Objective:To investigate the correlation of the emm genotypes and virulence genes with the isolation sites of Group A Streptococcus (GAS). Methods:It was a retrospective study.The specimens were collected from children with impetigo in Beijing Children′s Hospital, Capital Medical University from 2006 to 2008 for GAS isolation and identification.A total of 24 GAS strains were isolated from 16 children with impetigo, among which 7 pairs of strains were isolated from the throat and skin of 7 children, and 1 pair of strains was isolated from the vulva and skin of one child, and the remaining 8 GAS strains were isolated from the skin pus samples of 8 children.Polymerase chain reaction was applied to detect the emm genotypes and 13 virulence genes ( speA, speB, speC, speF, speG, speH, speI, speJ, speK, speL, speM, smeZ and ssa). The correlation of the emm genotypes and virulence genes with the isolation sites of GAS strains was analyzed. Results:In this study, four emm genotypes were detected, including emm1.0 (15/24), emm12.0 (4/24), emm22.0 (2/24) and emm160.0 (1/24), and one subtype emm12.19 (2/24) was detected as well.The carrying rates of 13 virulence genes speA, speB, speC, speF, speG, speH, speI, speJ, speK, speL, speM, smeZ and ssa were 58.3%, 100%, 91.7%, 100%, 50.0%, 12.5%, 54.2%, 66.7%, 16.7%, 25.0%, 12.5%, 100% and 91.7%, respectively.All strains carried 5 to 11 virulence genes and they all carried speB, speF and smeZ.There were significant differences in the carrying rate of speA and speJ among the strains with different emm genotypes (all P<0.05). There was no significant difference in the distribution of virulence genes between skin isolates and pharyngeal isolates, including the 5 pairs of strains carrying the emm1.0 genotype (all P>0.05). Conclusions:The distribution of virulence gene of GAS in children with impetigo is significantly correlated with the emm genotype, rather than the isolation site.

Group A Streptococcus (GAS) is a very important pathogen, especially for children.On a global scale, GAS is an important cause of morbidity and mortality.But the burden of disease caused by GAS is still unknown in China and also has not obtained enough attention.For this purpose, the expert consensus is comprehensively described in diagnosis, treatment and prevention of GAS diseases in children, covering related aspects of pneumology, infectiology, immunology, microbiology, cardiology, nephrology, critical care medicine and preventive medicine.Accordingly, the consensus document was intended to improve management strategies of GAS disease in Chinese children.

OBJECTIVE：To study the effects of Modified liuwei dihuang decoction on kidney/bone injury of chronic kidney disease-mineral and bone disorder（CKD-MBD）model rats. METHODS ：The male SD rats were randomly divided into normal group（n＝10），high phosphorus group （n＝30），model group （n＝30），calcitriol group （positive control ，0.09 μg/kg，n＝30）， Modified liuwei dihuang decoction group （10 g/kg by crude drug ，n＝30）. CKD-MBD model was established by high phosphorus and adenine diet for 6 weeks. After modeling ，normal group and model group were given normal diet/high phosphorus diet and intragastric administration of water. Administration groups were fed with normal diet and given corresponding solution intragastrically（water as solvent ），0.1 mL/kg，once a day ，for consecutive 6 weeks. Blood sample of rats in the normal group were collected ，and they were sacrificed after the last administration. Blood sample of 10 rats in each other group were collected ， and they were sacrificed at 2，4 and 6 weeks after administration. The contents of blood urea nitrogen （BUN），serum creatinine （Scr），calcium，phosphorus，iPTH，FGF-23，RANKL and osteocalcin in serum were detected in each group. The bone mineral density（BMD）of femoral was measured ，the morphological changes of renal tissue and bone tissue were observed ，and the percentage of renal tubular injury and the score of renal interstitial fibrosis were calculated. RESULTS ：Compared with normal group，above indexes in high phosphorus group had no significant change at different time points （P＞0.05）. There was no abnormal change in renal/bone tissue. Compared with high phosphorus group at the same time point ，the contents of BUN ，Scr， phosphorus，iPTH，FGF-23，RANKL and osteocalcin in serum ，the percentage of renal tubular injury and the score of renal interstitial fibrosis in the model group were significantly increased ，while the contents of calcium in serum and the BMD of femoral were significantly decreased （P＜0.05 or P＜0.01）. The renal tissue showed diffuse fibrosis. The width of trabecular bone was increased and the number of osteoblasts was decreased. Compared with the model group at the same time point ，the contents of BUN（except for Modified liuwei dihuang decoction group after 2 weeks of administration ），Scr，serum phosphorus ，iPTH， FGF-23，RANKL and osteocalcin ，the percentage of renal tubular injury and the score of renal interstitial fibrosis in Modified liuwei dihuang decoction group and calcitriol group were decreased significantly at each time point ；serum calcium content and BMD（except for 2 weeks of administration ）were significantly increased （P＜0.05 or P＜0.01），and the pathological changes of renal/bone tissue were significantly improved ；there was no statistical significance in above indexes between Modified liuwei dihuang decoction group and calcitriol group （P＞0.05）. CONCLUSIONS ：Modified liuwei dihuang decoction can improve kidney/ bone injury of CKD-MBD model rats ，and improve BMD and regulate disorder of calcium and phosphorus metabolism.

