Objective To construct the autocatalytic caspase-3 and investigate its apoptosis- inducing effect in ovarian cancer in vitro and in vivo.Methods PCR recombination technique was used to construct autocatalytic caspase-3 which is named as rev-caspase-3,and Ad-Max system was used to prepare recombinant adenovirus containing rev-caspase-3,which is named as Ad-rev-easp3.Immunohistochemistry was used to detect active caspase-3 expression.Cell counting kit,flow cytometry and western blot were used to measure cell survival rate,apoptotic rate,cell cycle distribution and the expressions of plT,active subunit of caspase-3,and p85,the poly(adenosine diphosphate-ribose)polymerase(PARP)cleavage segment,respectively.Transmission electron microscope was used to detect cell ultrastrueture,and real time PCR was used to detect apoptosis-related gene expression.Subcutaneous tumor models and abdominally spread tumor models of human ovarian carcinoma were established using AO cells in BALB/c nude mice. The mouse survival rates were measured for abdominally spread tumor models,and the volume of tumor nodules were determined for subcutaneous tumor models following the treatments of rev-caspase-3.Results Active caspase-3 protein was significantly expressed,and the expression levels of active subunit of caspase- 3,p17,and the PARP cleavage segment,p85,were significantly elevated in cells treated with rev-caspase- 3.The decrease of cell survival rate and the increase of cell apoptotic rate were detected following Ad-rev- casp3 treatment.Treatments with Ad-rev-casp3 [ multiplicity of infection(MOI)was 70 ] resulted in survival rate of 30.3% and apoptotic rate of 40.2%.There was a significant increase in cell number of S- phase(56.5%),while there was no significant apoptosis(3.4%)following treatments with Ad-rev-casp3 at a low dosage of MOI=10.Cells treated with rev-caspase-3 displayed significant apoptotic morphology. The levels of active caspase-3 gene expressions(9.44)significantly increased.Rev-caspase-3 treatment significantly prolonged survival,the mean survival duration was(213?16)days,and suppressed tumor growth(tumor growth suppression rate was 70%),when compared with treatment with phosphate buffered saline(PBS).Conclusion Recombinant adenovirus containing rev-caspase-3 can significantly induce apoptosis of ovarian carcinoma cells,suppress tumor growth and prolong the mouse survival duration.

Objective To study the flurbiprofen axetil (FA) for postoperative analgesia of gynecological laparoscopic surgery and the changes of interleukin-6 in blood.Methods80 cases were selected for gynecologic laparoscopic surgery,the patients were randomly divided into 4 groups with 20 cases each group.Group A was given FA 50 mg and tramadol 1 mg/kg 30 minutes before the end of surgery.Group B was given FA 50 mg 5 minutes before induction of anesthesia and given tramadol 1 mg/kg 30 minutes before the end of surgery,and group C was given FA 50 mg five minutes before induction of anesthesia.Group D was given tramadol 2 mg/kg 30 minutes before the end of surgery.After the surgery,the VAS was observed in real-time of wake up (T1),and 2 hours (T2),6 hours (T6) and 24 hours (T24) after operating.The blood of each group was taken at the different time points including arriving in the room (T0),waking up (T1) and 6 hours (T6) after operating.The blood samples were hold for 30 minutes and centrifuged (3000 r/min,10 min),and IL-6 was detected using double-antibody sandwich ABC-ELISA.ResultsThe experiments showed that VAS in T1 time point of group A,B,C were lower than that of group D(1.40±0.26,1.67±0.37,1.60±0.42 vs 3.13±2.32).In T24 time point,VAS of group A,B were also lower than that of group D(2.13±1.24,2.00±1.25 vs 3.53±1.87),the differences were significant (P＜0.05).The experiments showed that concentration of IL-6 in group D was higher than that in group A,B,C at the T1 time point (12.26±6.56 vs 2.94±2.55,3.37±2.43 vs 2.93±1.16,P＜0.05).ConclusionsThe postoperative analgesic effect of Flurbiprofen axetil combined with tramadol on the ogynecological laparoscopic was superior to using any of the drug alone.Flurbiprofen axetil could inhibit increasing of IL-6 and have an effect against the inflammatory response induced by surgical trauma,and it has a good analgesic effect.

OBJECTIVE To investigate the disinfection method for indwelling urinary catheter and its correlation with the urinary infection.METHODS Eighty cases with indwelling catheter were randomly assigned to two groups.0.5% Polyvidone iodine(Iodophor)was used as lubricant and disinfectant in the experimental group.The control group received liquid paraffin and 0.1% benzalkonium bromide.RESULTS The incidence of urinary tract infections in the experiment group was significantly lower than that in the control group with the same indwelling time of catheter(P

Objective To investigate the effect of intermittent low flow oxygen inhalation on cognitive function in patients with vascular dementia. Methods 116 cases of vascular dementia were divided into treatment group (n=58) and control group (n=58). Both groups received the routine drugs, and the treatment group received intermittent low flow oxygen inhalation in addition. They were assessed with the Mini Mental Statement Examination (MMSE) and Activity of Daily Living Scale (ADL) before and 8, 16 weeks after treatment. Results There was not significant difference between groups in scores of MMSE and ADL 8 weeks after treatment, but was 16 weeks after treatment (P<0.01). Conclusion Intermittent low flow oxygen inhalation can improve the cognitive function and quality of life in patients with vascular dementia.

