Objective To investigate molecular typing and drug resistance patterns of 98 Klebsiella pneumoniae (K.pneumoniae) isolated from type 2 diabetes patients complicated with maxillofacial infection,to research the virulence and resistance mechanisms.Methods The study was a prospective study that adopted the method of continuous sampling from fixed location,from March 2010 to October 2012.The maxillofacial surgery patients diagnosed with type 2 diabetes complicated with maxillofacial infection were chosen in 7 hospitals in Zhengzhou as the research object,and a total of 431 pus sample were collected continuously,in which 98 strains K.pneumoniae were isolated and identified.The Kirby-Bauer disk diffusion test was conducted in 98 strains to determine the resistance to 19 antibacterial agents.K.pneumoniae chromosomal DNA were digested by restriction endonuclease Xba Ⅰ and analyzed by pulsed-field gel electrophoresis (PFGE).PFGE patterns of K.pneumoniae strains were analyzed using Fingerprinting software.The relationship between the molecular types and resistance phenotype was observed.The extended spectrum β-1actamase-producing K.pneumoniae were screened out by the double disc synergy test (DDST)Polymerase chain reaction (PCR) was used to detect resistant genotypes,serotype and virulence genes.The purified PCR products of resistant genes were cloned and sequenced.Hypermucoviscosity phenotype of all strains were determined by string test.Results Much severer drug-resistance for K.pneumoniae was identified and the result of extended-spectrum beta-lactamase producing rate was 57.1 ％.Ninety-eight strains were dispatched into 13 groups by PFGE.No dominant bands and specific extended-spectrum beta-lactamase DNA bands were found.The results of PCR showed that among the 56 strains of extended spectrum β-lactamase-producing K.pneumoniae,40 were positive for blaSHV (accounting for 71.4％),28 positive for blaTEM (accounting for 50.5％),21 positive for blaCTX-M (accounting tor 37.5％).The sequencing results were as follows:TEM-1,CTX-M-3 and a variety of SHV.Serotype K1,K2,K3,K5,K20,K54 and K57 and 3 kinds of virulence genes were detected,but not in strong toxicity-based.Hypermucoviscosity positive rate was 31.6％ (31/98).Conclusion Much severer drug resistance of K.pneumoniae in this study was identified and resistant mechanism was complex,in which strong toxicity serotype and virulence geues exist,which need more attention from clinical.

Objective To investigate the relation of the MR manifestations and the pathological characteristics of dissociated discherniation.Methods Characteristics of pathology and MRI of 23 cases of dissociated disc herniation were retrospectively analysed.Results Of 23 cases,the dissociated disc was low signal intensity(compared with spinal cord) on T_1WI and T_2WI with distinct margine in 7 cases,the adhere and inflammation were slight on pathology;iso-or hypo-signal intensity on T_1WI,low signal intensity in the center and slight high signal intensity at periphery on T_2WI with an distinct margine in 13 cases,while the adhere and inflammation were obvious on pathology;on T_1WI slight high signal intensity on T_1WI and iso-intensity on T_2WI in 3 cases,and fibroplasia was showed on patholgy.In the 12 cases with enhanced MRI,the lesions were slight enhancement at periphery and enhancement in the center in 7 cases,no enhancement in 3 cases,slight enhancement in 2 cases.Conclusion MRI signal characteristic can reflect pathologic changes expressly of the dissociated disc herniation.

Objective To evaluate the role of multidetector CT(MDCT) and high magnetic field MRI in diagnosis of small cystic-solid renal mass. Methods Fifty-two cases with small renal cystic-solid mass(≤3 cm) were consecutively collected,including small cystic-solid renal cell carcinoma(n=25),carcinoid(n=1),complex cysts(n=16),small angiomyolipoma(n=7) and benign cystic nephroma(n=3).All were examined by both 1.5T MRI and multidetector CT at intervals between 3 days and 2 months. Results All cases were proved by pathology.Multi-planar reconstruction techniques were useful for MDCT in differentiating small cystic-solid renal mass,with the sensitivity of 98.1%,which was as high as MRI.However,the accuracy for MDCT was 71.2%,significantly lower than that of MRI(90.4%)(P=0.001).MRI helped to identify the components and structure of renal masses,and behaved better in the detection of pseudo-capsule of renal cell carcinoma(57.7%).Conclusion High magnetic field MRI may play an important role in the diagnosis of small renal cystic-solid masses,and it may be feasible as a noninvasive examination when CT can not make the ultimate determination.