Objective To investigate the feasibility and application value of total enteroscopy with double balloon enteroscopy(DBE)for diagnosis and treatment of patients with obscure gastrointestinal bleeding(OGIB). Methods The clinical and endoscopic data of patients underwent DBE for OGIB in the Department of Gastroenterology,Remin Hospital of Wuhan University from January 2010 to December 2015 were retrospectively analyzed. Results Total enteroscopy was indicated in 36.3%(136/375)of patients. The success rate was 86.0%(117/136)and complication rate was 1.5%(2/136). Negative findings,non-small bowel lesions and small bowel lesions were detected in 44.4%(52/117), 6.8%(8/117), and 48.7%(57/117)of patients with total enteroscopy. Re-bleeding occurred in 8.9%(4/45)of patients with negative total enteroscopy,while 1 small bowel mesenchymoma and 1 gastric fundus Dieulafoy′s lesion were revealed subsequently. Re-bleeding occurred in 33.3%(5/15)of patients with incomplete enteroscopy,and 1 small bowel polyp and 1 small bowel angiectasis were revealed subsequently. Conclusion DBE can complete total enteroscopy within one day and provide important clinical information of OGIB. Non-small bowel lesions,small bowel lesion missed by DBE and potential bleeding lesions in small bowel beyond the reach of DBE should be considered in patients with negative enteroscopy.

Objective To assess the value of double balloon enteroscopy (DBE) in etiological diagnosis of small bowel ulcer.Methods The clinical and endoscopic data of patients undergoing DBE at the Department of Gastroenterology, Renmin Hospital of Wuhan University from January 2010 to January 2016 were reviewed.For patients with small bowel ulcer revealed by DBE, the etiologic diagnosis was retrospectively analyzed.Results One hundred and three patients with small bowel ulcer were included in the analysis.The initial diagnosis based on morphology of ulcer was Crohn's disease (58.3%, 60/103), intestinal tuberculosis (18.4%,19/103), etiology unknown (24.3%, 25/103).The final diagnosis comprehensively based on pathology, clinical data and disease outcome was Crohn's disease (44.7%,46/103), intestinal tuberculosis (18.4%,19/103), lymphoma (3.8%,4/103), eosinophils enteritis (1.9%,2/103), non-steroidal anti-inflammatory drug enteritis (1.9%,2/103), adenocarcinoma (1.0%,1/103), stroma tumor (1.0%,1/103),ischemic enteritis (1.0%,1/103), vascular malformation (1.0%,1/103), Henoch-Schonlein purpura (1.0%,1/103), etiology unknown (24.3,25/103).The accurate diagnosis rate and misdiagnosis rate for Crohn's disease by DBE were 73.3%(44/60)and 10.0%(6/60).The accurate diagnosis rate and misdiagnosis rate for intestinal tuberculosis by DBE were 14/18 and 2/18.Conclusion There are diversity and complexity in etiologic diagnosis of small bowel ulcer.The diagnosis should be based on the comprehensive analysis of morphology of ulcer, endoscopic and surgical pathology and clinical data.Follow up of small bowel ulcer by DBE may provide important information for etiologic diagnosis.