Objective To investigate the antitumor effect of flavopiridol in ovarian cancer. Methods After the treatment with flavopiridol of AO cells,cell apoptotic rate and cell cycle distribution were detected by flow eytometer and the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labelling(TUNEL).Real time PCR was used to detect the expression of cyclin D and active caspase-3 in AO cells.Subcutaneous tumor models and abdominally spread tumor models of human ovarian carcinoma using AO ceils in BALB/c nude mice were established.The mouse survival rates were measured for abdominally spread tumor models and the volume of tumor nodules was determined for subcutaneous tumor models following the treatments of flavopiridol.TUNEL was used to detect cell apoptosis,and immunohistochemistry was used to measure microvessel density(MVD)in tumor tissues. Results AO cells showed apoptotic rates of 4.1%,10.7% and 7.6% following the treatments with flavopiridol at 150,300 and 500 nmol/L respectively,accompanied by an increase in G_1 progression and a decrease in S phase progression.The level of active caspase-3 increased(2.55 vs 2.49)and the level of cyclin D expression decreased significantly(0.25 vs 0.69,P

Objective To investigate the antitumor effects on ovarian cancer using recombinant adenoviruses expressing autocatalytic caspase-3 driven by amplified human telomerase reverse transcriptase promoter (AdHTVP2G5-rev-casp3) combined with flavopiridol. Methods Following the treatment with AdHTVP2G5-rev-casp3 combined with flavopiridol, cell survival rate was measured by cell counting kit 8;cell apoptotic rate and cell cycle distribution were detected by flow cytometry. Western blot was performed to observe the expression of p17, the active subunit of caspase-3, and p85, the cleavage segment of substrate of caspase-3, in AO cells. The mice survival rates were measured for abdominally metastatic tumor models and the volume of tumor nodules were determined for subcutaneous tumor models following the treatments of AdHTVP2G5-rev-casp3 combined with flavopiridol. HE staining was used to detect the histopathological changes of various organs, and the serum level of alanine transaminase (ALT) and aspartate aminotransferase (AST) were measured to monitor liver damages following the intraperitoneal administration of AdHTVP2G5-rev-casp3 and flavopiridol. Results There was no significant cell-killing effects or apoptosis in AO cells following treatments with AdHTVP2G5-rev-casp3 or flavopiridol at low dosage alone (apoptotic rate all ＜ 11% ), whereas significant synergism of their sequential combination was observed in AO cells. This sequential treatment of AdHTVP2G5-rev-casp3 [multiplicity of infection (MOI) was 20]infection for 72 hours, followed by flavopiridol ( 300 nmol/L) for 48 hours, could result in the most substantial cell death, and AO cells survival rate and apoptotic rate were 73. 5% and 11.6%, respectively.Following treatments with AdHTVP2G5-rev-casp3 at low doses ( MOI = 10), there was a significant increase in cell number with S-phase content ( 62. 5% ), which resulted in the most marked apoptosis induced by sequential treatments with flavopiridol. The sequential combination could induce significantly higher levels of p17 and p85 expression than that when their applications alone. Combined AdHTVP2G5-rev-casp3 and flavopiridol treatment prolonged mouse survival [ mean survival time of ( 286 ± 6) days ] and suppressed tumor growth significantly (tumor growth suppression rate of 81% ), when compared with treatment using either alone. The levels of serum ALT and AST were not significantly elevated and no obvious lesions were found in any organs in treatments with AdHTVP2G5-rev-casp3 of low doses combined with flavopiridol.Conclusions AdHTVP2G5-rev-casp3 at low doses results in a significant increase in cell number with Sphase content, which significantly enhanced the sensitivity of cells to flavopiridol. Treatments of autocatalytic caspase-3 combined at low doses with flavopiridol result in significant synergistic antitumor effects,significant tumor growth suppression and prolonged survival of mice. When compared with normal dose flavopiridol alone, the combination could resulted in minimal liver toxicity.

With the establishment of HPLC and LC-MS methods to determine the related substances and the content of active pharmaceutical ingredient (API) in ipratropium bromide aerosol products, several packing material-related impurities were identified, including antioxygen BHT and antioxygen 2246. Results showed that these leachable additives from the packing materials may present at a relative high level in the drug solution, and the low content of API in the drug products is usually due to the adsorption of the packing material as well as the leaking of contents. The current available assay methods for the control of ipratropium bromide aerosol products are often lack of specificity and unable to assure the drug quality effectively. To meet the increasing attention on the regulations of drug packing materials, our research would be a pilot study, indicating that the inappropriate packing materials could cause the migration and adsorption of the active ingredients, and the importance to have compatibility studies between packing materials and drugs.