Objective To analyze the clinical features of 27 Chinese patients with Kennedy's disease(KD), a sex-linked inheritance disorder. Methods We collected the clinical data of 27 KD patients whose diagnosis were confirmed by gene examination to analyze their clinical features, as well as their serum levels of sex hormones and biochemical indicators. Results Patients with spinal and bulhar muscular atrophy tend to have an adult onset, exhibiting a slow progression of lower motor neuronal weakness and atrophy involving limbs and bulbar zones. Part of the invalids presented signs of androgen insensitivity such as gynecomastia or reduced sexual function, even though their degrees of sex hormones were normal. There was mild motor functional lesion which correlated with the course of the disease ( r = 0. 77, P = 0. 000). The degrees of creatine kinase ( ( 920. 10 ± 495.54 ) U/L ) and triglyceride ( ( 3.27 ± 2. 78) mmool/L) increased mildly and was significantly different from the levels of healthy ones (107.20 U/L,t =7.517,P =0.000;1.40 mmol/L, t =2.687,P =0.017). Conclusions Basically, Chinese KD patients present the same clinical features as the literature reports, however they have distinctive aspects such as elevated degree of triglyceride. The present research can help us to understand the features of Chinese KD more particularly.

OBJECTIVE: To evaluate the efficacy of compound glycyrrhizin in preventing liver injury induced by antituberculosis drugs. METHODS: A total of 180 cases with pulmonary tuberculosis were randomly divided into control group and trial group: 90 cases in the control group were treated with antituberculosis drugs alone for 6～8 months,and another 90 cases in the trial group were treated concomitantly with antituberculosis drugs and compound glycyrrhizin,the incidences of liver injury between 2 groups were monitored. RESULTS: The incidences of liver injury of the control group and the trial group were 32.2% and 8.9%,respectively(P

Objective To investigate the properties of proliferation and differentiation of neonatal rat retinal progenitor cells (RPCs) in vitro. Methods RPCs were isolated from neonatal SD rats neural retina and cultured in DMEM/F12+N2 with EGF and bFGF (suspension medium )or 10%FBS without EGF and bFGF (differentiation medium). The cells grew as suspended spheres or adherent monolayers, depending on different culture conditions. The neural stem cells or retinal progenitors, neurons, astrocytes, retinal ganglion cells, rod photoreceptors and the proliferating cells were evaluated with immunofluorescence analysis by Nestin or Pax6, Map2, GFAP, Thy-1, Rhodopsin and BrdU antibodies respectively. Results RPCs could propagate and differentiate in suspension or differentiation medium and express the markers of Nestin (92.86%) or Pax6 (86.75%), Map2 (38.54%), GFAP (20.93%), Thy-1 (27.66%) and Rhodopsin(13.33%)in suspension medium; however, Nestin (60.27%), Pax6 (52%), Map2 (34.94%), GFAP (38.17%), Thy-1(30.84%) and Rhodopsin (34.67%) in differentiation medium. 96.4% of the population in the neurospheres was BrdU-positive cells. The cells could spontaneously adherent forming some subspheres and retinal specific cell types. Conclusion Neonatal rat RPCs possess the high degree of proliferation and can differentiate into neurons, astrocytes, retinal ganglion cells and rod photoreceptors in vitro. There are different proportions for RPCs to differentiate into specific cell types.

