Aim: To clone and over-express the gene encoding aminoglycoside(AG)phosphotransferase(APH)from clinical MRSA isolates in E.coli and to develop an assay method for the recombinant APH.Methods: The susceptibility of clinical MRSA isolates to AGs was tested by disk diffusion.A nucleic acid sequence encoding APH was amplified from the genomic DNA of an isolate and ligated to expression vector pET-28a,and then trans-formed into E.coli BL21(DE3).After purification of the recombinant protein by affinity chromatography,the phosphorylation activity of the enzyme was determined by ESI-MS and disk diffusion.Flesults: All 6 clinical MRSA isolates were unsusceptible to AGs.After cloning and expression,the recombinant APH was purified to90%.The in vitro activity assay indicated that the recombinant protein could inactivate kanamycin B in the assay mixture within 2 h.Conclusion: The recombinant APH showed excellent enzymatic activity.The assay method was simple and convenient,which may provide the basis of developing a screening model for APH inhibitors.

Objective To observe the effect of acupoint application of Chinese medicine on preventing constipation after stroke. Methods By means of randomized, double-blind, controlled study design, 96 patients of yang-syndrome stroke were randomly divided into observation group and control group, 48 cases in each group. Both groups were given conventional treatment for stroke and routine nursing for preventing constipation, and additionally, the control group was given acupoint application of placebo while the observation group was given acupoint application of self-made umbilical paste of Radix et Rhizoma Rhei(Dahuang) and Natrii Sulfas (Mangxiao). The incidence of constipation, frequency of laxative usage and adverse reactions of the two groups were observed. Results (1) The incidence of constipation of the observation group was lower than the control group on treatment day 7 and 14 , the difference being statistical significant(P < 0.05 or P < 0.01).(2)On treatment day 7 and 14, the Bristol scores of stool properties of observation group were increased compared with those before treatment(P < 0.05), but the changes of scores in the control group were insignificant(P > 0.05). The observation group had better effect on improving stool properties(P < 0.05).(3) The frequency of use of laxatives was reduced in the observation group compared with the control group, the difference being significant(P<0.05).(4) The incidence of adverse reaction was 6.25%in the control group and was 8.333%in the observation group, the difference being insignificant (P>0.05). The adverse reaction of both groups was mild, and had no effect on the accomplishment of the treatment. Conclusion Acupoint application of Chinese medicine is effective and safe on preventing constipation after stroke, showing good prospects for clinical application.

Background Corneal biomechanical properties is important in the safety assessment of corneal refractive surgery.Corvis is a new device for measuring corneal biomechanics properties.Objective This study was to observe the correlation among corneal thickness, Corvis intraocular pressure and corneal biomechanical properties with Corvis.Methods A prospective observational study was performed.One hundred and fifty eyes of 75 patients with corneal thickness from 501 μm to 590 μm were divided into three groups according to the corneal thickness:low corneal thickness group (corneal thickness range from 501 μm to 530 μm), middle corneal thickness group (corneal thickness range from 531 μm to 560 μm) ,and high corneal thickness group (corneal thickness range from 561 μm to 590 μm);and 50 eyes of 25 patients for each group.The difference of intraocular pressure, corneal thickness and deformation amplitude (DA) among the three groups were analyzed by one-way ANOVA and the correlation among the groups were analyzed by liner regression.Results The DA in the low corneal thickness group and middle corneal thickness group were significantly higher than that in the high corneal thickness group (P ＜ 0.05).The intraocular pressure was statistically different among the 3 groups (F =9.98, P＜0.05).DA was negatively correlated with intraocular pressure and corneal thickness (r=-0.84,-0.33;both at P＜0.01), with the linear regression DA =1.69-0.04×IOP (F=366.19, t=-19.14,P＜0.01).Conclusions Corneal thickness cannot simply represent the corneal biomechanical properties in the safety assessment of corneal refractive surgery,IOP should be considered.

