Objective To establish a rapid and accurate method of fluorescent quantitative PCR detection for Avian Influenza Virus H7N9 type.Methods A pair of primers and TaqMan fluorescent probes were designed specificly for H7N9 gene ac-cording to the published nucleotide sequence from National Center for Biotechnology Information.Positive recombinant plas-mid was built and the minimum copies of detection,repeatability and specificitv were tested.Results The results showed that this assay obtained positive recombinant plasmid,the linear relation was fine in the range from 102 to 108 copies/μl.The minimum copies was 500 copies/μl with this method,specificitv and repeatability(CV<1%)were fine,no cross reaction with other viruses.The precision was 100% (16/16)with this method for detecting 8 positive samples and 8 negative sam-ples.Conclution The establishment methods of fluorescent quantitative PCR detection could be used for the rapid diagnosis Avian Influenza virus H7N9 types.

Objective To investigate the preparation process of cold -dampness medicine iron stick and the curative effect of cold -dampness medicine iron stick,thus to provide reference for the clinical treatment of cold -dampness lumbago.Methods Using science and technology of preparation and the thermal effect of cold -dampness medicine iron stick was observed.1 20 cases with cold -dampness lumbago of dampness type with low back pain, according to the order of treatment were divided into the observation group and control group.63 cases of the treatment group were given cold -dampness medicine iron stick affixed to the surface of the skin,57 cases of the control group were given kanlisha affixed to the surface of the skin.The treatment effect after one course of treatment was observed. Results Cold -dampness medicine iron stick heat effect index of heat time was 1 2min,to thermal time was 1 0.8h, the highest temperature was 56.5℃,thermal equilibrium curve was gentle and lasting.The efficacy of the two groups of drugs within the observation time effect was obvious.The total effective rate of the observation group was 90.5%(57 /63),which was higher than 75.4%(43 /57)of the control group,the difference was statistically significant (χ2 =4.87,P <0.05).During the treatment,local skin blisters,rash,itching and other low temperature burns and other allergic reaction were not found.Conclusion Cold -dampness medicine iron stick process is stable and controllable in quality and heat balance curve slow long -lasting,determine the efficacy,innovative therapies,it is easily accepted by patients and worthy to be popularized in clinical practice.

Objective To assess the efficacy and safety of transurethral resection of the prostate(PKRP) and bladder tumor(PKRBT) with plasmakinetic energy. Methods 300 patients(mean age of 71.6 years;range from 52 to 91) with symptomatic BPH without suspected cancer (confirmed by IPSS,urinary flow rate,ultrasonographic estimate and PSA tests) were treated by PKRP. 37 patients with superficial transitional cell carcinoma (17 with tumors appearing at the lateral wall) were treated by PKRBT.All patients were followed up for 1～6 months postoperatively. Results For PKRP,the duration of the procedure was (48?31) min(range 13 to 98 min).No case need blood transfusion during the operation.No transurethral resection syndrom occurred.The mean catheterization time was 2 days (1 to 4 days). The peak flow rate increased from (5.2?3.9) to (19.2?4.1),(21.8?4.6),(22.3?5.7)ml/s at 1,3 and 6 months respectively. The IPSS decreased from 24.7 to 5.7, 5.4,5.0 and the QOL decreased from 5.3 to 1.7,1.8,1.2 at 1,3 and 6 months respectively.For PKRBT,the duration of the procedure was (24?13) min(range 5 to 39 min) with little bleeding.The adductor reflex was found in 5 of the 17 cases with tumor located at the lateral wall. Conclusions It is suggested that transurethral bipolar plasmakinetic resection of the prostate and bladder tumor is effective and safe. This pilot series permits a comparative study with conventional TUR system.

[Objective] To search for a new technique for incarcerated annular mixed hemorrhoids (IAMH) . [Methods] One hundred and eighteen cases of IAMH (group A) were treated by double denenate excision-ligation combined with pad preservation and 53 eases (group B) by external-exfoliation and internal-ligation. Cure rate, wound-healing time, anal function and postoperative complications were observed in the two groups. [Results] Cure rate was 96.6% in group A and 84.9% in group B; wound-healing time was (17 ? 5) days in group A and (24 ? 5) days in group B ( P

Objective To assess the efficacy and safety of transurethral bipolar electrocautery resection of the superficial bladder tumor(Bipolar-TURBt).Methods 26 patients with superficial transitional cell carcinoma(13 with tumors appear at the lateral wall) were treated by bipolar TURBt. All patients were followed-up in 1～6 months postoperatively.Results The duration of the procedure was(19?10) min(8～35 min) with little bleeding. Bladder perforation was occurred in no case. No TUR-syndrom was found. The adductor reflex was not found even in the 13 cases with tumor located at the lateral wall.Conclusion Transurethral bipolar electrocautery resection of the superficial bladder tumor is effective and safe.

Objective To investigate the methods of prevention and treatment of presacral venous plexus bleeding in pelvic operation.Methods The clinical data of 8 cases of presacral venous plexus bleeding in pelvic operation from 1998 to 2013 were analyzed.Results All 8 cases succeeded in controlling bleeding,The amount of bleeding was 1 000-4 000 mL,the average amount is 2 600 mL.Conclusions The key to prevention of presacral venous plexus bleeding was thorough familiar with pelvic anatomy and a dexterous technique of careful dissection.Massive hemorrhage occurred direct electric coagulation hemostasis was available,the method was simple.

