Objective To detect the sperm apoptosis rate and level of reactive oxygen species (ROS) in seminal plasma and explore their correlation among infertile males. Methods Ninety-two inferitile males were divided into varicocele (VC) group (n=32), leukocytospermia group(n=30) and the other cause group (n=30), and another 24 in vitro fertilization sperm samples were sereved as controls. The routine sperm parameters including seminal pH, sperm viability and sperm density were examined by computer assisted sperm analysis, the sperm apoptosis rate was asseseed using Annexin V/PI staining, and the ROS level in seminal plasma was detected by TBA method. The differences in seminal parameters between three infertile groups and control group were compared, and the correlation of sperm apoptosis rate with level of ROS in seminal plasma was explored in each group. Results The sperm viability of three infertile groups was significantly lower than that of control group (P<0.01). The sperm apoptosis rates and levels of ROS in seminal plasma in VC group and leukocytospermia group were significantly higher than those in control group (P < 0.05 or P < 0.01). The sperm apoptosis rate was positively correlated with the level of ROS in seminal plasma in leukocytospermia group(r=0. 573, P < 0.05). Conclusion The increased sperm apoptosis rate and level of seminal plasma ROS may be related to the infertility of patients with VC and leukocytospermia. The increased level of seminal plasma ROS may be one of the causes of increased sperm apoptosis rate in patients with leukocytospermia.

In view of the predominance problems in present experimental teaching of medical microbiology, we reformed the experimental contents and experimental teaching methods,to establish an entire experimental curriculum teaching system to be suitable for seven-year system and long educational system students,which manifests the creative teaching idea.

OBJECTIVETo evaluate the risk awareness and preparedness related to pandemic flu in China.

METHODSTwo groups of people, mainly employers and employees from enterprises, were covered in the survey, using quantitative (questionnaire) and qualitative (in-depth interview) methods. The employers and employees were from joint-ventured corporations, large state-owned corporations and private companies which were randomly selected from 7 major cities in China.

RESULTS(1) 82% of the people surveyed and interviewed had basic knowledge on pandemic flu. (2) 60% of the joint-ventured corporations had worked out or were working on their business continuity plan in the event of pandemic flu, compared to that of state-owned corporations and private companies that the figure was only 21% . (3) 67% of the joint-ventured corporations had informed their preparedness plan on pandemic flu to their employees, while that of the state-owned and private corporations, it was only 42 %. (4) About 70 % of the corporations was establishing policies for restricting travel to affected geographic areas (both domestic and international), evacuating the employees who working in or near the affected area when an outbreak began. (5) Nearly 60 % of the corporations thought annual flu vaccination was important and hence encouraging and tracking annual flu vaccination for employees. (6) 70% of the corporations paid high attention on providing sufficient and accessible supplies (e. g. hand - hygiene products, tissues and receptacles for their disposal) to control the epidemics in all business locations while nearly 76 % of the corporations were interested in purchasing commercial medical insurance.

CONCLUSIONJoint-verntured corporation were doing better than domestic corporations in terms of risk awareness and preparedness on pandemic flu, suggesting that the domestic corporation should learn from them regarding on pandemic flu preparedness to limit the negative impact of pandemic flu.

Adult ; Awareness ; Commerce ; Disease Outbreaks ; prevention & control ; Female ; Humans ; Influenza, Human ; epidemiology ; prevention & control ; psychology ; Male ; Middle Aged ; Surveys and Questionnaires ; Urban Health

