Objective To detect the mutant prevention concentration (MPC) of macrolide resistant Streptococcus pneumoniae, to amplify the resistance gene of Streptococcus pneumoniae mutants selected in the mutant selection window (MSW), and to investigate the mechanism in the resistance of Streptococcus pneumoniae to macrolides. Methods The Streptococcus pneumoniae strain ATCC49619 was enriched in broth, and the bacterial concentrations were adjusted to 1010 colony forming units per milliliter. The minimal inhibitory concentration (MIC), MIC for 99 % of input cells (MIC99), provisional MPC (MPCpr), and MPC of roxithromycin and azithromycin for Streptococcus pneumoniae were determined by agar plate dilution method. The ermB and mefA of Streptococcus pneumoniae mutants selected in the MSW were obtained by PCR method and sequencing. Results The MPCs of roxithromycin and azithromycin for ATCC49619 were 0.80 ?g/ml and 0.51 ?g/ml, and the MPC/MIC99 were 5.0 and 3.9. The ermB gene was detected in Streptococcus pneumoniae mutants selected in the MSW. Conclusion Adjustment of the drug dose may limit the enrichment of macrolide-resistant Streptococcus pneumoniae mutant. The mechanism in the resistance of Streptococcus pneumonia to macrolides may be associated with ermB genes carried.

BACKGROUND:There are numerous studies on bone marrow mesenchymal stem cells(BMSCs)from small animals such as rats and rabbits,but no few reports addressing BMSCs from big animals.OBJECTIVE:To observe in vitro cultured goat BMSCs,and to understand its biological properties.METHODS:A healthy Chinese goat aged ten months was obtained to extract 5 mL fresh bone marrow from the posterior superior iliac spine by puncture following anesthesia.Using the whole bone marrow method,the samples were incubated in a sterile plastic culture flask and added with DMEM/F12 containing 10% fetal bovine serum.Following 80% 90% confluence,cells were digested by trypsin.Cells at passage 3 in logarithmic phase were collected and frozen,and then recovered.Changes in cell morphology were observed using an inverted microscope.Cell growth curve was measured using MTT assay.The potential of osteogenic differentiation was examined utilizing Von Kossa's staining.RESULTS AND CONCLUSION:The primary cultured BMSCs were cultured with adherent growth.The cells were spindle form.Cell morphology following passage 3 was similar,showing long spindle shape.Following freezing and recovery,cell adherence was slower compared with subculture cells,and no significant difference was detected in cell morphology and viability compared with subculture cells.Growth cycle was similar in passage 3-passage 5 cells.BMSCs entered lag phase at days 2 and 3,logarithmic phase at day 3,and platform phase at days 6 and 7,and then growth speed was slow.Goat BMSCs were positive for Von Kossa's stain at 3 weeks following osteogenic induction.Results verified that cultured goat BMSCs showed strong genetic stability and proliferation ability,and differentiated into osteoblasts.

Objective To study the expression of the excision repair cross complementing 1 ( ERCC1 ) and thymidylate synthase ( TS) in gastric cancer and to investigate the correlation between the ERCC 1 and TS ex-pression with the efficacy of FOLFOX adjuvant chemotherapy in gastric cancer patients .Methods A total of 73 gastric cancer patients received modified FOLFOX adjuvant chemotherapy .Real-time PCR was used to investi-gate the expression of ERCC 1 and TS in gastric cancer tissue .We also studied their correlation and association with prognosis .Results ERCC1 and TS expressed in gastric cancer tissues with the positive rate were 42 .5%(31/73)and 61.6%(45/73).There was no significant correlation between the ERCC 1 or TS expression and the clinical features .In this study ,the median of RFS and MST were 16 months and 27 months in patients with ER-CC1-negtive expression,9 months and 16months in patients with ERCC1-positive expression,the median of RFS and MST were significantly different in the two groups (P=0.000,P=0.002).There were no statistical differences on TS expression pattern in either median of RFS or MST (P=0.10,P=0.71).Conclusion The ex-pression of ERCC1 may be useful in prediction of the clinical outcome in gastric cancer patients who treated with FOLFOX7 adjuvant chemotherapy .Patients with ERCC1-negtive expression may benefit from the therapy .

