Objective To analyze the epidemiological features and influencing factors of human brucellosis in Qinghai province,and to provide scientific basis for prevention and control of brucellosis.Methods From 2006 to 2010,select the high incidence areas of brucellosis in Qinghai province and five counties(Henan,Dari,Tianjun,Ping'an and Haiyan counties) included in the “Central Subsidies to Local Public Health Special Fund Human Brucellosis Prevention and Control Projects” for the survey point,as well as high-risk employees from Qinghai Biological Pharmaceutical Factory were investigated.Combined with epidemiological questionnaire investigation [done according to the “National Human Brucellosis Surveillance Program(Trial)”],clinical symptoms and signs,confirmed human brucellosis patient were tested by intradermal allergy test,rose bengal plate agglutination test(RBPT) and standard tube agglutination test (SAT),in accordance with “Diagnostic Criteria and Principles of Management of Brucellosis” (GB 15988-1995) and“Diagnostic Criteria for Brucellosis” (WS 269-2007).Results Of 8368 serum samples detected,347 were RBPT positive,and the positive rate was 4.15％;5346 serum samples were tested by SAT,180 were positive,and the positive rate was 3.37％.In June 2009,112 employees in Qinghai Biological Pharmaceutical Factory were investigated on a follow-up survey,83 were RBPT positive,the positive rate was 74.11％; 58 were SAT positive,the positive rate was 51.79％.Eight of them were new cases and 4 were chronic brucellosis.Twenty five new cases were reported between 2006 and 2010.The peak incidence was from March to July.Most of the cases were herdsmen.ConclusionStrengthening animal quarantine,strengthening public education,and improving protection awareness,can effectively control the disease brucellosis.

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.

Objective To investigate the effect of local injection of Botulinum toxin type A(BTXA) on spasticity and function of the affected upper limb in stroke patients.Methods A total of 32 stroke patients were re- cruited and randomly divided into two groups:a BTXA group and a control group.All the patients had spasticity of upper limb muscles,which scored grade 2 to 3 with the Modified Ashworth Scale(MAS) ,and decreased elbow joint range of motion.The 16 patients in the BTXA group received BTXA injection in the biceps brachii muscles and flexor muscles of forearm on 10～15 points,while those in the control group did not.All the patients in both groups were treated with rehabilitation training techniques.The MAS,Fugl-Meyer upper limb function assessment and Barthel In- dex were employed to evaluate the changes of muscle tone,upper limb function and activity of living (ADL)perform- ance of the patients before injection and at 1st,2nd,6th 12th weeks after injection.Results The therapeutic effect between the BTXA group anti control group was significantly different in terms of biceps muscle tone,the scores of Fugl-Meyer upper limb function assessment and Barthel Index.Compared with preinjection,muscle tone was de- creased significantly and ADL performance was improved after injection in BTXA group.The effects of BTXA lasted more than 12 weeks.Conclusion Intramuscular muhipoint injection of BTXA was useful in reducing muscle spas- ticity,and was helpful for increasing motor ability of the affected upper limb and ADL performance of the stroke pa- tients.

OBJECTIVETo investigate the presence of pathogenicity island (PAI)-associated genes in the enterococcal isolates.

METHODSUsing PCR and hybridization methods, PAI-associated genes were detected in 155 enteococcal strains isolated from clinical patients and healthy individuals.

RESULTSAmong the 155 enterococcal isolates, 137 (88.39%) carried at least one of PAI-associated genes, namely hyd (positivity rate of 81.94%), psaA (78.06%), nuc (57.42%), esp (53.55%), cylB (52.90%), and gls24-like (38.06%) genes. Expect for esp gene, the other 5 genes showed higher positivity rates in the E. faecalis strains than in the E. faecium strains, and this difference was statistically significant for the genes nuc, cylB, and gls24-like. The positivity rates and the number of these genes in the E. faecalis from clinical isolates were both significantly higher than those in the strains isolated from healthy individuals.

CONCLUSIONThe data show a wide distribution of the PAI-associated genes among the enterococcal strains, and E. faecalis strains are more likely than E. faecium strains to be positive for the 6 genes, which are present at significant higher rates in the clinically isolated samples than in that from healthy individuals.