The study established a UPLC-MS/MS method that is used for simultaneous determination nine major bioactive compounds of Dachengqi Tang in rat plasma. Using Aglient C18 column (2.1 mm x 50 mm,1.7 microm) was chromatographed, using methanol-5 mmol x L(-1) ammonium formate mobile phase gradient, elution 0.3 mL x min(-1). In the plasma pre-treatment process, not only the method of methanol and acetonitrile protein precipitation was investigated, and different factors extraction solvent, the type of the scroll time, the number and the type of extraction solvent, the extraction volume of the extraction solution of liquid-liquid extraction is investigated. Finally, with ibuprofen as an internal standard, using ethyl acetate liquid-liquid extraction method pretreatment blood, N2 dry reconstituted supernatant after centrifugation UPLC-MS/MS analysis, in electrospray ionization (ESI) negative mode, using multiple reaction monitoring mode for testing. The linear range of emodin, rhein, aloe-emodin, chrysophanol, magnolol, honokiol, hesperidin and hesperitin is 0.33-660, 0.40-792, 0.41-827, 0.34-680, 0.45-907, 0.46-927, 0.43-867, 0.34-683, 0.39-787 microg x L(-1) respectively, good linear relationship; and extraction recovery were greater than 69.39%, days after the day of the RSD is less than 15%. This method can be used to study the rat gastric large bearing gas after Dachengqi Tang, the simultaneous determination of nine components in plasma for its pharmacokinetics and efficacy material base to provide a theoretical basis.
Animals ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Female ; Male ; Plasma ; chemistry ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods

OBJECTIVETo explore the effects of paeoniflorin on blood brain barrier and pathological changes in brain ischemia.

METHODMice were divided into sham operation group, model group, positive control group-Deng zhanhua tablet group and three different dose (high, middle, low-dose) groups of paeoniflorin. The neurological symptoms of rats were observed. The SOD of ischemic brain tissue, MDA BBB and EAA contents were determined. The ultrastructure on the brain cells was inspected by transmission electron microscope.

RESULTPaeoniflorin had the protetive effects on 4VO model of total cerebral ischemia. Paeoniflorin could obviously increase SOD content, reduce MDA content in rat brain-tissue and alleviate oxidative stress damage by cerebral ischemia on rat brain. Paeoniflorin could improve pathological changes of cell nuclear, perikaryon, mitochondria and myelin sheath, which was the morphologic basis of the protective effect on ischemia. Paeoniflorin could alleviate the incrense of EAA content caused by and hypoxia and inhibit the excitatory neural toxicity by EAA.

CONCLUSIONPaeoniflorin has the protection effect on the brain edema after cerebral ischemia, the oxidative stress damage on brain tissue, the ultrastructure lesions of cells and the BBB. The protective mechanism may be related to inhibiting intracellular calcium overload, anti-free radical and reducing EAA content.

Animals ; Behavior, Animal ; drug effects ; Benzoates ; isolation & purification ; pharmacology ; Blood-Brain Barrier ; drug effects ; pathology ; ultrastructure ; Brain ; blood supply ; drug effects ; metabolism ; Brain Ischemia ; pathology ; physiopathology ; prevention & control ; Bridged-Ring Compounds ; isolation & purification ; pharmacology ; Glucosides ; isolation & purification ; pharmacology ; Male ; Malondialdehyde ; metabolism ; Microscopy, Electron ; Monoterpenes ; Neuroprotective Agents ; isolation & purification ; pharmacology ; Paeonia ; chemistry ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Wistar ; Superoxide Dismutase ; metabolism

OBJECTIVETo establish the HPLC fingerprints of tanshinone microemulsion.

METHODHPLC analysis was carried on Hypersil C18(4.6 mm x 250 mm, 5 microm) analytical column; the acetonitrile and 0.5% phosphoric acid solution were used as mobile phases with gradient elution at a flow rate of 1.0 mL x min(-1). The UV detection wavelength was set at 270 nm.

RESULTSix peaks on HPLC fingerprint of Tanshinone Microemulsion are indexed.

CONCLUSIONThe method developed in the present study is convenient, reliable, and can be used as a quality control method for Tanshinone Microemulsion.

Chromatography, High Pressure Liquid ; methods ; Diterpenes, Abietane ; Emulsions ; Phenanthrenes ; administration & dosage ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Quality Control ; Salvia miltiorrhiza ; chemistry

OBJECTIVETo investigate the influence of escharectomy at different time-points after burn injury on the lymphocyte apoptosis and the antigen presentation function of monocytes in peripheral blood of scalded rats.

