In this study, a fast carbonyl reductases colorimetric screening method for discovering stereoselective carbonyl reductases was established by combining the reverse alcohol oxidation with the azoreductase-catalyzed reduction of azo dye. When azo dye (Orange I , 4-(4-hydroxy-1-naphthylazo) benzenesulfonic acid) and azoreductase (AzoB) were added into the reaction system of alcohol oxidation catalyzed by carbonyl reductase, the produced NAD(P)H served as electron donor for the azoreductase to reduce the azo dye, resulting the color fade. Hence, the carbonyl reductases can be screened by the obvious color change. When chiral alcohol was used as the substrate, the activity and stereoselectivity of carbonyl reductases can be screened at the same time.
Alcohol Oxidoreductases ; chemistry ; Alcohols ; chemistry ; Azo Compounds ; chemistry ; Coloring Agents ; chemistry ; High-Throughput Screening Assays ; NADH, NADPH Oxidoreductases ; chemistry ; NADP ; chemistry ; Oxidation-Reduction ; Stereoisomerism

Objective To investigate the traffic capacity of ambulances in Beijing,and to explore factors and methods to resolve this problem.Method A survey on all the ambulances on-duty in Beijing from 17th August to 17th September 2006 was conducted by questionnaires.Results The average speed of ambulances in Beijing was 32.07 km/h.There were statistically significant differences in terms of the areas and time, respectively.Conclusions The traffic jam of Beijing was serious.The traffic capacity of ambulances was far from being ideal,which restricted the operation of 120 ambulances.It is difficult to satisfy the needs of Beijing 2008 Olympic games and it is time for the relevant authorities to search for appropriate methods and solve this problem.

OBJECTIVETo evaluate the clinical effect of arthroscopic excision of the os subfibulare in anterior-lateral ankle pain.

METHODSFrom December 2005 to Augest 2014, 16 patients suffering from pain associated with an os subfibulare in the anterior-lateral side of their ankles were reviewed. Among the patients,11 patients were male and 5 were female, with a mean age of (33.5 ± 15.6) years old. The mean maximum diameter of os subfibulare was (0.70 ± 0.26) cm. All the patients underwent excision of the osseous fragments, and had anatomic reconstruction of the anterior talofibular ligament if the anterior-lateral ankle was instable. The average follow-up period was (18.0 ± 4.5) months. To analyze the surgical outcome, American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot pain and function scales,visual analogue scale (VAS) and Tegner activity scale were assessed preoperatively and postoperatively.

RESULTSAOFAS scales were preoperative 60.15 ± 14.52 and postoperative 92.35 ± 5.73. There was a significant difference between them (t = -8.251, P = 0.000). The mean VAS score were preoperative 7.35 ± 0.46 and postoperative 2.45 ± 0.98. Statistical significance was also notable (t = 18.105, P = 0.000). Tegner score was significantly increased from preoperative 2.87 ± 1.12 to postoperative 5.78 ± 1.06 (t= -7.548, P = 0.000).

CONCLUSIONIrrespective of the size of os subfibulare, in patients with pain or instability associated with the os subfibulare, arthroscopic excision combined with reconstruction of ther anterior talofibular ligament or not was effective in restoring ankle function and eliminating pain.

Adult ; Ankle Injuries ; surgery ; Ankle Joint ; surgery ; Arthroscopy ; methods ; Female ; Fibula ; surgery ; Humans ; Lateral Ligament, Ankle ; surgery ; Male ; Middle Aged

OBJECTIVETo explore the inhibition of Hedyotis diffusa Willd Injection (HDI) on the proliferation of RPMI 8226 cells and its mechanisms.

METHODSThe inhibition of HDI on the proliferation of RPMI 8226 cells was detected by MTT and the drug concentrations for further researches were screened out. The apoptosis rate was detected using Annexin V-PI of flow cytometry. The cell cycle distribution was detected by PI. The expressions of adhesion molecule FITC-CD44 and PE-CD49d were detected. The IL-6 and VEGF concentrations of cell supernatants were tested by ELISA. The mRNA expressions of Bax, Bcl-2, Caspase-3, Survivin, IL-6, and VEGF were detected by RT-PCR.

