Objective To develop an assay to determine Mycobacterium tuberculosis resistance to rifampin, isoniazid, ofloxacin and amikacin by pyrosequencing and evaluate the value of this method in clinical application. Methods Eighty-nine clinical isolates of Mycobacterium tuberculosis from tuberculosis patients were collected from Shanghai Pulmonary Hospital during 2008 to 2009. All strains were isolated from decontaminated sputum, cultured on Lowenstein-Jensen media and identified by traditional biochemical tests with standard methods. Ten Mycobacterium tuberculosis were selected from the strain bank of Shanghai Pulmonary Hospital. The optimal conditions of detection of rpoB, katG, gyrA and rrs gene by pyroseuencing were determined, using the 10 Mycobacterium tuberculosis strains whose drug susceptibility of Bactec 960 and sequence of rpoB, katG, gyrA, rrs gene were known. Then the drug susceptibility of 89 Mycobacterium tuberculosis clinical isolate strains were detected by pyrosequencing using this conditions and the results were compared with that of the Bactec 960 methods. Results The pyrosequencing program of sequence analysis was suitable for the detection of the mutations of rpoB and gyrA genes, and the program of single nucleotide polymorphism was suitable for katG and rrs genes. Among the 89 Mycobacterium tuberculosis clinical isolates,when using the drug susceptibility of Bactec 960 as the standard, the sensitivity of rifampin, isoniazid,ofloxacin and amikacin is 98.0%, 64. 1%, 79.5%, 78. 3% respectively, the specificity is 97.5%,100. 0%, 90. 0%, 100. 0% respectively, the accuracy is 97.8%, 77. 5%, 85.4%, 94. 4% respectively,tested by pyrosequencing. The results of the detection of resistance to rifampin, isoniazid, ofloxacin and amikacin in Mycobacterium tuberculosis using pyrosequencing technique were almost the same with that of Bactec 960, and Kappa ≥0. 7 in each detection. Conclusion Pyrosequencing is thus a rapid, accurate and high throughput method for detecting Mycobacterium tuberculosis resistance to these four drugs.

Objective:To transfect the non-viral vector polyethylenimine (PEI)mediated miR-2861 mimic into the MC3T3-E1 cell line,and to explore the transfection efficiency of PEI/miR-2861 complex and its effects on the proliferation and osteogenesis differentiation in pre-osteoblasts. Methods:The proper amount of PEI was blended with miR-2861 mimic and negative control (NC)separately in a ratio of N∶P=10∶1,and they were divided into experiment group and NC group. The NC/PEI complex acted as the NC group was used to eliminate the interference of osteogenesis from the addition of double-stranded RNA mimic.MTT assay was used to determine the optimal concentration of PEI/miR-2861 mimic complex.The fluorescence imaging technique and bulge-loop RT-PCR were used to detect the transfection efficiency and mRNA expression of miRNA-2861 in the cells with different concentrations (10,30, 50,and 100 nmol · L-1 ), separately.The osteogenesis ability of MC3T3 cells was identified with RT-PCR and Alizarin red staining with the selected concentration of PEI/miR-2861 by transient transfection.Results:Compared with blank control group,the proliferation rates of MC3T3 cells in 100 nmol·L-1 PEI/miR-2861 group was decreased significantly at 72 h (P < 0. 05 ). With the increasing of transfected concentration the transfection efficiency of miRNA/PEI complex was increased gradually.The results of Alizarin red staining and quantitative analysis showed that calcium deposits were more and bigger in experiment group after induced for 21 d,while both in blank control group and NC group they were less.Conclusion:The miRNA-2861 mimic can be effectively transfected into the MC3T3-E1 cell line and expresses with a high level,which is mediated by PEI as the gene vector.miR-2861 mimic has a certain ability of promoting osteogenesis differentiation of MC3T3-E1 cells.

Objective:To study the bone-conductivity and absorbability of nano-hydroxyapatite/collagen(nHAC) composite implanted under the calvarial periosteum in rabbits.Methods:24 nHAC samples and 24 HA samples were prepaired in the shape of round disk with the diameter of 8 mm and thickness of 3 mm.nHAC samples were implanted under calvarial periosteum on the left side and HA samples on the right of 24 rabbits.The bone-conductivity and absorbability of the samples were examined by new bone height measuring and residual implant materia measuring 2,4,8 and 12 weeks after operation. Results:2,4,8 and 12 weeks after implantation new bone height(mm) in nHAC group was 0.54?0.09,0.72?0.12,1.83?0.14 and 2.63?0.07,that in HA group 0.13?0.11,0.31?0.12,1.23?0.05 and 1.75?0.14,respectively(P

