Objective Methicillin-resistant Staphylococcus aureus (MRSA) is a growing public health concern that has been associated with pediatric fatalities. This study investigated the genotypes of staphylococcal cassette chromosomal mec (SCCmec) and Panton-Valentine Leukocidin (PVL) in MRSA strains isolated from Shanghai Children's Hospital by PCR. Methods A total of 30 strains of MRSA were isolated from various clinical specimens from October 2005 to June 2006. The antimicrobial susceptibility was measured by agar diffusion method. SCCmec typing was conducted using a novel multiplex PCR assay allowing for concomitant detection of methicillin resistance (mecA gene) to facilitate detection and classification of all currently described SCCmec typesⅠ, Ⅱ, Ⅲ, Ⅳa, b, c, d andⅤ. PVL gene was also determined by PCR. Results mecA gene was positive in all the strains. SCCmecⅡ was identified in 6(20.0%) isolates, SCCmecⅢ in 15(50.0%) isolates, SCCmecⅤ in 2 and SCCmecⅣa in 1 isolate. Six MRSA strains were non-typeable. The isolates with SCCmecⅡ or SCCmecⅢ were resistant to multiple antibiotics. The strains harboring SCCmecⅣa or SCCmecⅤwere susceptible to all antibiotics except β-lactams. Eleven (36.7%) isolates were PVL positive. The genotypes and subgenotypes of staphylococcal chromosomal cassette mec of eleven PVL-positive MRSA were SCCmecⅡ(1 isolates), SCCmecⅢ (5 isolates), SCCmecⅣa (1 isolate), SCCmecⅤ (2 iso-Lates) non-typeable (2 isolates). Conclusions SCCmecⅡ and SCCmecⅢ are the major genotypes of MRSA in our hospital. These isolates are multi-resistant to antibiotics. The prevalence of PVL gene is higher in SCCmecⅡ- or SCCmeⅢ-positive MRSA. The isolates with SCCmecⅡ or SCCmecⅢ were resistant to multiple antibiotics.

Objective:To observe the clinical efficacy of electroacupuncture (EA) combined with tuina for chronic tension-type headache (CTTH). Methods:A total of 97 CTTH cases were randomly allocated into an observation group (n=52) and a control group (n=45). Patients in the observation group were treated with EA plus tuina based on pattern identification, whereas patients in the control group were treated with oral amitriptyline and oryzanol. Patients in both groups were treated for 8 weeks. A follow-up was conducted 3 months after the treatment. The intensity, duration, and frequency of the headache were recorded and compared before and after the treatment. Additionally, the patients' psychological state and quality of life (QOL) were compared between the two groups. Results:There were intra-group statistically significant differences in headache intensity score, headache duration, and headache frequency after the treatment and during the follow-up compared with those before the treatment (allP<0.05); and there were between-group statistically significant differences during the same time frame (allP<0.05). The Hamilton depression scale-17 items (HAMD-17) and Hamilton anxiety scale (HAMA) scores were significantly reduced in both groups after the treatment or during the follow-up (allP<0.01); and there were no between-group statistical significances during the same time frame (allP>0.05). The World Health Organization quality of life-BREF (WHOQOL-BREF) scores were significantly reduced in both groups after the treatment or during the follow-up (allP<0.05); and the scores in the observation group were significantly lower than those in the control group (allP<0.05) during the same time frame. The total effective rate was 88.0% in the observation group, versus 71.4% in the control group, showing a significant difference (P<0.05). Conclusion:EA combined with tuina can significantly decrease the frequency of chronic tension-type headache, alleviate headache intensity, shorten headache duration, and improve the patients' wellness. It is better than amitriptyline plus oryzanol.