Bacterial Proteins ; chemistry ; genetics ; Enterococcus ; genetics ; isolation & purification ; pathogenicity ; Enterococcus faecalis ; genetics ; isolation & purification ; pathogenicity ; Genomic Islands ; genetics ; Gram-Positive Bacterial Infections ; microbiology ; Humans ; Membrane Proteins ; genetics ; Virulence ; genetics

The only difference of primary structure between single-chain prourokinase (pro-UK or scu-PA) and two-chain urokinase (UK or tcu-PA) is the cleavage of a single peptide bond (Lys158-Ile159) and transform scu-PA into its active two-chain form. A 13-peptide (Thr-Leu-Arg-Pro-Arg-Phe-Lys-Ile-Ile-Gly-Gly-Glu-Cys), which spans the cleavage peptide bond, was synthesized and linked to KLH (Keyhole limpet hemocyanin). The Balb/c mice were immunized by the conjugated protein with proper adjuvant. According to the Kohler and Milstein's methods, a hybridoma cell line G7 secreting monoclonal antibody specific for scu-PA was obtained. The anti-scu-PA McAb, purified from the supernatant of porous microcarrier hybridoma cell culture, was conjugated to CNBr-activated Sepharose 4B to prepare an immuno-affinity chromatography column. The u-PA was purified only by this affinity column from the supernatant of cultivating the u-PA-producing recombinant CHO cell, the u-PA recovery ratio is 90.4%, the purification factor was about 50, with the specific activity of 1.2 x 10(5) IU/mg, the scu-PA ratio in the u-PA product was 96.3%. Compared to immuno-affinity chromatography, the 3-step process for purifying u-PA (cation-exchange column, gel filtration column and benzamidine affinity column) has a u-PA recovery ratio of about 65%, with a specific activity of 1.0 x 10(5) IU/mg, and an scu-PA ratio of about 90%. These results showed that immuno-affinity chromatography is simple to recover u-PA and effective to separate scu-PA from tcu-PA.
Animals ; Antibodies, Monoclonal ; immunology ; isolation & purification ; Chromatography, Affinity ; Enzyme-Linked Immunosorbent Assay ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; immunology ; isolation & purification ; Urokinase-Type Plasminogen Activator ; immunology ; isolation & purification

Objective To explore the importance and methods of retaining articularis during pri- mary total hip arthroplasty(THA)and reconstruct soft tissue balance of hip joint after THA.Methods From February 2003 to August 2005,41 eases(43 hips)including 19 males and 22 females at age of 46- 80 years(mean 66.5 years)were treated with THA with retained capsule(Group R)and other 42 cases (44 hips)including 20 males and 22 females at age of 43-80 years(mean 64.3 years)with standard THA (Group S).Preoperative diagnosis found femoral neck fractures(GardenⅢⅣ)in 13 cases(13 hips)in Group R and 14(14 hips)in Group S;acetabular dysplasia(CroweⅢ)in 9(9 hips)in Group R and 8 (hips)in Group S;Osteoarthritis in 6(8 hips)in Group R and 7(8 hips)in Group S;and femoral head osteonecrosis(FicatⅢⅣ)in 13(13 hips)in Group R and 13(14 hips)in Group S.There were 13 hips of cement prostheses in Group R and 11 in Group S,8 cementless prostheses in Group R and 8 in Group S, 22 cement and cementless prostheses in Group R and 23 in Group S.Gibson's approach was used in both groups.Group R used the method of retaining capsule and little supination muscles during the operation to reconstruct responsibly soft tissue balancing of postoperation for THA.For comparison,Group S used the method of standard which resected a lots of capsule and didn't reconstruct it.The comparative items between Group R and Group S included incisional length,operative time,operative bleeding,drainage transfusion, infection,dislocation,postoperation standing,postoperation walking and Harris's score.Results All cases in Group R and Group S were followed for 6-22 months(mean 16.5 months in Group R and 16.7 months in Group S).There was significantly statistical difference upon interoperative and postoperative data between Group R and Group S.The result of Group R was significantly better than that of GS.Conclu- sion Retaining articularis during primary THA can minimize operative trauma,reconstruct soft tissue bal- ance and augment hip stability to get postoperative functional recovery.

OBJECTIVETo study the immunoregulatory effect of hepatitis B virus (HBV) e antigen (HBeAg) on peripheral blood monocytes (PBMCs).

METHODSPBMCs were isolated from patients with chronic hepatitis B (CHB; both HBeAg- and HBeAg+) and healthy controls, and cultured with recombinant HBeAg. The HBeAg-induced changes in expression of PD-1/PD-L1 were measured by flow cytometry of the cells and in secreted cytokines were measured by enzyme-linked immunosorbent assay of the supernatants. Comparisons between two groups were made by the independent-samples t-test; the relationship between PD-1/B7-H1 level and HBV DNA copy number was evaluated by Spearman's correlation analysis.