BACKGROUND: Human umbilical cord Wharton's jelly-derived mesenchymal stem cells (hWJCs) via transplantation are able to survive in a dog model of intervertebral disc degeneration and to partially restore the height of the intervertebral disc. However, there is no study on whether hWJCs can repair nucleus pulposus cells after inflammatory injury as yet. OBJECTIVE: To observe the protective effect of hWJCs on interleukin-1β induced rat nucleus pulposus degeneration.METHODS: Nucleus pulposus cells at passage 3 from Sprague-Dawley rats were divided into three groups: (1) normal control group cultured in culture medium containing fetal bovine serum for 24 hours, and then cultured in complete medium for 24 hours; (2) intervention group with interleukin-1β intervention for 24 hours, followed by culture in the complete medium for 24 hours; (3) co-culture group with interleukin-1β intervention for 24 hours, followed by culture in the complete medium for non-direct co-culture with hWJCs for 24 hours. Annexin V-FITC/propidium iodide (PI) double staining and flow cytometry were used in succession to measure the apoptosis rate of nucleus pulposus cells in each group. The ADAMTS-4, MMPs-3 and TIMP-1 expression in nucleus pulposus cells was observed though RT-PCR. The activities of caspase-3, caspase-8 and caspase-9 were determined by caspase kit.RESULTS AND CONCLUSION: The apoptosis rates were ranked as follows: the intervention group > co-culture group > normal control group, and there were significant differences between groups (P < 0.05). Compared with the control group,the expression levels of ADAMTS-4 and MMPs-3 were significantly higher and the expression level of TIMP-1 was significantly lower in the intervention and co-culture groups (P < 0.05). Compared with the intervention group, the expression levels of ADAMTS-4 and MMPs-3 were significantly lower and the expression level of TIMP-1 was significantly higher in the co-culture group (P < 0.05). The expression activities of caspase-3, caspase-8 and caspase-9 were highest in the intervention group, followed by the co-culture group, and lowest in the normal control group, and there were significant differences between groups (P < 0.05). To conclude, hWJCs has obviously inhibitory effect on the apoptosis of degenerated nucleus pulposus cells induced by interleukin-1β.

An HPLC method for the determination of 18alpha-glycyrrhetinic acid and 18beta-glycyrrhetinic acid in rat plasma was established, which was used subsequently to determine the pharmacokinetic profiles of both epimers of glycyrrhetinic acid in rats. alpha-glycyrrhetinic acid, beta-glycyrrhetinic acid, and a mixture of alpha-glycyrrhetinic and beta-glycyrrhetinic acids were administered to rats via gastric infusion. Blood samples were collected at different time intervals and extracted by liquid-liquid extraction. Separation was achieved by using a Kromasil C18 column (150 mm x 4.6 mm, 5 microm) with the mobile phase composed of acetonitrile--4 mmol x L(-1) ammonium acetate solution (46 : 54, v/v) at a flow rate of 1.0 mL x min(-1), and the detection wavelength was set at 250 nm. The pharmacokinetic parameters were calculated using the software DAS 2.0. In a combined administration, the main pharmacokinetic parameters of beta-glycyrrhetinic acid are significantly different from that of alpha-glycyrrhetinic acid (P < 0.05), while no significant difference was obtained when administrated individually. Compared to the single administration, significant differences (P < 0.05) on the values of AUC(0-t) and AUC(0-infinity) of beta-glycyrrhetinic acid were observed when this chemical was administrated together with alpha-glycyrrhetinic acid. In contrast, the pharmacokinetic parameters of alpha-glycyrrhetinic acid were not affected even under the co-administration. Here, a sensitive, specific, rapid and reproducible HPLC method was developed for the pharmacokinetic studies of alpha-glycyrrhetinic acid and beta-glycyrrhetinic acid in rat plasma.

Digital hospital construction is foundational project for the modern management,which challenges the management mode and idea in hospital.With regard to the status on the construction of digital hospital of infectious disease in 302 hospital of the People's Liberation Army,The article reviews the experience and prospect of digital hospital of infectious disease.

Objective To investigate the role of preoperative peripheral blood neutrophil to lymphocyte ratio (NLR),platelet to lymphocyte ratio (PLR),prognostic nutritional index (PNI) in the prognosis of patients with hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) after radical operation.Methods This is a retrospective study,involving 426 surgically resected hepatitis B related hepatocellular carcinoma cases in a single center from 2003 to 2012.Results Kaplan-Meier analysis showed patients in NLR ≤ 1.62 group achieve higher rate of recurrence-free and overall survival than that in the NLR ＞ 1.62 group,the difference was statistically significant (P ＜ 0.005);Also PNI ＞ 49.42 group showed higher rate of overall survival significantly than PNI≤49.42 group (P ＜ 0.005).The results of Cox regression multivariate analysis further suggested that both NLR ＞ 1.62 (HR 1.74,P =0.007) and PNI ≤49.42 (HR 0.70,P =0.021) were independent risk factors for overall survival,NLR ＞ 1.62 (HR 1.45,P =0.03) was also an independent risk factor for recurrence-free survival.Conclusion The preoperative NLR and PNI may be independent risk factors for prognosis of patients with HBV-related HCC after radical operation.