RESULTSHDI could inhibit the proliferation of RPMI 8226 cells. Meanwhile, it induced their early apoptosis, arresting them at G1 phase in a concentration-dependent manner. The VEGF concentrations were down-regulated after acted by 0, 20, 40, and 60 microL/mL HDI in a dose-dependent manner (P< 0.01). The IL-6 content increased (P<0.01). The expressions of CD44 and CD49d were up-regulated in a concentration-dependent manner. After acted by 40 microL/mL HDI, the Survivin mRNA level was significantly downregulated (P<0.01), the mRNA levels of Bcl-2, IL-6, and VEGF were significantly up-regulated (P<0.01), but the up-regulation of Bax and Caspase-3 mRNA levels were not so obvious (P>0.05).

CONCLUSIONSHDI could inhibit the proliferation of RPMI 8226 cells. Its mechanisms might be correlated with early apoptosis induction, G1 phase arresting, VEGF secretion lowering, and Survivin mRNA transcription level down-regulating.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Hedyotis ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Interleukin-6 ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism

The uracil in DNA comes from either the misincorporation of dUTP in place of dTTP or deamination of cytosine. In the latter case, it can result in a GC to AT transition mutation if the uracil is not removed before DNA replication. Base excision repair (BER) is a major pathway for removing DNA lesions arising from endogenous processes as well as those induced by exposure to exogenous chemicals or irradiation. BER is initiated by DNA glycosylases that excise aberrant bases from DNA by cleavage of the N-glycosidic bond linking to the base of its deoxyribose sugar. Uracil N-glycosylase (UNG) is the enzyme responsible for the first step in the BER pathway that specifically removes uracil from DNA. The UNG gene undergoes both temporal and spatial regulation mainly at the level of transcription. Normally cancer cells undergo over-proliferation and up-regulate their UNG during tumorigenesis. In this study we examine the correlation between UNG level and carcinogenesis, and explore the possibility of using UNG as a marker for cancer diagnosis. Human UNG gene was amplified from the total RNA of the human choriocarcinoma cell line, JEG-3, by RT-PCR. After purification, the 942bp full-length UNG cDNA coding sequence was digested with EcoR I and Sal I, and cloned into the digested pET-21 to construct a recombinant vector, pUNG. The UNG protein was expressed under the control of T7 promoter in E. coli BL21 (DE3) cells induced with IPTG. After ultrasonic treatment, the cell lysate and precipitate were analyzed by SDS-PAGE and a 39kD band was detected. The plasmid was serially diluted at appropriate concentrations and employed as standards in the subsequent quantification. Total RNAs were extracted from 18 pairs of clinical samples, each pair contains a sample of esophageal squamous cell carcinoma (ESCC) tissue and its surrounding normal esophageal epithelia. The copy numbers of UNG mRNA in these RNA samples were determined by real-time quantitative RT-PCR using a Lightcycler (Roche). UNG was present in 13 cases of ESCC (13/18, n = 18) but absent in all of the normal tissues. The results indicated that there was a correlation between high level of UNG expression and the carcinogenesis of ESCC.
Carcinoma, Squamous Cell ; genetics ; metabolism ; Cell Line, Tumor ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Esophageal Neoplasms ; genetics ; metabolism ; Humans ; In Vitro Techniques ; Reverse Transcriptase Polymerase Chain Reaction ; Uracil-DNA Glycosidase ; genetics ; metabolism

OBJECTIVETo evaluate the effects of the Chinese herbal formula Wuzi Yanzong Pill (, WYP) on the spermatogenesis and specific secretory functions of Sertoli cells in rat model and to investigate the underlying mechanism.

METHODSFive groups of male Sprague-Dawley rats including the control group, the model group, the low-dose WYP group, the medium-dose WYP group and the high-dose WYP group (5 in each group) were treated daily with vehicle, multiglycosides of Tripterygium wilfordii Hook f (GTW) either alone (20 mg/kg) or followed by WYP (0.5, 1.0, or 2.0 g/kg daily), respectively for 30 days. Serum levels of follicle-stimulating hormone (FSH), inhibin B (INHB) and testosterone (T) were evaluated using enzyme-linked immunosorbent assay. Androgen-binding protein (ABP) gene expression and transferrin (TF) protein expression in testis tissue specimens of all rats were determined using real-time reverse transcriptase polymerase chain reaction and Western blotting analysis, respectively. Histopathological alterations in the testis were determined using Johnsen's score.