BACKGROUND:Guided tissue regeneration and bone grafting are a hot spot in the treatment of periodontal bone defect caused by severe periodontitis, but a smal sample size in clinical research wil lead to bias. OBJECTIVE:To systematical y evaluate the effect of guided tissue regeneration combined with bone grafting in the treatment of periodontal bone defect, and explore its feasibility, thus providing evidence for clinical application. METHODS:A computer-based search of PubMed, Cochrane Library, EMbase, CNKI, CqVip and WanFang databases was performed for articles about the guided tissue regeneration and bone graft for periodontal bone defects, published from 2000 to 2016. The keywords were“guided tissue regeneration, bone grafts, periodontal bone defects”in English and Chinese, respectively. The literature selection, data col ection and evaluation of bias were conducted by two researchers independently, and then quality assessment of the included 12 randomized control ed tests was conducted, fol owed by Meta-analysis using Revman 5.3 software. RESULTS AND CONCLUSION:A total of 12 studies were enrol ed, including 414 teeth (228 of which in the experimental group and 216 in the control group). Meta-analysis results showed that compared with the single flap surgery, guided tissue regeneration combined with bone graft could reduce periodontal probing depth by 1.18 mm gingival, make a gingival recession by 0.23 mm, reduce alveolar bone defect depth by 1.57 mm, and increase clinical attachment level by 2.03 mm (P<0.05). Compared with guided tissue regeneration technique, guided tissue regeneration combined with bone graft made probing depth increase by 0.34 mm, alveolar bone defect depth reduce by 0.73 mm, gingival recession reduce 0.35 mm and clinical attachment level increase by 0.63 mm (P<0.05). Compared with bone graft, guided tissue regeneration combined with bone graft made periodontal probing depth reduce by 0.11 mm, clinical attachment levels increase by 0.04 mm and gingival recession increase by 0.13 mm (P>0.05). These results reveal that for moderate to severe chronic periodontitis with periodontal bone defects, guided tissue regeneration combined with bone graft has better clinical effects than simply flap surgery and guided tissue regeneration, but has no significant differences from the bone graft surgery. Herein, we have not yet classified the membrane materials, bone materials and bone substitutes, and there is stil a lack of high-quality and large-sample randomized control ed trials.

Objective To elucidate the MRI appearance of prostatic abscess,the DWI and enhanced MRI features.Methods 12 cases of prostatic abscesses were retrospectively analyzed,the clinical symptom mainly manifested as lower urinary tract symptoms and fever.All of the patients were given routine MR examination including DWI sequence,6 patients received further enhanced MR examination.Results In the 12 cases,there were 4 cases behaved as single type,8 cases as multifocal type.The abscess showed iso-or slightly hypo-signal intensity on T1 WI,hyper-signal intensity on T2 WI,markedly high signal intensity on DWI and correspond-ing markedly low signal intensity on ADC.Complete abscess walls showed iso-or slightly hyper signal on T1 WI,hypo-signal inten-sity on T2 WI.The mature abscess walls were thin and smooth,which showed homogeneously ring enhanced in 4 cases.The imma-ture abscess walls showed uneven thickness and moderately enhanced in 2 cases.Septum in the abscess could be found in 4 cases, which showed similar enhancement to the abscess walls,while the abscess cavity showed non-enhanced.Abscesses involved the sur-rounding structures in 2 cases,the involved area showed obvious hyper-signal on T2 WI fat-suppression sequence.Conclusion DWI is the best sequence in the diagnosis of prostatic abscess,the markedly high signal intensity on DWI is the characteristic sign.The enhanced MRI showed the walls and septa clearly,the extent and involvement of adjacent structures.The multi-parametric MRI is a prominent procedure in the diagnosis of prostatic abcess.

Objective To explore the diagnostic value of diffusion weighted imaging (DWI) and CT perfusion imaging (CTPI) in diagnosis of liver fibrosis. Methods Fifty-seven hepatic fibrosis patients and 23 normal controls received DWI (b=500 s/mm2), ADC value of different fibrosis stages was measured, while 35 patients and all 23 normal controls received CTPI. The parameters of CTPI including blood flow (BF), blood volume (BV), mean transit time (MTT), hepatic arterial fraction (HAF) and permeability surface (PS) were measured. Analysis of variances was performed to compare the difference among the groups in both examinations. ROC curve was used to analyze the sensitivity and specificity of DWI and CTPI. Results The difference of ADC value between control group and S1 group was not significant, but between control group and S2, S3, S4 group and among group S2, group S3, group S4 was significant. In the parameters of CTPI, only the difference of HAF between control group and S3-S4 group was significant. Sensitivity and specificity of DWI and CTPI was 78.90%, 82.60% and 66.67%, 73.91%, respectively. Conclusion DWI is superior to CTPI for early diagnosis and degrading of liver fibrosis.