Objective To construct the genetic recombinant adenovirus vector carrying the human growth and differentiation fac-tor-5(GDF-5) gene by using AdEasy-1 adenovirus vector system and to amplify and prepare the recombinant adenovirus in HEK 293 cells .Methods Human GDF-5 gene obtained by PCR was inserted into plasmid pMD19-T and the 1 .7 kb GDF-5 cDNA sequence was cloned into the adenoviral shuttle plasmid pShuttle-cytomegalovirus(CMV) of the AdEasy-1 system .It was identified by DNA sequencing and a digestion with Hind Ⅲ restriction enzyme .The resultant pShuttle-CMV-GDF-5 was used to generate the adenovi-ral GDF-5 vector through homologous recombination with the adenoviral backbone plasmid ,pAdEasy-1 in BJ5183 bacterial cells .It was selected by kanamycin and identified by a digestion with Hind Ⅲ restriction enzyme and amplified in XL10-Gold competent bac-teria .The DNA of recombinant adenovirus vector was finally linearized by Pac Ⅰ and the adenoviral recombinants were used to pro-duce adenoviruses in HEK293 packaging cells ,resulting in an Ad-GDF-5 identified by Western blot .The virus titer was assayed by TCID50 .Results GDF-5 cDNA sequence obtained by PCR was 1 .7 kb .Gene sequencing results indicated that the sequence was i-dentical with the one in GENBANK .Cloned sequence 1 .7 kb(GDF-5) was obtained by a digestion with Hind Ⅲ restriction enzyme after GDF-5 cDNA segment was cloned into pShuttle-CMV and AdEasy-1 .Western blot showed that two bands migrating at ap-proximately 15 and 55 kDa were clearly observed in PVDF membrane .These data confirmed that HEK293 cells expressed a large number of mature GDF-5 protein after infected with Ad-GDF-5 .Our research results demonstrated that recombinant adenovirus vector GDF-5 was successfully constructed .The virus titer was 5 .6 × 109 PFU/mL .Conclusion Recombinant adenovirus vector carrying the human GDF-5 gene is successfully constructed by using the AdEasy-1 adenovirus vector system .Moreover ,the Ad-GDF-5 with high titer is prepared .These provide the basis for further study of the biological function of GDF-5 and the gene thera-py of its related diseases .

Objective To study the effective composition of Aster ageratoides Turcz for relieving asthma and its toxicity . Methods Seventy‐two qualified male guinea pigs ,body mass(300 ± 20)g ,were randomly divided into the control group ,aminophyl‐line group(0 .047 g/kg) and different ratio group of total saponins and total flavonoids (8∶1 ,4∶1 ,2∶1 ,1∶1 ,1∶2 ,1∶4 ,1∶8) . Every group received at dose of 0 .38 g/kg once a day for 3 days by intragastric administration .The effect of relieving asthma was taken as the indicators for the optimization of compositions ,which was detected by observing the latent period of asthma induced by acetylcholine‐histamine in guinea rats .The relaxation percentage of smooth muscle retracted by acetylcholine was detected by Pow‐erlab biological signal collecting system when aminophylline(1 .25 mg/mL) and different matching drugs(0 .02 g/mL) were added in Krebs′solution .The acute toxicities of total saponins ,total flavonoids and mixture of total saponins with total flavonoids (ratio of 1∶2) were detected by classical acute toxicity test of mice .Results Compared with the control group ,aminophylline group ,each group latent period of inducing asthma significantly or very significantly prolonged (P<0 .05) ,among them ,1∶1 ,1∶2 ,1∶4 group and other matching group were significantly different(P<0 .05) ,but 1∶2 group cited the longest incubation period .Aminophylline group and the ratio could significantly relaxe guinea pig tracheal smooth muscle retracted by acetylcholine .Aminophylline group di‐astolic percentage was the largest(24 .12 ± 1 .20)% ,all the groups were less than the percentage of diastolic aminophylline group (P<0 .05) ,but in 1∶2 group with the closest .Aster ageratoides Turcz ,tatarian aster total saponins ,total flavonoids and the ratio of 1∶2 to mice maximum tolerance was respectively 300 ,345 ,325 g/kg .Conclusion For the same total dosage ,the relieving asth‐ma effects of proportions of 1∶2 Aster ageratoides Turcz were obvious .There are no abnormal changes in the acute toxicity trila with total flavonoids ,total saponins ,as well as the ratio of 1∶2 ,safety is relatively good .