Objective To find out the resistant situation and drug of Mycobacteria patients in Sichuan and offer foundation for clinical.Methods Two hundred randomized clinical isolates of Mycobacterium were determined by Roche drug sensitivity and minimum inhibitory concentration (MIC) method.Results Of the 200 clinical isolates,192 stains were Mycobacterium tuberculosis(MTB) (96.0％),8 strains (4.0％) were non-tuberculosis mycobacterium(NTM).Of the 192 MTB strains,108( 57.3％ ) sensitive strains and 84 (43.7％)stains were resistant to one or more than one drugs.Among these 84 resistant strains 23 were multi-drug resistant ( MDR,12.0％ ),4 were extensively drug resistant( XDR,2.1％ ).The anti-TB drug resistance rates were:SM(16.7％),INH(20.8％),RFP(17.2％),EMB(10.9％),PI(16.1％),LFX(8.8％),AMK ( 16.7％ ),CPM ( 6.2％ ),PTA ( 33.3％ ),respectively.Conclusion The resistance rate of tuberculosis keeps at a high level in Sichuan,especially the resistance rate of multiple (≥4) drug,we should oar attention.

Objective: To investigate the value of shear wave elastography(SWE) to evaluate local advanced rectal cancer after neoadjuvant radiochemotherapy. Methods: In a retrospective study, endorectal ultrasound(ERUS) and endorectal SWE were performed in 73 patients with local advanced rectal cancer before and after neoadjuvant radiochemotherapy. The mean and maximum values of Young′s modulus for SWE to evaluate the lesions before and after neoadjuvant radiochemotherapy were recorded. According to the postoperative pathological T stage, the lesions were divided into reduction of T stage group and non-reduction of T stage group. The efficacy of ERUS in diagnosing reduction of T stage was calculated, and the differences of the mean and maximum values of Young′s modulus between reduction of T stage group and non-reduction of T stage group was calculated, and the differences between the two groups were compared. ROC curves were constructed by the difference of mean and maximum Young′s modulus of lesions before and after neoadjuvant radiochemotherapy, respectively, to evaluate the diagnostic value of the difference in predicting reduction of T stage. Results: A total of 57 cases had reduction of T stage after neoadjuvant radiochemotherapy (57/73, 78.1%). The mean and maximum values of Young′s modulus before and after neoadjuvant radiochemotherapy were compared, and the differences were statistically significant(all P<0.01). After neoadjuvant radiochemotherapy, the values of Young′s modulus of the lesions increased with the increase of pT stage. Compared with the mean values of Young′s modulus of the lesions in pT3 stage, the differences of the mean values of Young′s modulus of the lesions in pT0, pT1 and pT2 stages were statistically significant(all P<0.01). Compared with the maximum values of Young′s modulus of the lesions in pT3 stage, the differences of the maximum values of Young′s modulus of the lesions in pT0 and pT1 stage were statistically significant(all P<0.01). The differences of the mean value and the maximum value of Young′s modulus in the reduction of T stage group and the non-reduction of T stage group was statistically significant(all P<0.01). The ROC curve was established and determined by calculation. Taking the average difference of 34.7 kPa as the best diagnostic threshold, the average hardness of the lesion after neoadjuvant radiochemotherapy decreased more than 34.7 kPa to diagnose the reduction of T stage, the sensitivity, specificity and accuracy were 87.7%, 93.8% and 89.0%, respectively. Compared with ERUS, the difference was statistically significant(P=0.032). Conclusions Shear wave elastography is an effective technology to help ERUS in evaluating the lesions of rectal cancer after neoadjuvant radiochemotherapy and has a promising future.

Wandaitang, which is one of classical traditional Chinese medicine(TCM) prescriptions, is derived from Collected Exegesis of Recipes of Fu Qingzhu' s Obstetrics and Gynecology. It is commonly used in modern clinical practice, and included in the Catalogue of Ancient Classical Prescriptions (The First Batch). Collected Exegesis of Recipes of Fu Qingzhu' s Obstetrics and Gynecology and Chen Shi-duo' s Bianzhenglu have a complicated relationship. Both of them have another biography, named Nvke Xianfang and Bianzheng Qiwen. The composition of Wandaitang in the four books is slightly different, while the prescription's explanations and other records are almost the same. The research and development of Wandaitang should be based on the records of Collected Exegesis of Recipes of Fu Qingzhu's Obstetrics and Gynecology. Compared with other classical literatures, Collected Exegesis of Recipes of Fu Qingzhu's Obstetrics and Gynecology was published in a late period and less reproduced in other ancient books. To study the function of Wandaitang, we need to analyze the records in the original book. In addition, we need to make a multi-angle analysis by reference to the theory of TCM, the composition of drugs, the significance of compatibility, as well as the understanding of modern famous doctors, clinical reports and experimental studies in all aspects. The study found that the functions of Wandaitang were relatively concentrated, but with wide major functions involving internal medicine, surgery, gynecological, pediatric, andrological and other departments. According to the study, the authors believe that the functions of the classical TCM prescription of Wandaitang are invigorating spleen to eliminate dampness, dispersing the liver and rectifying Qi, and invigorating Yang. It can be used to treat leucorrhea, diarrhea, edema and stranguria with the syndromes of pale, languid, little food, loose stool and depression. Wandaitang can also be used to treat vaginitis, cervicitis, pelvic inflammatory disease, irritable bowel syndrome, chronic colitis, chronic nephritis, nephrotic syndrome and chronic prostatitis.