Objective To investigate the role of microRNA-490-5p (miR-490-5p) in the regulation of visceral sensitivity in rats with intestinal dysfunction.Methods The lentivirus vector system was used to construct the rno-miRNA-490-5p lentivirus expression vector.The rats were divided into normal group,diarrhea-predominant irritable bowel syndrome (IBS-D) group,lentivirus empty vector group and the siRNA silent group and the latter three groups were model groups.The efficiency of siRNA was measured by real-time polymerase chain reaction (PCR).The rats were gavaged with 10％ India ink,and then the time of first black stool,water content of feces and threshold of expansion capacity caused abdominal elevation or back arching were calculated.The visceral sensitivity of rats after miRNA-490-5p silenced was evaluated with abdominal withdrawal reflex (AWR) score by stimulating with different intensities of colonic dilatation.The abdominal electrical activity of rats stimulated by colonic distension was measured by BL-420F biological and functional experimental system.The change of the tension of rats isolated colon intestinal stimulated with acetylcholine chloride was also detected by BL-420F biological and functional experimental system.T test was used to compare the differences between the model groups and the normal group.One way analysis of variance was performed for multi-group comparison after miRNA-490-5p interfered.For comparison between two groups among multiple groups,least significant difference (LSD) method was used when the variance was equal,and Games-Howell method was used when the variance was unequal.Results The gastrointestinal propulsion time and the threshold of expansion capacity caused abdominal elevation or back arching of model groups were both lower than those of the normal group ((8.54±4.07) hvs (12.33±2.23) h,(0.56±0.08) mL vs (0.84±0.09) mL),and the differences were statistically significant (t =2.62 and 6.37,both P ＜ 0.05).After distension with 0.8 mL and 1.2 mL sodium chloride solution,the AWR scores of model groups were significantly higher than those of the normal group (3.20±0.56 vs 1.20±0.45,3.73±0.46 vs 2.60±0.55),and the differences were statistically significant (t=7.20 and 4.58,both P＜0.01).There was no significant difference in AWR score between the model groups and the normal group when distended with 1.6 mL sodium chloride solution (3.93 ±0.26 vs 3.80 ± 0.45) (P＞0.05).After miRNA-490-5p silenced,gastrointestinal propulsion time of normal group,IBS-D group,lentivirus empty vector group and the siRNA silent group was (11.12±1.01) h,(6.23±3.17) h,(6.09 ± 2.26) h and (12.36±1.97) h,and the differences among four groups were statistically significant (F=10.55,P＜0.01).The abdominal electrical activity of normal group,IBS-D group,lentivirus empty vector group and the siRNA silent group distension stimulated with 0.8 mL and 1.2 mL sodium chloride solution was (64.91 ± 10.50),(101.79 ±11.73),(80.49±1.27),(66.92±3.24) μV,and (105.09±52.40),(131.71± 16.74),(111.00±6.41) and (95.49± 4.2) μV,and the differences among four groups were statistically significant (F=16.82 and 9.14,both P＜0.05).There was no significant difference in abdominal electrical activity amplitude between silenced group and normal group ((66.92±3.24) μV vs (64.91±10.49) μV and (95.49±4.22) μV vs (105.09±2.40) μV) (all P＞ 0.05).After distension with 1.6 mL sodium chloride solution,the abdominal electrical activity amplitudeof silenced group was lower than the other groups,and the differences were statistically significant (F=11.09,P＜0.01).After adding 1∶1 000 acetylcholine chloride added,the tension of colon of normal group,IBS-D group,lentivirus empty vector group and the siRNA silent group increased by 0.71 ± 0.21,0.81±0.06,0.88±0.21 and 0.43±0.07,however there was no significant difference among the four groups (F=2.57,P =0.100).Conclusions Visceral hypersensitivity existed in rats with intestinal dysfunction.miRNA-490-5p may be involved in the regulation of visceral sensitivity.

BACKGROUND:Previous studies on posterior cervical single door and double door laminoplasty for repair of multilevel cervical myelopathy mainly focus onneurological function and clinical parameters and lack of certain comprehensiveness.
OBJECTIVE:To explore the effects of posterior cervical single door and double door laminoplasty for repair of multilevel cervical myelopathy.
METHODS:We selected 120 patients with multilevel cervical myelopathy and randomly divided into single door group (n=60) and double door group (n=60). The single door group underwent single door laminoplasty. The double door group underwent double door laminoplasty. The blood loss,length of stay, complication rate, neurological function improvement, motion range of cervical vertebrae and imaging changes were compared between the two groups.
RESULTS AND CONCLUSION:(1) Blood loss was significantly less, length of stay was significantly shorter, and the incidence of axial symptom was significantly reduced in the double door group than in the single door group (alP< 0.05). (2) No significant difference in complication rate such as side leakage of cerebrospinal fluid, hematoma, infection and paralysis was detected between the two groups (P> 0.05). The incidence of axial symptoms was significantly less in the double door group than in the single door group (P< 0.05). (3) There were no significant differences in preoperative Japanese Orthopaedic Association scores, postoperative Japanese Orthopaedic Association scores, and improvement in neurological function between the two groups (P> 0.05). Postoperative Japanese Orthopaedic Association scores were significantly increased as compared with that preoperatively in both groups (P<0.05). (4) No significant difference in motion range of cervical vertebrae, inflexion, extension angle and sagittal diameter of spinal canal was detectable between the two groups (P> 0.05). Motion range of cervical vertebrae, extension angle, and inflexion angle were smaler after treatment compared with that preoperatively in both groups (P< 0.05). The increased degree of sagittal diameter of spinal canal was smaler, and the motion range of cervical vertebrae kept better in the double door group than in the single door group. (5) These results indicate that the effect of double door laminoplasty for repair of multilevel cervical myelopathy was significant. The double door laminoplasty can shorten the length of stay, reduce blood loss, axial symptoms, and loss rate of motion range of the cervical vertebrae. In the clinic, treatment can be carried out according to different indications.