METHODSOne hundred and thirty-six Wistar rats were randomly divided into normal control ( C,n = 8 ), scald ( S, n = 64,without treatment after scald) , A ( n = 40, with escharectomy at 36 post-burn hour( PSH) ) , B ( n = 24, with escharectomy at 72 PSH ) groups. The rats in A , B, S groups were inflicted with 30% TBSA full-thickness scald. The rats in S group were sacrificed on 6,12,24,72,120,168,216, 288 PSH, while those in A and B groups were sacrificed at 72 -288 PSH, 168 -288PSH, respectively. The rats in C group were also sacrificed as control. The apoptotic rate of peripheral lymphocytes, the positive expression rate of MHC- II in mononuclear cells, the changes in concentration of IL-4 and gamma-IFN were determined in each group. The correlation of above indices were also analyzed.

RESULTS(1) The apoptotic rate of peripheral lymphocyte in S group were increased dramatically at 6PSH, peaking at 24 PSH( 18. 19+/-1.42% ) , then decreasing gradually, reaching the lowest level at 72 PSH(8. 25+/-0.56% ) , then it increased gradually again, approaching almost the peak value at 288 PSH( 17.81 +/- 1.99% ). The values were all obviously higher than those in C group( P <0.05). The apoptotic rates of peripheral lymphocyte in A and B groups were evidently lower than that in S group ( P <0. 01). (2) The positive expression rate of MHC-II in monocyte was decreased sharply at 6 PSH, and it was 20% lower than that in C group (37. 2 +/- 2. 4% ) at 24 PSH. It then increased gradually, but it was significantly lower than that in A, B groups at 288 PSH (18. 8 +/-2. 8, P <0.01). (3) The plasma level of y-IFN in S group increased gradually from 6 PSH on, peaking at 24 PSH(440. 8 +/-25. 1 )ng/L,then decreasing gradually , and it reached the lowest level at 288 PSH (51.3 +/-37.0) ng/L. The IL-4 level in S group was increased gradually ,peaking at 288 PSH (78. 1+/-2. 8) ng/L. (4) There was negative correlation between the expression rate of MHC- II in S group and IL-4/gamma-IFN ratio in escharectomy groups during 72 - 288 PSH ( r = - 0. 96, P < 0. 05).

CONCLUSIONEacharectomy after scald can inhibit peripheral lymphocyte apoptosis, slow down the insertional tendency of IL-4/gamma-IFN , and ameliorate the antigen presentation function of monocytes. Moreover, escharectomy during shock stage can markedly promote the immune function of monocytes.

Animals ; Antigen Presentation ; Apoptosis ; Burns ; immunology ; pathology ; surgery ; Genes, MHC Class II ; Interferon-gamma ; blood ; Interleukin-4 ; blood ; Lymphocytes ; cytology ; immunology ; Male ; Monocytes ; immunology ; Rats ; Rats, Wistar ; Shock, Traumatic ; immunology ; pathology

OBJECTIVETo assess the diagnostic value of multiplex ligation-dependent probe amplification (MLPA) for detection of common chromosome aneuploidy in amniotic fluid (AF) cells in order to obtain an accurate, rapid, cost-effective and high-throughput method in routine prenatal clinical practice.

METHODSThe MLPA test was performed on 500 AF samples by using kit P095 and the results were obtained by using analysis software RH-MLPA-v511. The results were compared with that from fluorescence in situ hybridization (FISH) and traditional karyotyping (TK). The technical critical issues were analyzed in routine diagnostic application.

RESULTSThe absolute specificity and sensitivity of the MLPA test to detect the aneuploidy were 100%. For the 500 AF samples, the success rate of the MLPA tests was 97%. Among them 92% were finished within three working days and 5% required more days for repeating. The test failure rate was 3%. The results confirmed that for the 38 detectable aneuploid samples, the probe reliability weighted mean ratio values were more than 4SD compared to normal diploids and the 2 suspected trisomy samples were more than 2SD. In this study, authors analyzed hybridization efficiencies of 8 probes for chromosome 21, and the presence of a trisomy was considered if at least 4 of the 8 probes gave probe ratio of >1.3.

CONCLUSIONThedata suggested that MLPA is a rapid, simple and reliable method for large scale testing for aneuploidy of chromosomes 13, 18, 21, X, or Y in AF. The MLPA technology is complementary to AF culture and valuable for prenatal diagnosis.