Objective To investigate the effect of ultraviolet ray on DNA methylation in systemic lupus erythematosus (SLE) and its role in the pathogenesis of SLE.Methods Peripheral blood mononuclear cells (PBMC) were isolated from 45 SLE patients and 20 healthy controls and were cultured in vitro.Then they were irradiated with 311 nm narrow band-ultraviolet my (NB-UVB) in various dosages.DNA methylation was,detected by high-performance capillary electrophoresis and DNA methyltransferase 1 (DNMT1) mRNA expression was measured by real-time quantitative PCR.Results The level of DNA methylation in SLE pafients was lower than that in the control group (P=0.014).DNA methylation was,decreased after UVB exposure with the dosage of 50,100 mJ/cm2 (P＜0.01),but no significant difference could be found in the DNMT1 mRNA expression.No significant correlation was found between the level of DNA methylation and systemic lupus erythematosus activity index (SLEDAI).Conclusion Changes take palce in DNA methylation after UVB exposure,and UVB may affect the pathogenesis of SLE by changing DNA methylation.

Objective: To observe the clinical efficacy of acupuncture plus bloodletting therapy in treating insomnia in people with blood stasis constitution. Methods: Seventy-two patients were randomized into a treatment group and a control group by using the random number table, 36 cases each. The treatment group received acupuncture plus bloodletting therapy, while the control group was intervened by oral administration of estazolam. One-week treatment was taken as a course, for 3 successive courses in total. A follow-up study was conducted 3 months later. The Pittsburgh sleep quality index (PSQI) was evaluated before and after the intervention, as well as in the follow-up. The clinical efficacies were also compared. Results: The total effective rate was 88.9% in the treatment group versus 83.3% in the control group, and the between-group difference was statistically significant (P<0.05). After the intervention, the PSQI scores dropped significantly in both groups (both P<0.05); the between-group difference in PSQI score was statistically significant (P<0.05). The follow-up study showed that the between-group difference in the global score of PSQI was statistically significant (P<0.05). Conclusion: Acupuncture plus bloodletting therapy can produce a more significant efficacy than oral administration of estazolam in treating insomnia in people with blood stasis constitution.

Objective: To dynamically study the use of antiseptic drugs at tertiary public hospitals in the localities for general information. Methods: In this paper, the panel data were analyzed using dynamic monitoring data from the " monitoring network for clinical application of antiseptic drugs" . Such means as descriptive statistical analysis, single-factor analysis and logistic regression analysis were used to learn the use and monitoring of such drugs. Results: We found that the use of such drugs at tertiary hospitals has improved significantly nationwide in recent years, yet with some gaps up to the standard. Given such progress, the economic and social burden caused by the unreasonable use of antiseptic drugs calls for more attention. Conclusions It is imperative to strengthen the supervision of such drugs and promote their rational use so as to reduce the social and economic burden.

BACKGROUNDIt has been known that the growth of solid tumors is dependent on angiogenesis, and neoangiogeneses of tumor become main target to control tumor growth. The aims of this study are to investigate the inhibition effect of replicate-deficient adenovirus encoding the soluble form of mouse vascular endothelial growth factor receptor 1 (sFlt1-Adv) on angiogenesis and tumor growth in established tumor model.

METHODSMouse Lewis lung cancer cells were inoculated subcutaneously into C57 mice. sFlt1-Adv, GFP-Adv and normal saline were injected twice intravenously after establishing Lewis cancer model. Diameters of tumors were measured every other day. Tumors were resected, weighed and fixed in 3% paraformadehyde. Microvessel density of tumors was determined by immunohistochemical staining with anti-CD31 antibody.

RESULTSThe planted tumor volume and weight in sFlt1-Adv group were significantly lower compared with the two controls (P < 0.01). Its inhibition rate was 71.8%. The microvessel density in sFlt1-Adv group decreased markedly compared with that of the control groups (P < 0.01).

CONCLUSIONSsFlt1-Adv can inhibit the growth of tumor through the inhibition of tumor angiogenesis. sFlt1-Adv may be potentially valuable for clinical treatment of solid tumor.

AIMTo investigate the protective effect of exogenous carbon monoxide (CO) on the liver injury induced by ischemia/reperfusion (I/R) of hind limbs in rats.