Objective To review our clinical experience with primary surgical repair of tetralogy of Fallot in neonates and premature infants and to discuss the timing of repair and major factors in treating this patients perioperatively.Methods From January 2012 to September 2015,a series of 19 consecutive neonates and premature infants(M/F =12/7) with tetralogy of Fallot were admitted in our center to receive surgical treatment,with a mean age of(17.3 ± 5.5) days(12-28 days) and a mean body weight of(2.9 ±0.7) kg(2.1-4.3 kg).All the 19 cases were symptomatic with cyanosis,saturation on room air 0.79 ± 0.12 (0.48-0.92),and shortness of breath.Before operation,2 cases were receiving an infusion of prostaglandin E1,5 were mechanically ventilated.7 were more than moderate anemia with hemoglobin of 55-87 g/L.All the patients received echocardiography,ECG and chest X-ray.The McGoon ratio and Nakata index were 1.09 ± 0.30 (0.8 to 1.6) and (135.5 ± 54.2) mm2/m2 (63-212 mm2/m2) respectively.18 cases received one-stage surgical repair and 1 premature infant under two-stage operation with the VSD closure after right ventricular outflow tract(RVOT) transannular patch augmentation.All the VSDs in the 18 cases were closed with continuous suture and RVOT were enlarged with autologous pericardium patch transannularly or not.Balanced and modified ultrafiltration were applied in all the patients.Patients were strictly followed up with a standard protocol focusing on right ventricular function and arrythmia.Several characteristics(e.g.time of operating,mechanical ventilation and ICU stay,complications,hospital stay time,cost of hospitalization) were compared between this group of patients and other TOF patients during the same period in our center.Results All the one-stage operations were successful.There was no mortality and major complication.Mean CPB and aortic clamping time were(111.5 ± 31.6)min (76-153min) and (73.3 ± 11.6) min (64-89 min) respectively.10 VSDs were closed with transventricular approach,6 witht transatrial appraoch and 2 with transatrial-ventricular approach.12 cases (66.7 ％) had a transannular RVOT patch,4 (22.2 ％) with single RVOT patch and 2 (11.1％) with transannular RVOT and left pulmonary artery patch.Atrial communication were left open in 15 cases (83.3 ％).The time of mechincal ventilation and ICU stay were (123.7 ± 59.5) h (39-239 h) and (10.1 ± 3.2) days (5-19 days) respectively.All the patients were followed up for (31.8 ± 15.7) months (9-57 months).There was no mortality and major complication.ECG showed that there was no severe arrythmia except for 3 complete right branh bundle block.The latest echocardiography results showed that right heart function was normal in all the cases and RVOT grandients was less than 30mmHg except one with 35 mmHg.There were 2 cases with residual shunt less than 2 mm and 8 cases with pulmonary valvular regurgitation less than moderate degree.Compared with other TOF cases during the same period,there was no difference according to the data mentioned above except with more time of mechnical ventilation and ICU stay and more cost of hospitalization.Conclusion Primary repair of TOF can be performed safely in symptomatic neonates and premature infants,regardless of age and body weight,with favorable early and mid-term results.Excellent teamwork and accurate prenatal and postnatal diagnosis were the two major factors in yeilding good results in these patients.

Iron uptake mechanism of Vibrio alginolyticus was primarily investigated. V.alginolyticus could survive in the medium with high-concentration iron chelator. The strain of V. alginolyticus isolated from diseased fish produced more siderophore than that from marine environment. The extract of siderophore from V. alginolyticus could stimulate the growth of Escherichia coli mutant AN93. Under iron limitation,the growth rate was decreased and several outer membrane proteins were induced. Adding iron into the iron-limited medium the normal growth could be recovered.

