Aim To establish a resistant human osteosarcoma cell line(Saos-2/ADM1and Saos-2/ADM4)from the Saos-2 cell line and study its resistant mechanisms.Methods Saos-2 cells were pulse exposed in gradually increased dose of ADM culturemedium.The sensitivity of the resistance drug from the Saos-2、Saos-2/ADM1 and Saos-2/ADM4 cell lines to ADM、DDP、EPI、MTX、THP and PTX was measured by MTT assay.The morphology and ultramicro structure of the cell lines were observed by optical microscopy and transmission electronic microscopy.The expression level of MDR1 mRNA、MRP mRNA and their proteins P-gp、MRP was examined by reverse transcription polymerase chain reaction and immunohistochemistry.Results Resistance cell lines were established after 167 days.The resistance index to methotrexate of the Saos-2/ADM1and Saos-2/ADM4 cell lines to ADM was 49.8 and 74.6 times than that of Saos-2 respectively.The two cell lines had resistance to MTX、IFO、EPI、THP and PTX(P0.05).Disordered structure of the Saos-2/ADM1and Saos-2/ADM4 cells was observed through microscopy.The cells appeared in coenocytic.The decrease of cell villus and the increase of nucleoli were observed through transmission electronic microscopy.The proliferation ability of Saos-2/ADM1and Saos-2/ADM4 cells decreased significantly.MDR1 mRNA、MRP mRNA、P-gp and MRP showed positive staining in resistance cell lines.Conclusion The genes of MDR1 mRNA、MRP mRNA and their corresponding proteins participated in the formation of multidrug resistance in resistant adriamycin cell line.These newly-described resistant osteosarcoma cell lines were useful models for further characterization of drug resistance in osteosarcoma and for the development of treatment protocol.

Objective:To explore the clinical features and risk factors of systemic inflammatory responses syndrome (SIRS) after flexible ureteroscopic lithotripsy (FURL) with upper urinary tract stones.Methods:A retrospective of 157 patients with upper urinary calculi treated by FURL who were admitted to Suzhou First People′s Hospital from January 2018 to October 2020 were selected to analyze the treatment results and complications of the patients. Patients were divided into SIRS group ( n=31) and non-SIRS group ( n=126) according to whether they were complicated by SIRS. The clinical data between the two groups were analyzed by a single factor, and the factors with statistical significance were included in the LASSO-Logistic regression analysis. Visualize the model through the Nomogram, and verify and evaluate the discrimination and accuracy of the model. Results:The results of univariate analysis showed that gender( P=0.010), average stone size ( P<0.05), preoperative urine white blood cell count ( P<0.05), operation time ( P<0.05) and postoperative stone bacterial culture ( P<0.05) were different between the two groups and the differences were statistically significant. The LASSO-Logistic regression analysis results show that the above factors were all independent risk factors for patients with SIRS. The Nomogram complicated SIRS risk prediction model constructed based on the above risk factors has good discrimination and high consistency with actual observations. Conclusion:Females, large stones, high preoperative white blood cell counts, longer operation time, and positive bacterial culture of postoperative stones are independent risk factors for SIRS after FURL in patients with upper urinary calculi.

Objective To validate the performance of a PMA-based assay for detection of INH-resistant mutations in MTB and investigate the mutation characteristic of INH-resistance.Methods The MTB standard strain H37Rv was from National Tuberculosis Reference Laboratory,1 wild-type strain and 1 katG S315T ACC mutant strain were from Xiamen CDC,7 MTB INH-resistant strains with known INH resistance mutations were from Shenzhen Center for Chronic Disease Control,Henan CDC,No.309 Hospital of PIA and Xiamen CDC.707 MTB clinical isolates were from Xiamen CDC,Xiamen No.1 Hospital and Zhangzhou CDC,126 sputum samples were from Xiamen Tongan CDC.The genomic DNA of the MTB standard strain H37Rv,7 MTB INH-resistant strains and 833 clinical samples,were extracted with Xiamen Zeesan Mycobacterium tuberculosis Isoniazid-resistance Mutation Test Kit using the thermal lysis method.The genomic DNA of 1 wild-type strain and 1 katG S315T ACC mutant strain were extracted with AxyPrep Bacterial Genomic DNA Miniprep Kit.The mutations were discriminated by the △Tm between the samples and wild type control in katG315 codon,inhA promoter -17 to -8 region,ahpC promoter region -44 to -30 and - 15 to 3,and inhA94 codon.A 10-fold dilution series of MTB DNA from 3 × 105 to 300 copies/ reaction obtained from a wild-type strain and a katG S315T ACC mutant strain,respectively,were prepared to determine the analytical sensitivity.Seven MTB INH-resistant strains with 9 predetermined mutations were used for the analytical specificity assay,and 5 mutants of which were used for the repeatability assay.The clinical detection performance of PMA assay were confirmed by the sequencing method in 833 samples.Results Results could be obtained within 3 hours from DNA extraction to PMA assay,including 46 samples in a standard 96-well real-time PCR instrument simultaneously.The analytical sensitivities of PMA were 300 copies/reaction for both the wild-type strain and katG S315T ACC mutant strain.Nine INH-resistant point mutations could be discriminated and 5 of which had standard deviations of melting temperature less than 0.5 ℃.Fully concordant results of mutant locus between PMA assay and sequencing were obtained in all 162 mutant samples.INH-resistant mutations in the four loci were found in 19.4％ (162/833) samples by PMA assay in Xiamen and Zhangzhou.Among the 14 lNH-resistant mutant types detected,katG S315T ( AGC→ACC),inhA promoter - 15C→T and katG S315N (AGC→AAC) accounted for 83.3％ (135/162) of the overall mutations.