Objective To investigate the association between carotid atherosclerosis(AS) and serum magnesium(Mg) in hemodialyzed (HD) patients.Methods Clinical index was measured,and intimamedia thickness (IMT) of extracranial common carotid artery and presence of atherosclerotic plaques were determined by high-resolution B-mode ultrasonography.The data were analyzed between plaque positive group and plaque negative group.Results The age,serum phosphate (P),total cholesterol (TCH),low density lipoprotein (LDL),serum C-reactive protein (CRP),serum albumin(Alb),and serum Mg all had significant difference between two groups(t =4.153,2.908,2.301,6.322,5.791,2.341,7.778,P ＜0.01 or P ＜ 0.05).The risk factors of HD patients with AS were serum Mg and CRP(P ＜ 0.01).Conclusions The occurrence of AS was related to low Mg and high CRP in HD patients.

Objective To investigate the role of cHSP60 in the pathogenesis of murine cervicitis.Methods Fifty female C3H/HeN mice were randomly and equally classified into 5 groups, including the control group receiving no treatment and 4 groups receiving intravaginal inoculation of cHSP60 (cHSP60 group),live elementary bodies of Chlamydia trachomatis mouse pneumonitis (MoPn group), inactive elementary bodies of MoPn (inactive MoPn group) and growth medium (medium group), respectively. Five days after the inoculation,cervical tissue was resected from these mice and subjected to pathological examination. Results There were varying degrees of inflammatory reaction characterized by neutrophil infiltration, necrosis and shedding of mucosal cells in the cervices of mice in cGSP60 and MoPn groups. No statistical difference was observed in the incidence of cervicitis (90% vs. 80%, P ＞ 0.05), neutrophile count [76.00 (25.0 - 80.0) vs. 25.00 (8.75 -32.5), P＞ 0.05] or inflammation score [12.5 (11.5 - 14.25) vs. 9.00 (8.00 - 11.5), P ＞ 0.05] between the cHSP60 and MoPn group. The inflammatory reaction was weak with decreased incidence of cervicitis (40%),inflammation score [0.00 (0.00- 12.50)] and neutrophile count [0.00 (0.00- 15.50)] in inactive MoPn group compared with the cHSP60 and MoPn groups (all P ＜ 0.05). A small number of neutrophils migrated into the superficial layer of cervical mucosa in only 2 mice in the medium group. Conclusion cHSP60 may be a primary pathogenic factor in chlamydial genital tract infection.

Objective To explore the application and efficacy of autogenous septal cartilage in the correction of cleft lip nasal deformity.Methods A total of 31 patients with secondary nasal deformity of cleft lip were involved in this study.After autologous nasal cartilage was released,the dislocation of nasal cartilage was corrected,nasal septum cartilage was used as substitute to raise the alar collapse,to reshape the nasal tip cartilage structure and correct the nasal deformity.Results Postoperative incision was primarily healing.For cartilage donor area there was no serious complications,and incision scar was concealed and unobvious; after 6 months to 2 years follow-up,there was no cartilage absorption deformation,the tip of the nose was flat,crack side nosewing collapses,and nasal columella skew was totally corrected,with stable rhinoplasty effects.Conclusions For secondary nasal deformity of cleft lip,the use of nasal septum cartilage to repair nose deformity can not only solve the cartilage source,but also give septal straightening and at the same time the correction effect of external nasal deformity is good.

Objective: Recent studies have suggested that congestive heart failure(CHF) is related to neuroendocrine system and cytokines,and its activation contributes to myocardial remodeling and leads to the development and progression of CHF.Tumour necrosis factor-?(TNF-?) is one of the most important factors.This study intends to establish the animal model of isoproterenol(ISO)-induced CHF in rats and to study the changes of serum TNF-?.Methods:Male SD rats were randomly allocated to the ISO-induced CHF group(n=50) receiving two subcutaneous injections of 170mg ISO per kilogram of body and the healthy control group(n=10) receiving two subcutaneous injections of 0.25ml normal saline.At 18th week,the hemodynamic parameters including HR,LVSP,LVEDP and ?dp/dtmax were assessed.TNF-? in plasma were all detected.Results:Compared with the control group,the LVSP、?dp/dtmax significantly decreased and the HR,LVEDP enlarged notably in CHF group(P