Background Choroidal thickness is closely associated with ametropia,and to study the influence of subfoveal choroidal thickness (SFCT) on refraction is very important for understanding the mechanism of refractive error more clearly.Objective This study was to investigate the relationship between SFCT and refraction.Methods A retrospective serial cases analysis was performed.Forty anisometrope patients were recruited in Zhongshan Ophthalmic Center from June 2012 to August 2012.The subjects were divided into 6-13 years group and 14-21 years group.Vision acuity was tested by the EDTRS visual acuity chart and ocular anterior segment was examined under the slit lamp microscope,and the fundus examination was performed by direct ophthalmoscope.Subjective and objective optometry was performed after pupils were dilated.EDI OCT was used to illuminate choroidal image.Ocular axial length (AL) was obtained by Lenstar 900.The difference in SFCT between myopic eyes and hyperopic eyes was compared using Student t test,and the correlations between SFCT and refraction were analyzed by Pearson linear analysis and linear regression analysis.Results The average SFCT was (307.82±88.47) μm in all the tested eyes,and the SFCT of myopic eyes and hyperopic eyes was (270.60±70.57) μm and (376.95±76.59) μm,respectively,with a significant difference between them (P =0.000).In the 6-13 years group,positive correlations were found between SFCT and diopters with the regression coefficient 18.60 and regression equation Y =18.60X +310.79 (r=0.345,F=21.110,P=0.000) as well as between SFCT and AL with the regression coefficient -31.76 and regression equation Y =-31.76X+1 039.97 (r=0.262,F=17.320,P=0.000).In the 14-21 years group,SFCT showed positive correlation with diopters,with the regression coefficient 23.38 and regression equation Y=23.38X+353.17 (r =0.430,F =27.210,P =0.000) and negative correlation with AL,with the regression coefficient-35.82 and regression equation Y =-35.82X+1 132.75 (r=0.237,F=15.650,P=0.000).Conclusions SFCT seems to be positive correlated with diopter.When the diopter shifts toward positive value,SFCT value increases,and whenever diopter increases-1 D,SFCT decreases 20 μm approximately.SFCT decreases with the increase of AL.

Objective To evaluate the clinical effect of long incision and reticular incision in the treatment of osteofascial compartment syndrome(OCS)in children.Methods The study involved 56 children with OCS who met failure of the conservative treatment.The injury sites included mainly the forearm and the leg.All the children underwent decompression by a small reticular incision procedure from January 2000 to May 2009.The fractures were treated with one stage reduction and fixation or second stage open reduction.Meanwhile,the study involved another 21 children(including 13 earthquake victims)who were treated with long incision for open decompression in the other hospitals before admission.All the wounds were healed by direct suture or dermatoplasty after 1-5 weeks of infection control.Bacterial culture was performed in all the wounds.The fractures were treated with secondary open reduction and fixation.Bacteria culture was done in all wounds.Results The reticular incisional wounds of 56 patients were healed free from dermatoplasty,with no infection or sensory dysfunction.Among 21 patients treated with long incision,the bacteria culture was positive in 16 patients(including 13 earthquake victims)and verve injury found in five patients(including two with radial nerve injury and three with peroneal nerve injury).Four patients with partial or complete cut-off of the verves were improved after repair by secondary operation.Direct suture of incisions was done in nine patients and skin graft performed in 12,with average healing time for three weeks.The patients were followed up for mean 5.6 years,which showed no claw hands,with overall satisfactory rate for about 98% and 95% respectively in two groups according to the probation standard of amputated finger function evaluation from the Hand Surgery Plant of Chinese Medical Association.Conclusions Both long incision and reticular incision procedures can be used early in the treatment of pediatric OCS,while the reticular incision procedure is more convenient and simple,with less complications.