Objective To investigate the safety,diagnostic value and clinical impact of double balloon enteroscope (DBE) in elderly patients (aged ≥75 years).Methods Clinical data and DBE findings of elderly patients in the Department of Gastroenterology in Remin Hospital of Wuhan University from January 2010 to January 2013 were retrospectively analyzed and compared with those of younger patients.Results The diagnostic rate,complication rate and clinical impact rate were 69.0％ (29/42),0％ (0/42) and 76.2％ (32/42) in elderly patients.There were no statistically significant differences on those parameters between elderly patients and younger patients (all P＞0.05).For elderly patients,the leading causes of obscure gastrointestinal bleeding (OGIB) were tumor (30.0 ％,9/30) and ulcer (16.7 ％,5/30),while angiectasis (6.7 ％,2/30) was uncommon.Conclusions DBE is an effective and safe method for diagnosis of small bowel disease in elderly patients.Tumor is the most common cause of OGIB,while angiectasis was uncommon.

Objective To investigate the effects of motilin on the voltage dependent potassium channel and L-type calcium channel currents in rat proximal colon smooth muscle cells (PCSM) and to explore its mechanism in increasing colonic motility.Methods PCSM were isolated by collagenase.The voltage dependent potassium channel transit outward current (IKA ) and delayed rectifier current (IKdr) and L-type calcium currents (ICa(L)) were measured by whole cell patch clamp technique.Groups were analyzed by paired t-test.Results There was no significant effect of motilin on IKA and IKdr.L-type calcium channel was dose-dependently activated by motilin from 0.5 × 105 mmol/L to 10.0 ×10-5 mmol/L.At 6 × 10-5 mmol/L motilin and under - 10,0 and 10 mV stimulating voltage,maximum current density increased by 154.61％,62.69％ and 21.02％ respectively and activation kinetics curve obviously left shifted.Half activation voltage decreased from (2.740±1.211) mV prior administration to ( - 25.290 ± 0.614) mV (t =8.534,P =0.007 ) and there was no significant difference in slope factor. Conclusions Motilin increases colonic smooth muscle contraction by promoting calcium influx. However the frequency of colonic smooth muscle contraction could not change with frequency of equilibrium potential and action potential of colonic smooth muscle.

Aim To explore the effect of microRNAs on the differentiation of 3T3-L1 adipocytes and the expression of adipo-related gene-fatty acid binding protein during the adipocyte differentiation.Methods adipo-related microRNAs during 3T3-L1 adipocyte differentiation were screened and identified by micorRNA microarray.Constructed high-expression plasmids of the adipo-related microRNAs,were transfected into the 3T3-L1 preadipocytes by lipofectamine.While the effect of adipo-related microRNAs on the course of 3T3-L1 adipocyte differentiation was observed,the protein and mRNA expression level of fatty acid binding protein(FABP4)were analyzed by Western blot and RT-PCR during 3T3-L1 adipocyte differentiation.Results The expression profiles of microRNAs have significant changed during 3T3-L1 adipocyte differentiation,in which 35 microRNAs among them down-relation,the most lowly expression is miR-24;17 microRNAs among them up-relation,the most highly expression is miR-21.MiR-24 significantly inhibited adipocyte differentiation and maturity,while miR-21 have no significant effect.MiR-24 significantly inhibited the expression of FABP4,but had no effect on the level of its mRNA;miR-21 had no effect on the expression of protein and mRNA of FABP4.Conclusion There exist adipogenic-related microRNAs during 3T3-L1 adipocyte differentiation; miR-24 play an important role in the regulation of 3T3-L1 preadipocyte differentiation into adipocyte and the(FABP4)protein expression.