Objective To assess the effect of dilute pituitrin solution (0.05 U/ml) on blood loss and distention fluid intravasation during hysteroscopy. Methods From January 2003 to June 2004, 68 women with abnormal uterine hemorrhage undergoing hysteroscopic endometrial resection were randomly divided into treatment and control groups (34 cases in each). In the treatment group, dilute pituitrin solution (0.05 U/ml) was injected into the cervix before dilation of the cervix in preparation for hysteroscopy, while the patients in the control received no pituitrin injection. Results The mean operation time in the treatment group was significantly shorter than that in the control [(32.2?6.0) min vs (35.9?6.8) min; t=-2.379, P=0.020]. The amount of distention fluid infusion in the treatment group was significantly less than that of the control [(2982.1?880.5) ml vs (3461.2?795.8) ml; t=-2.354, P=0.022]. The volume of distention fluid intravasation in the treatment group was significantly less than that of the control [(225.3?81.1) ml vs (319.4?89.2) ml; t=-4.551, P=0.000]. The rate of fluid intravasation was (7.5?1.1)% in the treatment group that was significantly lower than that in the control [(9.2?1.1)%, t=-6.372, P=0.000]. The mean blood loss in the treatment group was significantly less than that of the control [(15.1?4.1) ml vs (24.7?6.6) ml; t=-7.204, P=0.000]. Conclusions Intraoperative administration of dilute pituitrin solution can reduce blood loss, operation time, and the amount of distention fluid infusion and intravasation. As a result, it is useful to avoid post-hysteroscopy complications including TURP syndrome.

Objective To evaluate the effect of sevoflurane preconditioning on intestinal injury in the patients undergoing resection for liver cancer.Methods Forty patients of both sexes,aged 20-60 yr,weighing 5075 kg,of ASA physical status Ⅱ or Ⅲ (liver function Child-Pugh grade A),undergoing resection for right liver cancer,were randomly divided into 2 groups (n =20 each):sevoflurane preconditioning group (S group) and control group (C group).Anesthesia was induced with target-controlled infusion of propofol and remifentanil.Tracheal intubation was facilitated with cisatracurium.Anesthesia was maintained with target-controlled infusion of propofol and remifentanil.S group inhaled sevoflurane with the end-tidal concentration of 2.0％ for 30 min starting from the end of intubation,followed by washout.Before induction (T0),immediately after hepatic portal was clamped (T1),at 1,3 and 6 h after occlusion of hepatic portal was released (T2-4) and at 24 h after operation (T5),arterial blood samples were obtained for determination of serum tumor necrosis factor-alpha (TNF-α),intestinal fatty acids binding protein (Ⅰ-FABP) and D-lactate levels.Results The concentrations of serum TNF-α and D-lactate started to rise at T2,peaked at T4,and started to decline at T5,and the serum Ⅰ-FABP concentrations started to rise at T2,peaked at T3,and started to decline at T4 in the two groups.The concentrations of serum TNF-α,Ⅰ-FABP and D-lactate were significantly lower in S group than in C group.Conclusion Sevoflurane preconditioning can inhibit inflammatory responses and reduce intestinal injury in the patients undergoing resection for liver cancer.