ObjectiveTo observe the ultrastructural changes of Chlamydia trachomatis after treatment with azithromycin.Methods The Chlamydia trachomatis laboratory strain (D/UW-3/Cx) was cultured in McCoy cells with or without the presence of azithromycin of 0.0667,0.1340,0.1900,0.2680 and 0.3330 mg/L for 48 hours.The ultrastructural changes of host cells andChlamydia trachomatis were observed by transmission electron microscopy.ResultsAfter 48-hour culture,vesicles increased in number both inside and outside of the inclusion bodies with the rise in azithromycin concentration; there were abnormally large reticulate bodies,some of which experienced abnormal division and even necrosis or breakdown; the number of elementary bodies was decreased,while their size was enlarged,with a more wrinkled outer membrane.No inclusionbodieswereseenwhentheconcentrationofazithromycinwas0.333mg/L. Conclusions Azithromycin can induce an increment in the outer membrane of Chlamydia trachomatis,formation of vesicles,abnormal enlargement or breakdown of reticulate bodies,and a decrease in elementary bodies.

0.05).Conclusions IL-11 unlikely to stimulate the growth of the CEM leukemia cell.And did not affect the antitumor effect of HD-MTX.It can be used as an assisting drug in program of acute lymphoblastic leukemia(ALL) therapy.

Orobanche caerulescens is an important medicinal resource in Orobanchaceae. The present study aims to establish methods for determination of acteoside, crenatoside, and total phenylethanoid glycosides in O. caerulescens, and determine the content in 15 samples to evaluate the resource utilization of this medicinal plant. The content of acteoside and crenatoside were quantitatively determined by HPLC, while total phenylpropanoid glycosides was estimated by UV-VIS spectrophotometry. According to the results, the content of acteoside was the highest in O. caerulescens, followed by crenatoside. The contents of acteoside, crenatoside, and total phenylethanoid glycosides were between 1.15% - 15.60%, 0.83% - 4.47%, and 6.78% - 27.43%, respectively, which had significant differences. The acquisition time has great influence on the content of main components of O. caerulescens. The content of phenylethanoid glycosides is higher in the samples which were collected at the flowering stage. The two determination methods were proved to be simple, accurate and reliable, and can be used to evaluate the quality and resource utilization of O. caerulescens.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Glycosides ; analysis ; Orobanche ; chemistry

In order to improve the thermostability of the Penicillium expansum Lipase (PEL), the lipase encoding genes was mutated by site-directed mutagenesis. A recombinant vector pAO815-ep8-K55R which contain double mutant genes was constructed by overlap extension PCR using the cDNA of a random-mutant lipase ep8 (a single site mutant) as the template and two special primers were used to generate another mutation site K55R. The recombinant vector was transformed into Pichia pastoris GS115 by electroporation and the recombinant mutant GS-pAO815-ep8- K55R can secret double-mutant lipase PEL-ep8-K55R-GS into the medium when it was induced by Methanol. The yield of the double-mutant lipase is 508 u/mL, which is 81% that of the wild type lipase PEL-GS (627 u/mL) and 55% that of random-mutant PEL-ep8-GS (924 u/mL). The specific activity of double-mutant lipase is 2309.1 u/mg, which is similar to random-mutant lipase PEL-ep8-GS and the wild type lipase PEL-GS. The optimum temperature of the double-mutant lipase is same with the wild type lipase PEL-GS and random-mutant lipase PEL-ep8-GS. While the Tm of the double-mutant lipase is 41.0 degrees C, 2.3 degrees C higher than the wild type lipase PEL-GS and 0.8% higher than the random-mutant lipase PEL-ep8-GS, indicating that the double-mutant lipase PEL-ep8-K55R-GS has higher thermostability.
Electroporation ; Enzyme Stability ; Hot Temperature ; Lipase ; genetics ; metabolism ; Mutagenesis, Site-Directed ; Mutant Proteins ; metabolism ; Penicillium ; enzymology ; Pichia ; genetics ; metabolism ; Protein Engineering ; methods ; Recombinant Proteins ; biosynthesis ; genetics