Objective To summarize the features of computed tomography (CT) and magnetic resonance imaging (MRi) of hepatic cystic echinococcosis, and investigate the key points of identification and diagnosis.Methods The clinical data of 58 patients with hepatic cystic echinococcosis who were admitted to the Affiliated Hospital of Inner Mongolia Medical University from August 2011 to August 2014 were retrospectively analyzed.Patients received plain and enhanced scan of CT and MRI.Hepatic cystic echinococcosis was divided into the 5 types according to the literatures, including unilocular echinococcasis in type Ⅰ, multivesicular hydatid cysts in type Ⅱ, anechoic content with detachment of laminated membrane from the cyst wall in type Ⅲ, calcification of lesions in type Ⅳ and mixed echinococcosis in type Ⅴ.Patients who were diagnosed as with definite or suspected hepatic cystic echinococcosis underwent surgery.The follow-up including observing the recurrence of hepatic cystic echinococcosis was performed by outpatient examination and telephone interview at postoperative month 3, 6, 12 for 1 year and then once every year up to August 2015, and was ended if there was no recurrence for more than 5 years.Results (1) The results of CT and MRI examinations: of the 58 patients, 54 received scan of CT and 21 received scan of MRI.Seventeen patients were detected in type Ⅰ with clear-boundary and low-density cystic lesions by CT examination;MRI examinations showed there were single or multiple, round or oval abnormal signal including low T1WI signal, high T2 WI signal and low T1 WI and T2WI signal of cyst wall.Thirteen patients were detected in type Ⅱ, CT examination showed the daughter cysts of multiple sizes were found in the mother cyst, arranged in honeycomb or wheel shape;MRI examination showed there were lower T1 WI signal in the daughter cyst and higher T2 WI signal in the daughter cyst compared with signal in the mother cyst, and low signal in the cyst wall of the daughter cyst and mother cyst.Six patients were detected in type Ⅲ with capsule in capsule sign and water snake sign by CT examination and ribbon sign by MRI examination.Thirteen patients were detected in type Ⅳ, CT examination showed there were irregular high-density calcified shadow with the performances for return sample or sample volume skins changes.Nine patients in type Ⅴ had more than 2 kinds of lesions.(2) Diagnosis: 4 patients were misdiagnosed by CT examination including 3 with preoperative diagnosis of hepatic cyst and 1 with preoperative diagnosis of metastatic carcinoma of liver, with an accurate rate of diagnosis of 92.6％ (50/54).Two patients with preoperative diagnosis of hepatic cystic adenocarcinoma were misdiagnosed by MRI examination, with an accurate rate of diagnosis of 90.5％ (19/21).(3) Treatment and follow-up: 58 patients underwent surgery, including 40 undergoing internal capsule removal with external capsule suturing (31 with open operation and 9 with laparoscopic operation), 10 undergoing partial hepatectomy and 8 undergoing external capsule enucleation.Of 58 patients, 3 were complicated with effusion of residual cavity, 2 with unclosed external capsule, 1 with bile leakage and then was cured after 4-8 week drainage.Fifty patients were followed up for 12.0-48.0 months with a median time of 27.1 months and a follow-up rate of 86.2％ (50/58).During the follow-up, 1 patient undergoing internal capsule removal had recurrence at postoperative month 8 and was cured by CT-guided interventional therapy using absolute alcohol, and other patients had no recurrence.Conclusions There was a higher accuracy in CT and MRI examinations for hepatic cystic echinococcosis.Honeycomb and wheel shapes are characteristic findings of hepatic cystic echinococcosis in type Ⅱ.The characteristic performances of CT examination for hepatic cystic echinococcosis in type Ⅲ are capsule in capsuleand water snake signs, and characteristic performances of MRI examination is ribbon sign.The ring-like enhancement of edge by MRI examination is an essential of identification and diagnosis between hepatic cystic echinococcosis and hepatic cyst, and irregular calcification is a differential point between hepatic echinococcosis and hepatic tumor.