Objective To investigate the expression of microRNA-548ah (miR-548ah) and microRNA-4804 (miR-4804) in peripheral blood mononuclear cells (PBMCs) of patients with chronic hepatitis B virus infection and their clinical significance.Methods PBMCs were collected from 29 patients with chronic hepatitis B (CHB),30 hepatitis B virus carriers (HBVC),26 inactive HBsAg carriers (IASC) and 28 healthy controls in Taizhou People's Hospital during September 2012 and August 2013.Expressions of miR-548ah and miR-4804 in PBMCs were detected by fluorescence real-time quantitative RT-PCR (qRT-PCR).Receiver operating characteristic (ROC) curve was used to evaluate the expression of miR-548ah and miR-4804 in distinguishing immune tolerance phase and clearance phase of HBV infection.Pearson correlation analysis was performed to assess the correlations of miRNAs expression with clinical markers alanine aminotrans ferase (ALT) and HBV DNA loads.Results There were significant differences in expressions of miR-548ah and miR-4804 in PBMCs between CHB,HBVC,IASC groups and control group (F =28.16 and 83.17,P ＜ 0.01).Compared with control group,miR-548ah was up-regulated in CHB group(P ＜ 0.01),and down-regulated in HBVC and IASC groups (P ＜ 0.01) ; miR-4804 was up-regulated in CHB group (P ＜ 0.01),down-regulated in HBVC group (P ＜ 0.01),while there was no significant difference between IASC group and control group in miR-4804 expressions(P ＞ 0.05).The areas under ROC curve (AUCs) of miR-548ah and miR4804 in differentiation of immune tolerance and immune clearance were 0.966 and 0.997,and the sensitivities and specificities were 89.7％,96.6％ and 99.6％,100.0％,respectively.No significant correlation was found between the expression of miR-548ah,miR-4804 and ALT,HBV DNA loads (r=0.14,0.18,-0.20 and-0.19,P＞0.05).Conclusion miR-548ah and miR-4804 may be involved in the immunopathogenesis of CHB,and their expression levels in PBMCs are helpful in differentiation of immune tolerance and immune clearance in HBV infection.

Objective To develop a time saving and sensitive cell culture system based on hepatitis C virus chimera expressing enhanced green fluorescent protein(EGFP) and to facilitate the study on HCV pathogenesis and screening of anti-HCV drugs.Methods Enhanced green fluorescent protein reporter gene and a mutation V2440L that can yield higher virus titers were introduced into the C-terminus of non-structural protein 5A (NS5A) of the JFH1 viral genome by using recombinant PCR.The viral RNA was transfected into Huh7.5 cells.Viral RNA in supernatant of HCV RNA-transfected cells was determined after transfection by RT-PCR.HCV replication and infection were determined by immunofluorescence assay.IFN-α was used to evaluate the feasibility of this system for anti-HCV drugs screening.Results The viral RNA replicated efficiently in transfected cells.These cells can produce HCV-EGFP reporter virus.Viral RNA levels in supernatant were 3.06× 105 copies/ml and 7.96×106 copies/ml at 72 h and 9 d after transfection,respectively.The virus titer reached to 104 FFU/ml 9 d after transfection.The expression of EGFP was inhibited by IFN-α in a dose dependent manner in Huh7.5 cells infected by HCV-EGFP reporter virus.Conclusion The recombinant HCV JFH1-EGFP reporter gene system is a time saving,cost effective and sensitive method for studying viral replication cycles and screening of anti-HCV drugs.

ObjectiveTo develop a cell culture system with consistent expression of whole hepatitis C virus (HCV) gene and Renilla luciferase gene and to facilitate the study on HCV pathogenesis and the screening of new antiviral drugs.MethodsRenilla luciferase (RLuc) reporter gene and a mutation that could yield higher virus gene expression were introduced into the C-terminus of non-structural protein 5A (NS5A) of the JFH1 viral genome by using recombinant PCR.The viral RNA was transfected into Huh7.5 cells.Naǐve Huh7.5 cells were infected by the supernatant from the viral RNA transfected cells.HCV replication and infection were determined by virus titration,Renilla luciferase assay,immunofluorescence assay and western blotting.IFN-α was used to evaluate the feasibility of this system for anti-HCV new drug screening.ResultsThe viral RNA replicated efficiently in transfected cells.These cells could produce high titer of HCV-Rluc reporter virus and the virus titer reached to 1.5 × 104 FFU/ml at day 15 of posttransfection.The activity of Renilla luciferase was inhibited by IFN-α in a dose dependent manner in Huh7.5 cells infected by HCV-Rluc reporter virus.ConclusionThe recombinant HCV-JFH1-Rluc reporter gene system is sensitive and efficient.It can be a useful tool for high throughput screening of anti-HCV drugs.