OBJECTIVETo evaluate the therapeutic effect of integrative Chinese and Western medicine in treating mid-advanced crescentic nephritis (MACN).

METHODSThirty-two patients, their diagnosis was confirmed as MACN by renal biopsy, were divided, adopting randomized, controlled method, into two groups, the treated group and the control group, they were all, excepting one, treated with impact therapy of methyl-prednisolone followed with oral intake of prednisone, to part of them cyclophosphamide or mycophenolate mofetil was given in addition, to those with hypo-hemoglobin (< 90 g/L), subcutaneous injection of erythropoietin was administered. Decoction of Qingre Huoxue recipe (QHR), consisted of oldenlandia herb 30 g, honey-suckle stem 30 g, violet herb 30 g, red peony root 15 g, rehmannia root 15 g, solomonseal rhizome 15 g, asiabell root 30 g, red sage root 30 g, prepared rhubarb 12 g and giant-hyssop herb 12 g, were additionally given one dose per day to patients in the treated group. The renal function, improvement of anemia and immunosuppressive agents needed in patients were observed after 3 months treatment.

RESULTSAfter treatment, renal function was improved in both groups, but the effect in the treated group was better than that in the control group (P < 0.05). Anemia was partially alleviated in the two groups with no significant difference. The dosage of glucocorticoids used in the treated group was obviously lesser than that used in the control group (P < 0.01).

CONCLUSIONIntegrative Chinese and western medicine could treat crescentic nephritis to obtain good effect, and reduce the quantity of glucocorticoid necessity for treatment.

Adult ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glomerulonephritis ; drug therapy ; physiopathology ; Humans ; Immunosuppressive Agents ; therapeutic use ; Kidney Function Tests ; Kidney Glomerulus ; physiopathology ; Kidney Tubules ; physiopathology ; Male ; Methylprednisolone ; therapeutic use ; Middle Aged ; Phytotherapy

Objective To explore the effect of early administration of high-dose amino acids in parenteral nutrition for low birth weight infants of prematurity.Methods A total of 191 prematures admitted to the First Affiliated Hospital of Nanjing Medical University from June 2015 to December 2016 were selected,and they were randomly divided into the study group (n =110) and the control group (n =81).The starting amino acid dose was 1.8-2.5 g/(kg · d)and 1.0-1.5 g/(kg · d) in the study group and the control group,respectively.Lipids,glucose,and electrolytes in parenteral nutrition were applied according to standard protocol of the guideline.And the start of enteral feeding,the recovery of birth weight,the duration of parenteral nutrition,the time before total enteral nutrition,duration of hospital stay,incidence of respiratory distress syndrome,duration of mechanical ventilation and incidence of kaliopenia were compared between the 2 groups.Results The start of enteral feeding,the recovery of birth weight,the duration of parenteral nutrition,the time before total enteral nutrition in the study group were earlier than those in the control group [(3.83 ±3.15) d vs.(5.53 ±5.63) d,(15.47±10.54) d vs.(19.47 ± 14.57) d,(16.46 ± 10.33) d vs.(21.41 ±18.00) d,(6.36 ± 4.88) d vs.(8.48 ± 9.27) d],and the differences were all statistically significant (t =2.455,2.097,2.217,2.041,P =0.016,0.038,0.029,0.043).The duration of hospital stay was shorter and the hospitalization expenses were lower in the study group than those in the control group,but the differences were not significant (all P ＞ 0.05).The incidence of respiratory distress syndrome in the study group was significantly lower than that in the control group[20.91％ (23/110 cases) vs.35.80％ (29/81 cases)],and the difference was statistically significant (x2 =5.223,P =0.022).The duration of mechanical ventilation in study group was shorter than that in control [(1.12 ± 2.62) d vs.(3.31 ± 8.13) d],and the difference was statistically significant (t =2.231,P =0.028).The incidence of kaliopenia in the study group was higher than that in the control group [30.91％ (34/110 cases)vs.17.28％ (14/81 cases)],and the difference was statistically significant (x2 =4.603,P =0.032).There were not significant differences in terms of complications of necrotizing enterocolitis,sepsis,extrauterine growth retardation,hospital infection,metabolic acidosis,hyperglycemia,glucopenia,patent ductus arteriosus and respiratory distress syndrome between 2 groups (all P ＞ 0.05).Conclusions Early administration of high-dose amino acids in parenteral nutrition for preterm infants can result in earlier enteral nutrition,shorter parenteral nutrition duration without increasing incidence of complications and hospitalization expenses.