Amniotic Fluid ; cytology ; Aneuploidy ; Chromosomes, Human, Pair 21 ; Female ; Humans ; Nucleic Acid Amplification Techniques ; methods ; Pregnancy ; Prenatal Diagnosis ; methods ; Sensitivity and Specificity ; Trisomy ; diagnosis ; genetics

The formalin test is a commonly used animal model of acute and tonic pain. However, the molecular targets of formaldehyde (FA, the main ingredient of the formalin solution) on primary nociceptor cells remain controversial. In this report, the effects of FA on electrophysiologically-identified primary nociceptor cells were evaluated in vitro and the roles of the vanilloid receptor TRPV1 in FA-produced activation of primary nociceptors were also examined at both cellular and behavioral levels. Of 92 acutely dissociated dorsal root ganglion (DRG) cells recorded by current patch-clamp technique, 34% were discharged by FA application with the mean onset latencies of the first action potential (AP) being (367.34+/-32.96) s. All the FA-sensitive cells were identified as nociceptor cells by their distinguishable features of AP including longer duration, existence of a hump (a shoulder or inflection) on the repolarizing phase, and longer after-hyperpolarization of APs. Co-application of capsazepine (CPZ), a competitive antagonist of TRPV1 receptors, could block FA-evoked firing with partial inhibition on the membrane depolarization of all cells tested. Of another 160 cells examined by confocal calcium imaging, 32% were shown to respond to FA with an intracellular Ca(2+) rise. Of 51 FA-sensitive cells, 67% were suppressed by CPZ, suggesting partial involvement of TRPV1 in mediation of the FA-evoked intracellular Ca(2+) rise. Under voltage-clamp mode, 41% of DRG cells were evoked to give rise to inward current with the remaining 59% being unchanged. In separate experiments on the other 56 FA-sensitive cells, concentration-dependent increase in the FA-evoked current amplitude was demonstrated. In comparison with controls, the FA-evoked inward current could be significantly suppressed by CPZ that was further enhanced by HC-030031, a TRPA1 selective antagonist. Finally, local effects of CPZ were confirmed in the formalin test and it was shown that the formalin-induced paw flinches were strongly suppressed by CPZ in phase 1 but with phase 2 being significantly suppressed only during 25-55 min. It is therefore concluded that FA can directly activate a subpopulation of primary nociceptor cells and the FA-induced AP discharges are likely to contribute mainly to phase 1, but not phase 2 of the formalin-induced nociception. The activation of primary nociceptor cells by FA is likely to be mediated, at least in part, through TRPV1 and/or TRPA1 receptors.
Acetanilides ; pharmacology ; Action Potentials ; Animals ; Capsaicin ; analogs & derivatives ; pharmacology ; Formaldehyde ; pharmacology ; Ganglia, Spinal ; physiology ; Nociceptors ; physiology ; Pain ; physiopathology ; Pain Measurement ; Patch-Clamp Techniques ; Purines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; TRPV Cation Channels ; physiology

Purpose: The objective of the current study was to examine the potential effects of surgery start times (morning vs. afternoon) on the long-term prognosis of patients after hepatic resection (HR) for hepatocellular carcinoma (HCC). Methods: All eligible patients were divided into 2 groups according to the start time of surgery: group M (morning surgery, 8 AM–1 PM) and group A (afternoon surgery, 1 PM–6 PM). Clinicopathologic and surgical parameters, as well as oncologic outcomes were compared between the 2 groups. Results: In total, 231 patients were included in the study. There was no difference in age, body mass index, comorbidities, tumor size, tumor location, tumor stages, surgical procedures, or surgical margin between morning and afternoon surgery (all P > 0.05). In contrast, patients in group M experienced longer operation duration than those in group A (median, 240 minutes vs. 195 minutes, P = 0.004). However, no differences of overall survival were observed between morning and afternoon surgery groups in the whole cohort or stratified by surgical margin (all P > 0.05). Conclusion Surgery start times during the work day have no measurable influence on patient outcome following curative HR for HCC, indicating good self-regulation and professional judgment of surgeons for progressive fatigue during surgery.