BACKGROUND: It was reported that the reason why the fracture healing become more and more difficult with the increasing age may be associated with the varied gene regulation of bone marrow mesenchymal stem cells differentiation into osteoblast and lipoblast. However, the exact mechanism under it remains poorly understood.OBJECTIVE: To explore the association of the age-related changes on expression of peroxisome proliferator-activated receptor γ (PPARγ) and core binding α1 (Cbfα1) with the age-relatad fracture healing impairment, we studied the expression changes of PPARγ and Cbfα1 during aging fracture healing.DESIGN, TIME AND SETTING: A randomized control animal trial was performed in the Third Xiangya Hospital of Central South University between October 2007 and Febuary 2008. MATERIALS: Six male SD rats of 3 months and the other six of 23 months, randomly-selected, were divided into two groups: the old experiment group (23 months) and the young control group (3 months). METHODS: Transverse osteotomies on the middle-upper parts of left tibiae were performed to the rats, with the self-made extemal mini-fixator implanted there simultaneously. Tractions were taken twice a day (0.2 mm/d) for 14 days from day 2 till day 15 post operation. On day 15 post operation, rats were sacrificed to harvest left tibea samples. MAIN OUTCOME MEASURES: Both imaging examination and histological observation were performed to the fractures of rats. The PPARγ and Cbfα1 expression were detected with reverse transcription-polymerase chain reaction (RT-PCR) technique. RESULTS: All samples were involved in the results analyses. The imaging examination showed that many osteotylus generated between the broken ends of fractures in the young control group; that the bone formation of the young control group was much better than that of the old experiment group. The histological observation showed that large amounts of osteotylus growth occurred to every animal in the young control group and their membrane bone formation was remarkable; comparatively, the osteotylus growth in the old group was weakened dramatically, and only fibrous joints were seen. The RT-PCR detection showed that differences of significance existed in both PPARγ and Cbfα1 mRNA expressions between the two groups, with the much higher Cbfol mRNA expression in the bone marrow of rats in the young group and the much higher PPARy mRNA expression in the bone marrow of rats in the old group.CONCLUSION: Rats of different ages show different fracture healing abilities, i.e. the higher ages, the lower fracture healing abilities. In addition, expressions of PPARγ and Cbfα1 in the bone marrow of rats alter with the increasing age, which indicates that there are certain associations between the changed expression level of the two factors and the fracture healing impartment.

The Department of Surgery of PUMCH did plenty of work to optimize the teaching model,which is characterized by stage-dependent integrity and by systematic organization as a response to the challenge to students' training.

Objective To study the clinical features of complicated upper urinary tract infection in children,aiming to provide a reference for clinical diagnosis and treatment of the disease.Methods The clinical data of 68 cases with complicated upper urinary tract infection hospitalized at the Department of Urinary Surgery NO.1,Children's Hospital of Shenzhen between January 2013 and August 2015 were retrospectively analyzed.Results A total of 68 cases,in which 44 were male,24 were female,and repeated infections were found in 15 cases.Fever was the most common clinical manifestation(48 cases,70.59％),while frequent urination and odynuria were rare(9 cases,13.24％).A total of 57 strains had been cultured form the submitted specimens which were obtained from the 68 cases,including 41 strains of gram-negative bacteria,15 strains of gram-positive bacteria and 1 strain of fungus.Two different strains were cultured in 3 children.Fourteen strains of Escherichia coli had 11 extended spectrum beta lactamases (ESBLs)-positive strains.In the 11 strains of klebsiella pneumoniae,8 trains were ESBLs-positive.The drug resistant rates of gram-negative bacteria to Ampicillin and Cefuroxime sodium were both more than 90％.The sensitive rates to Piperacillin/Tazobactam was more than 90％.Thirty-one cases of complicated upper urinary tract infection were cured by administering Piperacillin/Tazobactam,while 15 cases were cured by changing Cefuroxime sodium to Piperacillin/Tazobactam according to the drug sensitivity results.Conclusions Clinical manifestations of complicated upper urinary tract infection are untypical,and fever is the most common symptom.Repeated infection is common.The gram-negative bacteria is the dominant pathogen causing the complicated upper urinary tract infection.ESBLs-positive bacteria accounts for high proportion.The drug resistance rate to penicillin and the first,second generation of the cephalosporin is high.The drug sensitive rate of piperacillin tazobactam is high,with good prognosis.