RESULTSExposure to HBeAg led to a significant decrease in CD3+CD4+ T lymphocyte-specific expression of IFNa for both the CHB patients' and healthy controls' samples (t = 2.382 and -4.190 respectively, P less than 0.01). For the HBeAg- CHB patients' and healthy controls' samples, the HBeAg exposure led to increased levels of secreted cytokines IL-6, IL-10 and TNFa (t = 2.504, 3.583 and 4.324, P less than 0.01 and t = 3.542, 6.246 and 5.273, P less than 0.01 respectively) and of CD14+ PBMC-specific expression of PD-L1 (t = 4.815 and 3.454, P less than 0.05 respectively). Compared to the HBeAg-negative CHB patients' and healthy controls' samples, the HBeAg+ CHB patients' samples had significantly lower CD3+CD4+ T cell-specific expression of IFNa (t = -3.177 and -4.541, P less than 0.01 respectively), but significantly higher levels of secreted IL-4 (t = 3.382 and 4.393, P less than 0.01 respectively), of CD3+ T cells-specific expression of PD-1/PD-L1 (t = 4.755, 2.942 and 4.518, 4.595, P less than 0.01 respectively), and of CD14+ T cells-specific expression of PD-L1 (t = 5.092 and 5.473, P less than 0.01 respectively). The CD3+ T cells-specific expression of PD-L1 was significantly higher in the samples from HBeAg- CHB patients than from the healthy controls (t = 3.214, P less than 0.01).

CONCLUSIONHBeAg was able to down-regulate the production of Th1-type cytokines (IFNgamma), and up-regulate the secretion of Th2-type cytokines (IL-6, IL-10) and the expression of PD-1/PD-L1on monocytes. These changes are conducive to the formation of immune tolerance to HBV. Therefore, HBeAg may play an important role in immune tolerance to chronic HBV infection.

Adult ; Case-Control Studies ; Cells, Cultured ; Female ; Hepatitis B e Antigens ; genetics ; immunology ; Hepatitis B, Chronic ; blood ; immunology ; Humans ; Interferon-gamma ; immunology ; Interleukin-10 ; immunology ; Interleukin-6 ; immunology ; Leukocytes, Mononuclear ; immunology ; metabolism ; Male ; Middle Aged ; Recombinant Proteins ; immunology ; Th1 Cells ; immunology ; Th1-Th2 Balance ; Th2 Cells ; immunology

OBJECTIVETo investigate the antimicrobial resistance of clinical isolates of Stenotrophomonas matophilia (SMA) and the mechanisms of their drug resistance.

METHODSDisc diffusion method (NCCLS) was used to detect the resistant patterns of 88 initial SMA isolates resistant to 12 antibiotics isolated from a local hospital in the past 4 years. PCR was used to detect the 7 aminoglycosides modifying enzymes genes (AME) against amikacin and gentamicin. Metal-beta-lactamases (MBLs) were screened by synergic method, and extended-spectrum beta-lactamases (ESBLs) were detected by double-disk synergy test.

RESULTSThe resistance rates of the SMA isolates were 0%-9.7% to minocycline, 12.5%-22.6% to ticarcillin-clavulanic acid, 12.5%-28.6% to levofloxacin, 18.8%-33.3% to doxycycline, 18.8%-40% to sulfamethoxazole compound, 50%-65.7% to ciprofloxacin, 50%-66.7% to cehazindme, 54.8%-66.7% to amikacin, 75%-100% to gentamicin, 81.3%-100% to piperacillin, 87.5%-100% to aztreonam and 93.5%-100% to imipenem. Aac(3)-I and ant(4')-II were not detected in these strains. The positive rates of the other 5 AME genes of aac(3)-II, ant(2'')-I, aac(6')-I, aac(3)-III, aac(3)-IV were 2.3%, 5.7%, 8%, 10%, and 10%, respectively. SMA strains producing ESBLs were found at the rate of 38.6%; 25% of the strains were MBL-producing, and 13.6% produced both ESBLs and MBLs.

CONCLUSIONMost of the SMAs we isolated are multidrug-resistant through various mechanisms. The choice of antibiotics should be made according to the susceptibility results.

Amikacin ; pharmacology ; Drug Resistance, Multiple, Bacterial ; Gentamicins ; pharmacology ; Humans ; Imipenem ; pharmacology ; Microbial Sensitivity Tests ; Stenotrophomonas maltophilia ; drug effects ; isolation & purification

OBJECTIVETo evaluate the protective efficacy of plague subunit vaccine, BALB/c mice, guinea pigs and rabbits were used in this study.

METHODSGroups of mice (10 per group), guinea pigs (14 per group) and rabbits (6 per group) were immunized with F1 + rV270 vaccine, EV76 vaccine and alum adjuvant by intramuscular route, respectively. Serum antibody titres of mice, guinea pigs and rabbits were determined by ELISA and the immunized animals were challenged with 10(6) CFU of Y. pestis strain 141 at the 8th week after the primary immunization.