Objective To observe the clinical efficacy of the Mingmen eight array massage combined with compound lysine hydrochloride and zinc gluconate granules for the treatment of infantile diarrhea. Methods Totally 120 children with diarrhea met the inclusion criteria were divided into the treatment group, Western medicine group, and general massage group by random number table method, 40 cases in each group. All 3 groups were given dietary guidance and did moderate exercise. Western medicine group was administered orally for compound lysine hydrochloride and zinc gluconate granules, two bags per day for the aged 1–10, three bags per day for the aged >10. Treatment group was given the Mingmen eight array massage therapy based on Western medicine. General massage group received conventional massage therapy based on Western medicine, with four weeks as treatment course. 3 groups of patients were observed before and after treatment for salivary amylase, blood zinc value, urine D-xylose absorption and excretion test, TCM symptom score, and clinical efficacy were evaluated. Results There was statistical significance in the salivary amylase, blood zinc value, and urinary D-xylose absorption and excretion rate between before and after t reatment(P<0.05); after treatment, treatment group showed higher salivary amylase, blood zinc value, and urinary D- xylose absorption and excretion rate than the Western medicine group and general massage group (P<0.05); scores of diarrhea, loose stool, lassitude, lusterless complexion, and appetite reduction decreased compared with before treatment (P<0.05); after treatment, all integrals of treatment group were lower than Western medicine group and general massage group (P<0.05). The total effective rates of the treatment group, Western medicine group and general massage group were 92.50％ (37/40), 72.50％ (29/40), and 75％ (30/40) respectively. There was statistical significance among the three groups (P<0.05). Conclusion Mingmen eight array massage combined with compound lysine hydrochloride and zinc gluconic granules can effectively alleviate the clinical symptoms of patients with infantile diarrhea, and improve the level of laboratory parameters.

A new strain of Streptomyces regensis was isolated from soil to produce a novel antibiotic AGPM possessing a strong antitumor activity In order to study on the metabolic path of the novel antibiotic AGPM, the protein patterns from the strain of Streptomyces regensis at different culture period were analyzed by using two eimensional polyacrylamide gel electrophoresis Comparing with sample from growth phase, seventeen new protein spots were found in that from antibiotic production phase The results demonstrated that the special proteins might be related with the antibiotic AGPM biosynthesis from Streptomyces regensis

Objective To evaluate the potential use of a probe melting analysis (PMA) assay in detecting the embB mutations which confer resistance against ethambutol in Mycobacterium tuberculosis. Methods The analysis sensitivity and specificity of PMA were investigated by detecting a serially diluted H37 Rv DNA and a reference panel from National Institute for the Control of Pharmaceutical and Biological Product. Six hundred and thirteen sputum samples were collected from the Xiamen Center for Disease Control and Prevention, Xiamen First Hospital and Center for Zhangzhou Disease Control and Prevention from September 2009 to April 2010. The PMA assay was then evaluated by detecting 613 clinical isolates and the results were compared with the sequencing results. Results The PMA assay could specifically detect Mycobacterium tuberculosis and had a limit of detection of 3 copies per reaction. The assay results with 613 clinical isolates showed that PMA gave a 100% concordance with sequencing in the 583 qualified samples, among which 34 were mutations at embB 306,23 at embB 378-380, 3 at embB 406 and 3 at embB 497. Conclusions PMA assay is a sensitive and specific method enabling efficient detection of common embB mutations causing ethambutol-resistance. The rapidness of this method together with its reliability would facilitate its use in routine testing.