Objective To study the characteristics of peripheral blood lymphocyte subsets in patients with lung cancer,gastric cancer and breast cancer.Methods Five hundred and eight patients with cancer from the First Affiliated Hospital of Tianjin University of Traditional Chinese Medicine (256 cases with lung cancer,152 cases with gastric cancer,100 cases with breast cancer),and 50 healthy volunteers were collected.Two ml peripheral blood were obtained from these cases.The distribution of lymphocyte subsets in peripheral blood was measured by flow cytometry in these cases.The data were analyzed using SPSS 16.0 software.Results The number of lymphocytes of cancer patients was decreased,the healthy volunteers was 5125,breast cancer patient was 3642,gastric cancer patient was 3178,lung cancer patient was 2895.The case of outliers of lymphocyte subsets classified based on the three kinds of cancer was 218 (85.2％,lung cancer),133 (87.5％,gastric cancer),88 (88.0％,breast cancer).The case of outliers of T-lymphocyte subsets classified based on the three kinds of cancer was 83 (32.4％,lung cancer),32 (32.0％,breast cancer),44 (28.9％,gastric cancer).The case of outliers of CD4 +/CD8 + classified based on the three kinds of cancer was 185 (72.3％,lung cancer),108 (71.1％,gastric cancer),84 (84.0％,breast cancer).The case of outliers of natural killer-lymphocyte subsets classified based on the three kinds of cancer was 32 (12.5％,lung cancer),22 (14.5％,gastric cancer),16 (16.0％,breast cancer).The case of outliers of B-lymphocyte subsets classified based on the three kinds of cancer was 38 (14.8％,lung cancer),52 (34.2％,gastric cancer),12 (12.0％,breast cancer).Compared to healthy subjects,the CD19+％ of patients with lung cancer was decreased (12.8 ± 5.0 vs.11.5 ± 5.7,t =3.006,P =0.003);the CD4 + ％ of patients with gastric cancer was decreased (39.2 ±7.7 vs.35.3 ± 7.6,t =2.315,P =0.023);the CD19 + ％ of patients with gastric cancer was decreased (12.8 ± 5.0 vs.8.9 ± 4.2,t =3.302,P =0.010);the CD8 + ％ of patients with breast cancer was increased (24.0 ± 8.1 vs.29.1 ± 13.0,t =2.019,P =0.047).Conclusion The number of lymphocytes in cancer patients is decreased,the abnormal rates of lymphocyte subsets in the three kinds of cancer patients are higher than those in healthy volunteers,the lymphocyte subsets of different kinds of cancers perform different characteristics.

The sterile stem tips, nodular and non-nodular stem of Morinda officinalis How were cultured in vitro. The results showed that the most suitable procedure for sterilization of explants was soaking into 0.1% HgCl 2 solution for 10 to 15 minutes after pretreating with 70% ethanol for 30 seconds. MT culture medium with BA was effective to induce direct shooting of stem tips and nodular stem, the shooting rate of nodular stem being 97.5% . The optimal BA concentration was 1 mg?L -1 , the shooting rate would decrease when the concentration of BA increased. As for the induction of rooting, MT culture medium adding with 0.2 to 0.5 mg?L -1 NAA was optimal, the rooting rate being over 80.0% .