC anti-HIV-1 p24 antigen of various treatment groups calculated.Results Inhibition of 3TC anti-HIV-1 p24 antigen of Morphine+3TC treatment group was the lowest when HIV-1 infected cells at 3rd and 4th day and showed significant difierence (P<0.05)when compared to the 3TC control.However,there was no statistically significant difference among them(P>0.05),when virus was infected the cells at 5th and 6th day.The difference of 3TC anti-HIV-1 p24 antigen inhibition between the morphine+naloxone+3TC treatment group and the naloxone+3TC treatment group was not significant(P>0.05).Similar results were obtained when these two groups were compared to the 3TC control group(P>0.05),respectively.The 3TC anti-HIV-1 p24 antigen inhibition of each treatment group reduced as the time of infection prolonged,showing a significant and time-course effbct.Conclusion The 3TC antiviral effect was reduced by morphine in the early stage of infection,and could be blocked by naloxone.

OBJECTIVETo study the antibiotic- and disinfectant-resistance features of and disinfectant-resistant gene distribution in Staphylococcus aureus (Sa) isolated from the urogenital tract of male patients with urogenital tract infection (UTI). total of 152 Sa isolates were collected from the urethral discharge specimens from male UTI patients. The minimum inhibition concentration (MIC) of antimicrobial agents and disinfectants commonly used against Sa were tested by standard ager dilution; the methicillin-resistant Sa (MRSA) isolates detected by cefoxitin disk diffusion and mecA gene amplification; Staphylococcal cassette chromosome mec (SCCmec) genotyping performed by multiplex PCR; the disinfectants gene qac (quaternary ammonium compound) amplified by PCR; and the clonal relatedness of qacA/B-positive MRSA isolates investigated by pulsed-field gel electrophoresis (PFGE).

RESULTSOut of the 152 Sa isolates, 91 (59.9%) were found to be MRSA. SCCmec genotyping showed SCCmec V to be the main type, accounting for 63.7% (58/91), with 8 (8.8%) isolates of SCCmec I, 2 (2.2%) isolates of SCCmec II, 19 (20.9%) isolates of SCCmec III, and 4 (4. 4%) isolates of SCCmec IV. The Sa isolates exhibited high rates of non-susceptibility to penicillin (95.4%) , erythromycin (72.4% ) , ciprofloxacin (42. 8%), and levofloxacin (44.7%), and a fairly high sensitivity to nitrofurantoin, teicoplanin, linezolid, and vancomycin. The MIC in the Sa isolates was 0. 25 -16 microg/ml for chlorhexidine; MIC50 and MIC90 were 2.0 and 4.0 microg/ml respectively for MRSA strains and both 1.0 microg/ml for MSSA strains. Out of the 152 Sa isolates, 72 (47.4%) harbored the qacA/B gene, 6 (3.9%) the smar (qacC + qacD) gene, 9 (5.9%) the qacE delta 1 gene, and 2 (1.3%) the qacH gene, but no qacG and qacJ genes were detected. PFGE analysis showed that the qacA/B-positive MRSA isolates were distributed

CONCLUSIONClinical Sa isolates exhibited varied degrees of resistance to commonly used antibiotics, and in a polyclonal manner. some showed a robust tolerance to chlorhexidine. The main disinfectant-resistant gene is qacA/B. Antimicrobial agents and disinfectants should be used rationally according to clinicians.

Disinfectants ; pharmacology ; Drug Resistance, Bacterial ; genetics ; Genotype ; Humans ; Male ; Staphylococcus aureus ; drug effects ; genetics ; Urinary Tract Infections ; microbiology

OBJECTIVETo observe the clinical effects of the no-flip procedure with the Chinese Shang Ring when circumcising adult males with redundant prepuce or phimosis, and to discuss its advantages and disadvantages.

METHODSUsing the no-flip Shang Ring technique, we performed circumcision for 167 adult males aged 18 -72 (mean 27.8) years with redundant prepuce or phimosis, and analyzed the clinical data, including the operation time, postoperative complications, ring-removal time, and postoperative appearance of the penis.

RESULTSComplete follow-up data of 94 cases (56.29%) were obtained. The mean operation time was (5.03 +/- 0.71) minutes and the average ring-removal time was (18.83 +/- 6.70) days. The primary postoperative complications were edema (35 cases [37.23%] at 2 weeks and 9 cases [9.57%] at 4 weeks), including 2 severe cases (2.13%), and infection (3 cases [3.19%]). The pain scores were 2.01 +/- 2.46 during the procedure and 4.52 +/- 2.53 at 24 hours postoperatively. Slipping of the outer ring occurred in 1 case, and delayed removal of the ring in 30 cases (31.91%).

CONCLUSIONAdult male circumcision with the no-flip Shang Ring technique is recommended for its short operation time, simple procedure, fewer postoperative complications, less pain, and better incision appearance.

Adult ; Aged ; Circumcision, Male ; adverse effects ; instrumentation ; methods ; Edema ; etiology ; Humans ; Male ; Middle Aged ; Operative Time ; Pain, Postoperative ; etiology ; Penile Diseases ; etiology ; Penis ; abnormalities ; surgery ; Phimosis ; surgery ; Postoperative Complications ; Postoperative Period ; Prostheses and Implants ; Young Adult

Objective To investigate the effects of sodium nitroprussiate on neurological function and Janus family of tyrosine kinases/signal transduction and transcription activator 3 (JAK2/STAT3) pathway in rats with experimental intracerebral hemorrhage. Methods Sixty rats were randomly divided into control group,model group and test group,20 rats in each group. The model in model group and test group were given collagenase and heparin mixture to establish experimental cerebral hemorrhage,while control group was given the same amount of 0. 9％ NaCl. The test group received intraperitoneal injection with sodium nitroprussiate(1 mg·kg-1) at 6,24,72 and 120 h after modeling,and the rats in model group and control group received intraperitoneal injection with the same amount of 0. 9％ NaCl. The brain water content of each group was measured by weighing method was calculated; TdT-mediated dUTP nick- end labeling (TUNEL) method was used to detect apoptosis of rat brain tissue,and Western blotting (WB) method was used to detect the expression of JAK2/STAT3 pathway-related protein in rat brain tissue. Results At 72 h after modeling,there were significant differences among the control group,model group and test group in the brain water content [(76. 05 ±2. 04) ％ vs (84. 91 ±2. 58) ％ vs (79. 68 ±3. 01) ％],the rate of apoptotic neurons [(7. 52 ± 1. 86) ％ vs (50. 23 ± 2. 43) ％ vs (26. 24 ± 2. 37) ％],the relative expression of p- JAK2 protein in the brain tissues [(0. 07 ± 0. 13) vs (1. 26 ± 0. 05) vs (0. 33 ± 0. 05) ]and the relative expression of p-STAT3 protein [(0. 20 ± 0. 03) vs (0. 38 ± 0. 06) vs (0. 26 ± 0. 09) ](all P < 0. 05). Conclusion Sodium nitroprussiate has protective effect on brain injury in experimental cerebral hemorrhage rats,which may be related to regulate JAK2/STAT3 pathway and reduce apoptosis of brain tissue.