Objective To determine the effects of histone deacetylase inhibitor suberoylanilide hydroxamic acid(SAHA) on the cell proliferation and apoptosis of the human hepatic stellate cell line LX-2.The possible underlying mechanisms were also investigated.Methods The LX-2 cells were treated with SAHA in vitro.The morphology of LX-2 cells in different concentrations groups was observed by inverted microscope;the proliferation of LX-2 cells was measured by MTT assay;the Annexin V-FITC and PI staining was used to detect the apoptosis of LX-2 cells by flow cytometry and fluorescence microscope;the expression of α-SMA,collagen Ⅰ,acH3K9,acH3K14 and acH3K18 were detected by Western blot.Results The morphology change of LX-2 cells showed that SAHA inhibited the proliferation rate of LX-2 cells and in a dose dependent manner(P<0.05).The LX-2 cells were sensitive to SAHA along with time increasing,and in a time-dependent manner(P<0.05).Western blot showed that the expression levels of α-SMA and collagen-Ⅰ were significantly lower(P<0.05),on the contrary,the acetylation levels of acH3K9,acH3K14 and acH3K18 were significantly higher (P<0.05).Conclusions The increased acetylation of the histone acH3K9,acH3K14,acH3K18 and the lower expressed α-SMA and collagen-Ⅰ in LX-2 cells may be one of the mechanisms of SAHA.

Objective To investigate the effect of intramedullary stabilization of lower limb fractures with titanium elastic nails (TENs) in children.Methods From June 2004 to October 2010,278 children were stabilized with TENs in our hospital,including 181 boys and 97 girlsat average age of 7.6 years (range,3.0-14 years).There were 162 patients with femoral fractures,of which 35 underwent open reduction and the rest closed reduction.There were 116 patients with tibial fractures,which was treated with closed reduction.Postoperative fixation lasted for 3-4 weeks.The mean hospital stay was seven days.Of all the patients,272 patients received the removal of the TENs six months after operation and 242 received more than one year of follow-up to have a further observation on the limb length,hip and knee mobility and gait condition in children.Results All the patients were followed up for 6-66 months (mean,33 months).All fractures were healed in good alignment without severe complications.There were no intraoperative complications like infections,delayed union,nonunion,TEN broken and femoral or tibial epiphysis injuries.Ten patients suffered from swell and skin irritation in the nail' s position 3-4 months after operation,and the nails were removed in advance.Nineteen patients showed unequal lengths ( within 1 cm) of the lower limbs,with normal function of the lower limb joint but with no obvious limp.According to the Flynn evaluation standard,the excellent and good rate was 100％.Conclusions Characterized by good stability,rapid healing,small incision,anatomic reduction,unimpaired periosteum,short hospital stay and few complications,intramedullary stabilization with titanium elastic nails can achieve effective fixation in the treatment of lower limb fractures in children.

ObjectiveTo investigate the clinical effects of minimally invasive incision in anterior traverse approach for Gartland type Ⅲ humeral supracondylar fractures in children.Methods Forty-two patients with Gartland type Ⅲ displaced supracondylar fractures of the humerus were treated through anterior traverse approach between January 2008 and April 2011.The fractures were fixed using two Kirschner wires from the medial and lateral epicondyles placed crossing each other.There were 31 males and 11 females,at age of 2.6-12 years (average 5.7 years).Three patients were with open fractures.Seven patients were combined with other fractures,four with radial nerve injury and one with median nerve injury,with time from injury to hospitalization for a range of 1 hour to 3 days.Of all the patients,30 patients were with fractures on the right side and 12 with fractures on the left side.All fractures were treated by minimally invasive incision in anterior traverse approach,when the neurovascular and muscular probing and repair were performed. Results Forty patients were followed up for 3-32 months ( average 13 months),which showed incision healing at one stage,with fracture healing time for 3-4 weeks (average 3.6 weeks).No cubitus varus or incision scar were found after operation.According to Flynn' s criteria,the effect of treatment three months after operation were excellent in 32 patients,good in four and fair in four,with excellence rate of 90％.ConclusionsThe minimally invasive incision of anterior traverse approach is characterized by minor trauma,small incision,nil scar and convenient neurovascular and muscular exploration and hence is a safe and reliable treatment for open reduction of Gartland type Ⅲ humeral supracondylar fractures in children.