RESULTSThe toxicity of GTW towards Sertoli cell secretory functions and spermatogenesis was accompanied by increased serum FSH concentrations and decreased INHB and T concentrations. Upregulated ABP mRNA levels, and decreased TF protein expression and Johnsen's scores were detected in the model group compared with the control group P<0.05 or P<0.01). Oral high-dose WYP administrations to GTW-treated rats effectively alleviated all of the GTW-induced changes in specific secretory functions of Sertoli cells (ABP, INHB and TF). Furthermore, serum T level and Johnsen's score of the testis increased greatly compared with the model group (P<0.01).

CONCLUSIONWYP has the ability to improve the spermatogenesis, possibly through modulating the secretory proteins expression of Sertoli cells.

Androgen-Binding Protein ; genetics ; metabolism ; Animals ; Blotting, Western ; Drugs, Chinese Herbal ; pharmacology ; Follicle Stimulating Hormone ; blood ; Gene Expression Regulation ; drug effects ; Inhibins ; blood ; Male ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sertoli Cells ; drug effects ; secretion ; Spermatogenesis ; drug effects ; Tablets ; Testis ; cytology ; metabolism ; Testosterone ; blood ; Transferrin ; metabolism

OBJECTIVETo investigate clinical effects of auricular acupoint (AA) in the treatment of analgesia during perioperative period in total hip arthroplasty.

METHODSFrom March 2008 to August 2010, 60 patients with late osteonecrosis of the femoral head were treated by total hip arthroplasty and randomly divided into auricular acupuncture (AA) group and control group, 30 patients in each group. There were 11 males and 19 females in the AA group,with an average age of (60.93 +/- 5.90) years; the patients were treated with auricular acupuncture on the point of Shenmen, Subcortex, Kidney and hip joint for four times a week. There were 12 males and 18 females in control group, with an average age of (59.87 +/- 6.21) years; while the patients without auricular acupuncture. VAS score was used to evaluat the degree of pain; Harris score was used to evaluat the function of hip joint. All patients received patient controlled analgesia pump (PCA) for 48 hours after surgery (400 ml liquids were in PCA pump, including 800 mg tramadol and 0.8 mg fentanyl). The dosage of liquids and adverse reaction of PCA pump in different time were recorded.

RESULTSThe VAS score on the 3rd, 4th, 5th and 7th day separately was (3.61 +/- 0.29), (3.59 +/- 0.30), (2.97 +/- 0.26), (2.29 +/- 0.45), and lower than control group, which separately was (4.19 +/- 0.28), (4.00 +/- 0.31), (3.15 +/- 0.29), (2.83 +/- 0.31). The dosage of PCA in AA group separately was (72.27 +/- 8.06), (60.40 +/- 8.16), (44.13 +/- 4.75), (40.40 +/- 3.69), and less than control group, which was (86.27 +/- 8.51), (73.87 +/- 8.32), (54.53 +/- 5.20), (44.67 +/- 6.31) on the time of 0-12, 12-24, 24-36 h and 36-48 h after surgery. During the using of PCA, nausea and vomiting occurred in 5 cases, less than control group (21 cases). Harris score in AA group (78.90 +/- 5.14) was higher than control group (73.37 +/- 5.99) 2 weeks after operation.

CONCLUSIONAuricular acupuncture can reduce postoperative pain, reduce the usage of analgesic and complications, such as nausea and vomiting, improve the function of hip joint after operation.

Acupuncture Analgesia ; Acupuncture, Ear ; Adult ; Aged ; Analgesia, Patient-Controlled ; Arthroplasty, Replacement, Hip ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Pain Management ; Pain, Postoperative ; therapy ; Perioperative Period

OBJECTIVETo probe into the diagnosis and treatment of urachal remnants with heterotopic sinus.