Objective To explore the risk factors for clinical anastomotic leakage after resection of rectal cancer in China. Methods By meta-analysis we made a comprehensive analysis of the risk factors for clinical anastomotic leakage after resection of rectal cancer based on 19 articles published in China between January 1999 and January 2009. Results The anastomotic leakage rate was higher in the patients aged 60 years old and above than in those younger, with the combined odds ratio (OR) value being 0.50 (95% CI: 0.33-0.76) (P<0.01). The incidence rate was higher in the male patients than in the female ones, with the combined OR value being 2.17 (95% CI: 1.38-3.42) (P<0.01). The incidence rate in the patients with the distance of tumor from the lower margin to anal verge being 7cm and shorter was higher than that with longer distance, with the combined OR value being 1.79 (95% CI: 1.37-2.35) (P<0.01). The incidence rate in the patients who had received radiotherapy preoperatively was higher than that in those who had not, with the combined OR value of 3.66 (95% CI: 2.19-6.09) (P<0.01). The incidence rate in the patients who had received stapler anastomosis was higher than that in the patients who had received manual anastomosis, with the combined OR value being 0.70 (95% CI: 0.47-1.05), but there was no significant difference between them (P>0.05). The incidence rate was higher in the patients with diabetes mellitus than in the healthy ones, with the combined OR value being 3.16 (95% CI: 2.27-4.39) (P<0.01). The incidence rate was lower in the patients with Dukes A and B stages than in those with Dukes C and D stages, with the combined OR value being 0.61 (95% CI: 0.45-0.83) (P<0.01). The incidence rate in the patients with high malignance degree in clinicopathological types was higher than that with low malignance degree, with the combined OR value being 2.17 (95% CI: 1.38-3.42) (P<0.01). The incidence rate was lower in the patients who had received preventive colostomy than in those who had not, with the combined OR value being 0.39 (95% CI: 0.14-1.05), but there was no significant difference between them (P>0.05). The incidence rate was higher in the patients who had got selective operation than in those who had got emergency operation, with the combined OR value being 0.27 (95% CI: 0.13-0.56). Conclusion The risk factors of anastomotic leakage after resection of rectal cancer are as follows: 60 years old and above, male patients, diabetes mellitus, preoperative neo-adjuvant radiotherapy, the distance of tumor from the lower margin to the anal verge being shorter than 7cm, Dukes C and D stages, high malignance degree in clinicopathological types, and emergency operation.

Objectives To investigate the correlation between the level of peripheral blood inflammatory cytokines and kainic acid-induced seizure severity in rats.Methods 140 rats were divided into control and model group randomly,70 rats in each group.Model group rats were injected with kainic acid (10 mg/kg) by intraperitoneal,and the control rats were injected with sodium chloride.The change of their behaviors was observed and the concentrations of TNF-α,IL-1β,IL-4 and IL-10 were determined by ELISA in each group at different times.Results The rats showed epilepsy grand mal in 3 h-9 h after KA injection.The concentrations of TNF-α and IL-1β in model groups were significantly higher than those in control group in 6 h-12 h (6 h:(21.5±3.2) pg/ml vs (12.3±3.1)pg/ml;12 h:(20.6±4.2)pg/ml vs (11.5±3.8)pg/ml)(P＜0.05) and IL-4 in model group was higher at only 12 h ((53.55±3.08) pg/ml vs (33.26±4.16)pg/ml) (P＜0.05).The level of IL-10 in model groups was not statistically significant compared with control group (P＞0.05).Conclusion The proinflammatory cytokines (TNF-α and IL-l β) participate in the seizure procedure,meanwhile their levels and seizure severity have eminent correlations,but antiinflammatory cytokines (IL-4 and IL-10) had not.

Objective To discuss the clinical effects of hemiarthroplasty for the treatment of intertrochanteric comminuted fracture in aged patients.Methods From June 2006 to June 2010,91 cases with intertrochanteric comminuted fractures were treated by hemiarthroplasty using Zweymtüller long-stem cementless implant.There were 35 males and 56 females,with an average age of 81.2 years (range,75-94 years).There were fresh fractures in 75 cases and old fractures in 16 cases.According to classification of Evans-Jensen,there were 36 cases with type Ⅲ fracture and 55 with type Ⅳ.Osteoporosis was graded by Singh index,and there were 51 cases with level Ⅲ and 40 with level Ⅱ.The interval between injury and operation was 2-10 d (average,3.5 d).Results The surgery lasted 30-50 min (average,39 min).The blood loss was 320-610 ml (average,405 ml).The hospital stay was 14-21 d (average,16.5 d).Two cases who experienced with the infection of the respiratory system after operation,recovered through routine treatment.Thirty-seven cases were followed up for 15 to 38 months (average,24.5 months).According to the Harris score,the overall rate of excellent or good result was 83.8％ (31/37).Conclusion Hemiarthroplasty using Zweymüller long-stem cementless prosthesis is reliable option for intertrochanteric comminuted fracture in the aged patients.