RESULTSThe immunized mice, guinea pigs or rabbits with subunit vaccine developed anti-F1 IgG titre of 41 587.3 +/- 2.1, 11 543.7 +/- 2.1 or 522.4 +/- 22.4 and elicited statistical anti-F1 IgG titre difference among them (F = 17.58, P < 0.01). The immunized mice, guinea pigs or rabbits with subunit vaccine had anti-rV270 IgG titre of 15 748.7 +/- 1.6, 12.6 +/- 1.4 or 1648.0 +/- 5.0 and induced statistical anti-rV270 IgG titre difference among them (F value was 16.34, P < 0.01). There was significant anti-F1 IgG titre difference among mice, guinea pigs and rabbits immunized with EV76 vaccine that developed anti-F1 IgG titre of 913.4 +/- 4.5, 937.0 +/- 2.0 or 342.0 +/- 12.0 (F = 23.67, P < 0.01), whereas the immunized mice, guinea pigs and rabbits with EV76 vaccine developed anti-rV270 IgG titre of 12.0 +/- 1.0, 447.0 +/- 10.0, 40.0 +/- 11.0 and there was no anti-rV270 IgG titre difference between them (F = 2.20, P = 0.1314). The immunized mice with subunit vaccine developed significantly higher anti-F1 IgG titres than immunized guinea pigs and rabbits (q value was 30.57 and 19.04, respectively, P < 0.01), and there were no anti-F1 IgG titre differences between the immunized guinea pigs and rabbits (q = 0.04, P = 0.8485). The immunized mice with subunit vaccine developed significantly higher anti-rV270 IgG titres than immunized guinea pigs and rabbits (q value was 27.10 and 19.49, respectively, P < 0.01), and there were no anti-rV270 IgG titre differences between the immunized guinea pigs and rabbits with the subunit vaccine (q = 0.25, P = 0.6187). The immunized mice with EV76 elicited higher anti-F1 IgG titres than immunized guinea pigs and rabbits (q value was 40.67 and 29.10, respectively, P < 0.01), whereas there was no difference of F1 IgG titer between immunized guinea pigs and rabbits (q = 0.06, P = 0.8098). The immunized mice, guinea pigs and rabbits with subunit vaccine provided 100% (10/10), 86% (12/14) and 100% (5/5) protection against 10(6) CFU Y. pestis of challenge, respectively. The immunized mice, guinea pigs and rabbits with EV76 vaccine gave 100% (6/6), 93% (13/14) and 100% (6/6) protection against 10(6) CFU Y. pestis of challenge respectively.

CONCLUSIONBALB/c mice is the best small animal model for valuation of protective efficacy of plague subunit vaccine. The guinea pigs showed a high individual variation for this purpose. The rabbits can be used as an alternative model for evaluating plague subunit vaccine.

Animals ; Antibodies, Bacterial ; blood ; Dose-Response Relationship, Immunologic ; Female ; Guinea Pigs ; Immunization ; Immunoglobulin G ; blood ; Mice ; Mice, Inbred BALB C ; Models, Animal ; Plague ; prevention & control ; Plague Vaccine ; immunology ; Rabbits ; Vaccines, Subunit ; immunology

OBJECTIVETo evaluate the efficacy and safety of long pulse 1064 nm Nd:YAG laser therapy in the treatment of onychomycosis of the toenails.

METHODSA total of 104 patients with onychomycosis (461 toenails) were divided by age into ≥60 years group and <60 years group, and each group was further divided into subgroups according to Scoring Clinical Index of Onychomycosis (SCIO) scoring and the location of the compromised toenails. All the toenails were treated with 10 to12 sessions of long pulse 1064 nm Nd:YAG laser therapy at the interval of 1 week. All the patients were followed up for 48 weeks after the initial treatment to assess the clinical efficacy and adverse reactions.

RESULTSThe overall clinical response rate in these patients was 72.5% by the end of the 48-week follow-up. In patients aged <60 years, the clinical response rate and mycological cure rate were significantly higher than the rates in patients aged ≥60 years (P<0.05). No significant differences were observed in the response rates between different SCIO subgroups (P>0.05); the 2nd to 4th toenails showed better outcomes after the therapy than the 1st and 5th toenails (P<0.05). No adverse reactions related with the therapy were recorded in these patients.

CONCLUSIONLong pulse 1064 nm Nd:YAG laser is an effective and safe approach for treatment of onychomycosis of the toenails.

Humans ; Lasers, Solid-State ; Middle Aged ; Nails ; microbiology ; Onychomycosis ; therapy ; Treatment Outcome