Objective To investigate the clinical efficacy of preoperative full reset combined with minimally invasive treatment of extreme distal pilon fractures.Methods A retrospective analysis was made on 34 patients (35 ankles) with tibial fractures extremely close to the distal articular surface treated surgically between January 2011 and January 2015.There were 21 mnales and 13 females,aged 20-71 years (mean,36.2 years).Injury resulted from traffic accidents in 32 patients and high falls in two.Using the AO/OTA fracture classification system,type 43-B3 was noted in three patients,43-C1 in five patients,43-C2 in 18 patients and 43-C3 in eight patients.Calcaneal traction combined with manipulative reduction was used to correct fracture displacement preoperatively.All fractures were stabilized by minimally invasive percutaneous plate osteosynthesis (MIPPO) through single or combined medial,anteromedial and anterolateral approaches while minimizing damage to bone attachment and continuity of soft tissue,after soft tissue swelling subsided.For the patients with articular surface collapsing with severe comminution,a series of procedures were done under direct vision including using the talus articular surface as a mold,stable fixation with fine Kirschner (1-1.5 mm) and thin screws (2.1-2.7 mm series) and impaction bone grafting below subchondral bone.Thereafter,distal tibia anatomical short multi-directional locking plate fixation,distal nail support and early ankle joint functional exercise were done.Burwell-Charnley radiological evaluation system was used for radiological assessment,and TeenyWiss scoring system for ankle clinical symptoms and function.Postoperative complications were recorded.Results Follow-up lasted for 11-38 months (mean,16.6 months).No infection,wound disunion,or plate exposure occurred.Burwell-Charnley radiological evaluation system showed anatomic reduction in 32 patients,unsatisfactory reduction in one,and poor reduction in one.According to the Teeny-Wiss scoring system,the results were excellent in 31 patients,good in two and poor in one,with the excellentgood rate of 97％.Three patients suffered traumatic arthritis after operation and alleviated after oral administration of painkiller.Conclusion With use of full reset combined with manipulative reduction to correct fracture displacement,minimally invasive locking plate,distal row of nails,impaction bone grafting and limited fixation,the patients with extremely distal tibial pilon fractures achieve satisfactory reduction,stable fixation,and early functional exercise.

Objective: To observe the effect of the intravenous infusion of vasostatin-2 (VS-2) on hemodynamics in experimental rats with spontaneous hypertension (SH).
Methods: A total of 36 (14-16) weeks male SH rats with the mean body weight at (160-250) g were randomly divided into 6 groups:①Control group, the rats received normal saline (100μl/kg),②Catestatin (20μg/kg) group,③VS-2 (5μg/kg) group,④VS-2 (10μg/kg) group,⑤VS-2 (20μg/kg) group and⑥VS-2 (40μg/kg) group. n=6 in each group. The average blood pressure (BP), heart rate (HR) and barorelfex sensitivity (BRS) were monitored and compared upon VS-2 treatment and between VS-2 and catestatin treatments in conscious and freelance rats.
Results: Compared with prior treatment, VS-2 (20μg/kg) and VS-2 (40μg/kg) could obviously decrease the HR, BP and BRS in SH rats. In VS-2 (20μg/kg) group, HR by bpm was (341.3 ± 19.3) vs (365.5 ± 25.5), BP by mmHg was (133.0 ± 8.9) vs (147.5 ± 11.2) and BRS by ms/mmHg was (0.52 ± 0.18) vs (0.37 ± 0.12);in VS-2 (40μg/kg) group, HR was (348.8 ± 30.8) vs (374.5 ± 34.8), BP was (131.5 ± 9.3) vs (151.7 ± 10.8) and BRS was (0.53 ± 0.05) vs (0.38 ± 0.03), all P<0.01. Catestatin treatment could also decrease the HR as (318.7 ± 13.4) vs (365.5 ± 25.5), BP as (119.7 ± 7.3) vs (147.5 ± 11.2) and BRS as (0.58 ± 0.15) vs (0.35 ± 0.11), all P<0.01. Compared with catestatin (20μg/kg), the rats received VS-2 (20μg/kg) had the weaker reduction of HR as (318.7 ± 13.4) vs (341.3 ± 19.3), BP as (119.7 ± 7.3) vs (133.0 ± 8.9), all P<0.01, while BRS was similar as (0.58 ± 0.15) vs (0.52 ± 0.18), P>0.05.
Conclusion: Intravenous infusion of VS-2 may obviously affect HR, BP and BRS in experimental SH rats;compared with the same dosage of catestatin, VS-2 had the weaker reduction of HR, BP and BRS.