METHODS100 SD rats were divided randomly into sham operated group (S), S+ CO group (SC), I/R group (I/R), I/ R+ CO group (RC). A rat model of ischemia in hind limbs and the reperfusion liver injury was established with the occlusion of the femoral arteries for 4 h and re-opening for 6 - 72 h, 10 d. The rats in SC and RC groups were exposed to air containing CO (the volume traction of CO: 0.05%) for 2 h before and after reperfusion or the corresponding control time point, while the other two groups were exposed to the routine air. The pathologic changes of liver tissue were morphologically observed by HE stain. Serum GPT activity was tested by Automatic Biochemical Analyzer. The percentage of apoptosis, expression levels of bax and bcl-2 protein in liver tissue were detected by Flow Cytometry.

RESULTSThere was no difference between S and SC groups. Compared with SC group: (1) Pathological changes in liver tissue were significant in I/R and RC groups. (2) The serum GPT activity of I/R and RC groups was obviously increased. (3) In IR and RC groups, the percentage of apoptosis in liver tissue was all significantly increased. (4) The bax expression level was significantly increased. Compared RC group with I/R group: (1) Pathological change was slight. (2) The serum GPT activity was depressed. (3) The percentage of apoptosis and expression level of bax protein in liver tissue were depressed. (4) The expression level of bcl-2 protein in liver tissue was increased.

CONCLUSIONExogenous CO could attenuate liver tissue injury induced by limbs I/R in rats.

Animals ; Carbon Monoxide ; pharmacology ; Extremities ; blood supply ; Female ; Liver ; blood supply ; pathology ; Liver Diseases ; etiology ; pathology ; prevention & control ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; complications ; prevention & control

OBJECTIVETo investigate the therapeutic effect of island flap pedicled with dorsal cutaneous branches of digital internal artery for skin defect at the same finger.

METHODSFrom June 2005 to June 2008, 52 fingers in 45 cases with skin defects were treated with island flap pedicled with dorsal cutaneous branches of digital internal artery for skin defect at the same finger.

RESULTSPartial necrosis happened at the distal end of 4 island flaps. All the other flaps survived completely. The patients were followed up for 4-8 months with satisfactory cosmetic and functional results. The two-points discrimination distance was 6. 0-9.0 mm (average, 7.4 mm) in flaps with nerve anastomosis, and 8-10 mm (average, 9 mm) in flaps without nerve anastomosis. Minor pigmentation occurred in 5 fingers, cicatricial contracture around the skin graft in 4 fingers, and pain in donor site scar in 2 fingers. The hand function was assessed as excellent in 42 fingers, as good in 7 fingers and medium in 3 fingers.

CONCLUSIONSThe main artery will not be sacrificed when the island flap is used. It is ideal for the treatment of finger skin defect.

Adolescent ; Adult ; Female ; Finger Injuries ; surgery ; Fingers ; blood supply ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Skin ; injuries ; Skin Transplantation ; methods ; Surgical Flaps ; Treatment Outcome ; Young Adult

Objective To find out factors related to avascular necrosis (AVN) of femoral head through a retrospective study of patients with femoral neck fractures treated with cannulated screws.Methods Two hundred and thirty cases of femoral neck fracture whn had been treated with cannulated screws in our department from August 1994 to October 2003 were analyzed statistically.Six factors of patient,age,gender,fracture type (Garden clas- sification),reduction method (close or open),the quality of reduction (Garden Index) and time interval from injury to surgery were analyzed statistically with rank sum test and logistic multiple regression.Results Factors that might have an impact on avascular necrosis of femoral head were ranked in the following likelihood sequence: fracture type,quality of reduction,reduction method,time interval from injury to surgery and age.Gender was found to have little impact on the prognosis of femoral neck fracture (P＞0.05).Conclusions Fracture displacement is the most critical factor that influences the prognosis.Anatomical reduction (close or open) must be achieved to re- duce the risk of AVN.Timing of surgery shows its importance.High age will not lead to an increased incidence of AVN.