METHODSTwo cases with penis dorsal drainage for 5 approximately 6 years were diagnosed as urachal anomaly by sinus probing, methylene blue injection test, cystoscope examination, and type B ultrasonic examination, and both received surgical removal of the urachus and all the abnormal tissues associated with it.

RESULTSSurgery and pathology proved urachal cyst with communication drainage in both of the patients. Follow-up survey showed no recurrence and malignancy.

CONCLUSIONSinus probing, mythylene blue injection test and type B ultrasonic examination are effective methods for the diagnosis of urachal remnants with heterotopic sinus. Surgical treatment should include radical removal of the urachus and the abnormal tissues associated with it.

Adult ; Cystoscopy ; Humans ; Male ; Methylene Blue ; Urachal Cyst ; diagnosis ; pathology ; Urachus ; abnormalities ; pathology ; surgery

OBJECTIVETo compare the outcomes of intracytoplasmic sperm injection (ICSI) with retrieved epididymal and testicular sperm for obstructive azoospermia and with ejaculated sperm for severe oligozoospermia and asthenospermia.

METHODSWe retrospectively analyzed 431 ICSI cycles, which were divided according to sperm sources into Groups A (n=287 in patients with severe oligozoospermia or asthenospermia using ejaculated sperm), B (n=109 in obstructive azoospermia patients with sperm retrieved by percutaneous epididymal sperm aspiration, PESA) and C (n=35 in obstructive azoospermia patients with sperm retrieved by testicular sperm extraction, TESE). Comparisons were made among the three groups in the rates of embryo implantation, fertilization, pregnancy, cleavage, and miscarriage.

RESULTSGroup A showed statistically significant differences from Groups B and C in the rates of embryo implantation and pregnancy (18.46% vs. 25.23% and 28.76%, 31.23% vs. 42.16% and 39.39%, P < 0.05). But no significant differences were seen in the rates of fertilization, cleavage and miscarriage among the three groups (P > 0.05).

CONCLUSIONThe rates of embryo implantation and clinical pregnancy are higher in patients with obstructive azoospermia than in those with severe oligozoospermia or asthenospermia after ICSI with ejaculated sperm.

Azoospermia ; therapy ; Epididymis ; cytology ; physiopathology ; Female ; Humans ; Male ; Oligospermia ; therapy ; Pregnancy ; Pregnancy Outcome ; Retrospective Studies ; Sperm Injections, Intracytoplasmic ; methods ; Spermatozoa ; Testis ; cytology ; physiopathology

Objective To understand the schistosomiasis control knowledge,attitude,and practice(KAP),and influenc-ing factors of behaviors among residents in Jiangsu Province,so as to provide the evidence for making effective health education and health promotion models. Methods The probability proportionate to size sampling(PPS)and multi-stage sampling meth-ods were adopted to sample the research objects. A questionnaire survey of schistosomiasis control KAP was conducted in the res-idents of 16 to 69 years old in schistosomiasis endemic areas of Jiangsu Province,and the results were statistically analyzed. Re-sults The total awareness rate of the participants was 95.98%for schistosomiasis control knowledge. The correct rates of atti-tude and practice were 89.06%and 77.43%,respectively. The awareness/correct rates of knowledge,attitude and practice re-duced in turns significantly(χ2=1282.96,P<0.01). The knowledge awareness rate of fishermen and boatmen was 90.98%, but their attitude correct rate was only 53.81%(χ2=120.52,P<0.01). The unconditional logistic regression analysis showed that with the education level increasing,their practice correct rate rose,and the participants with the college degree or above had a higher correct rate compared to illeterate ones(OR=6.411,95%CI:4.896-8.395). The practice correct rate of the fisher-men and boatmen was only 5.1%of the rate of the farmers(OR=0.051,95%CI:0.029-0.091). Conclusions The total aware-ness rate of basic knowledge of schistosomiasis prevention and control in the residents of Jiangsu Province has reached the re-quirements in the"National Schistosomiasis Control Long-term Planning Outline(2004-2015)",but the correct rate of behav-iors is low. The education level,occupation and residential areas affect the health behaviors of schistosomiasis prevention and control. Therefore,it is necessary to carry out targeted health promotion activities to promote the formation of healthy lifestyle and behaviors.