Objective To evaluate the value of endorectal elastography with strain ratio to estimate local advanced rectal cancer after neoadjuvant radiochemotherapy.Methods In a retrospective study, endorectal ultrasound,endorectal elastography and enhanced rectal MRI were performed in 67 patients with local advanced rectal cancer after neoadjuvant radiochemotherapy.The imaging results were compared with postoperative pathological T stage and NCCN TRG.Results There was no significant difference in the diagnosis accuracy between T stage of ERUS(55.2%)and MRI(56.7%).Endorectal elastography results showed lesions confined to the rectal wall(T0-2 stage)were softer than lesions invaded the peripheral fat (T3)and the difference was statistically significant(P <0.05).When the cut-off point was set at SR<2.78,the sensitivity,specificity and accuracy of diagnosis of T0-2 were 64.7%,87.5% and 70.1% respectively.The lesion tended to have a greater SR value when residual tumor components increased(a higher NCCN TRG).Conclusions Endorectal elastography is an useful and effective imaging method to evaluate local advanced rectal cancer after neoadj uvant radiochemotherapy.It can help ERUS and rectal MRI to evaluate the lesions.

Objective To investigate the value of shear wave elastography ( SWE) to evaluate local advanced rectal cancer after neoadjuvant radiochemotherapy . Methods In a retrospective study ,endorectal ultrasound( ERUS) and endorectal SWE were performed in 73 patients with local advanced rectal cancer before and after neoadjuvant radiochemotherapy . T he mean and maximum values of Young′s modulus for SWE to evaluate the lesions before and after neoadjuvant radiochemotherapy were recorded . According to the postoperative pathological T stage ,the lesions were divided into reduction of T stage group and non‐reduction of T stage group . T he efficacy of ERUS in diagnosing reduction of T stage was calculated ,and the differences of the mean and maximum values of Young′s modulus between reduction of T stage group and non‐reduction of T stage group was calculated ,and the differences between the two groups were compared . ROC curves were constructed by the difference of mean and maximum Young′s modulus of lesions before and after neoadjuvant radiochemotherapy ,respectively ,to evaluate the diagnostic value of the difference in predicting reduction of T stage . Results A total of 57 cases had reduction of T stage after neoadjuvant radiochemotherapy ( 57/73 ,78 .1% ) . The mean and maximum values of Young′s modulus before and after neoadjuvant radiochemotherapy were compared ,and the differences were statistically significant ( all P ＜ 0 .01) . After neoadjuvant radiochemotherapy ,the values of Young′s modulus of the lesions increased with the increase of pT stage . Compared with the mean values of Young′s modulus of the lesions in pT3 stage , the differences of the mean values of Young′s modulus of the lesions in pT 0 ,pT1 and pT2 stages were statistically significant ( all P ＜ 0 .01 ) . Compared with the maximum values of Young′s modulus of the lesions in pT3 stage ,the differences of the maximum values of Young′s modulus of the lesions in pT0 and pT1 stage were statistically significant( all P ＜0 .01) . T he differences of the mean value and the maximum value of Young′s modulus in the reduction of T stage group and the non‐reduction of T stage group was statistically significant ( all P ＜ 0 .01 ) . T he ROC curve was established and determined by calculation . Taking the average difference of 34 .7 kPa as the best diagnostic threshold ,the average hardness of the lesion after neoadjuvant radiochemotherapy decreased more than 34 .7 kPa to diagnose the reduction of T stage ,the sensitivity ,specificity and accuracy were 87 .7% ,93 .8% and 89 .0% ,respectively . Compared with ERUS , the difference was statistically significant ( P ＝ 0 .032 ) . Conclusions Shear wave elastography is an effective technology to help ERUS in evaluating the lesions of rectal cancer after neoadjuvant radiochemotherapy and has a promising future .