Objective To investigate the effect on the differentiation of bone marrow mesenchymal stem cells (BMSC) with non-contact co-culture with mechanical stimulated ligament fibroblasts. Methods A cyclic 10% uniaxia strain at 1 Hz was applied on rat pelvic ligament fibroblasts, then were co-cultured with BMSC for 3, 6 and 12 days in non-contact condition. The protein expression of collagen Ⅰ ,Ⅲ in BMSC were detected by SP method and revealed by the mean gray value. The mRNA expressions of collagens type Ⅰ and type Ⅲ in the BMSCs were measured with real-time (RT)-PCR ,and the results were indicated by the ratio between the mRNA and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) . Results (1) Protein expression; after 3 days co-culture with pelvic ligament fibroblasts, expression of collagen Ⅰ and Ⅲ in BMSC are 82. 4 ± 3. 4 and 76. 8 ± 2. 5. When compared with 80. 2 ± 2. 6 and 74. 6 ± 1. 1 in BMSC without co-culture, there was no significant difference (P > 0. 05) . After 6 days co-culture with pelvic ligament fibroblasts, the expression of collagen Ⅰ and Ⅲ of 126. 6 ±2. 2 and 118. 6 ± 1. 4 in BMSC were significantly higher than 82. 7 ±3. 0 and 76. 2 ± 1. 3 in BMSC without co-culture (P < 0. 05). Similarly, after 12 days co-culture with pelvic ligament fibroblasts, the expression of collagen Ⅰ and Ⅲ of 135. 3 ±3. 4 and 128. 7 ± 2. 6 in BMSC were significantly higher than 86. 6 ± 1. 3 and 81. 8 ± 1.4 in BMSC without co-culture (P <0.05). (2)mRNA expression:after 3 days co-culture with pelvic ligament fibroblasts , the mRNA expression of type Ⅰ and type Ⅲ collagens in BMSC are 2. 10 ±0. 20 and 1. 20 ±0. 30. When compared with mRNA expression of 2. 01 ±0. 12 and 1. 13 ±0.21 in BMSC without co-culture, no significant difference were observed (P > 0. 05). After 6 days co-culture with pelvic ligament fibroblasts , the mRNA expressions of type Ⅰ and type Ⅲ collagens mRNA were 5. 60 ±0. 21 and 2. 61 ±0. 20, which were significantly higher than 3. 70 ±0. 33 and 1. 82 ± 0. 14 in BMSC without coculture (P < 0. 05). After 12 days co-culture with pelvic ligament fibroblasts, the mRNA expressions of type Ⅰ and type Ⅲ collagens of 5. 91 ±0.31 and 2. 92 ±0. 23 were significantly higher than 4. 04 ±0. 21 and 2. 04 ±0. 13 in BMSC without co-culture (P <0. 05). Conclusion Non-contact co-culture with mechanical stretch stimulated ligament fibroblasts, it might promote synthesis of types Ⅰ and Ⅲ collagen in rat BMSCs and induced BMSC differentiated into pelvic ligament fibroblasts.

Objective To understand the current situation and requirements of laboratory quality control of blood diseases in Chongqing .Methods A self‐designed questionnaire was designed ,and the investigation was carried out in 60 hospitals of Chongqing district .The main contents of the investigation were related to the instruments of work environment ,the regulations ,the conditions of the personnel ,etc .Results A total of 60 questionnaires were distributed ,57 questionnaires were valid .In the study ,only 14 .04%laboratories were the blood disease specialist laboratory ,most hospitals could complete the basic inspection items .66 .67% hospitals had all the experimental procedures and SOP documents .Most of staffs were with intermediate professional titles and undergraduate education .A total of 35 .08% experimental equipments with the value of over 5 million RMB ,45 .61% equipments had a special in‐spection records .About 45 .61% test reports would refer to the clinical performance ,and had a strict signing guideline .Most of the research objects reflected the problems ,including the backward equipments and the lack of personnel ,and hoped to set up telemedi‐cine indemnity system and carry out further education .Conclusion The overall quality of the laboratory for blood diseases in Chongqing is better ,it is recommended to strengthen the full quality control ,to establish emergency system ,and to improve the quality and level of laboratory .

Produced by bone cells and stored in the matrix.bone ceil growth factor can contribute to the regulation of bone growth.Gene regulation has its role in bone development.Many factors of this kind have been founded recently.They play their role during the bone formation and absorption processes in the way of autocrine and paracrine.The genes of core binding factor α_1(Cbfα_1)and estrogen receptor α(Erα)have been the recent years'hot factors that are related to the bone development,Gene polymorphism refers to variation on gene level which often occurs in the non-coding domain or the domain bearing no important regulatory function in gene order.This article reviewed the research status of the relation between Cbfα_1 and Erα gene polymorphism with bone development.

Adenocarcinoma, Follicular ; pathology ; surgery ; Aged ; Aged, 80 and over ; Carcinoma, Papillary ; pathology ; surgery ; Follow-Up Studies ; Humans ; Lymphatic Metastasis ; Middle Aged ; Neck Dissection ; Survival Analysis ; Thyroid Neoplasms ; pathology ; surgery ; Thyroidectomy ; methods

Object To develop an improved RP-HPLC method for determining ferulic acid in the human serum. Methods The determination was carried on RP-HPLC, using Kromasil-C 18column, methanol-water-acetic acid (36.4∶63∶0.6) as mobile phase with a flow rate of 1.0 mL/min and at the detection wavelength of 322 nm, and p-hydroxybenzaldehyde was used as internal standard. The serum sample was deproteinized with acetonitrile to extract ferulic acid. Results The calibration curve showed good linearity over the range of 9.94-159.04 ng/mL (r=0.992 5). RSD was less than 10% within day and day-to-day, the average recovery was 99.77% and the minimal concentration in serum was 5 ng/mL. Conclusion This method, which is simple, rapid, sensitive, reproducible and low toxic, is appropriate for the quantitative determination of ferulic acid in the human serum.

Objective To investigate the effects of lipopolysaccharide (LPS) on the expressions of sodium taurocholate co-transporting polypeptide (Ntcp) and bile salt export pump (Bsep),as well as the liver function markers in the serum including total bilirubin (TBIL),total bile acids (TBA),alanine aminotransferase (ALT),aspartate aminotransferase (AST) in mice.Methods One hundred and twenty-eight C57BL/6 mice were intra-peritoneally injected with different doses of 5,10,20 or 40 mg/kg LPS (n =24),respectively.No treatment or treated with 0.9％ NaC1 in mice as controls.Serum TBIL,TBA,ALT and AST levels were measured at 24 h,48 h and 72 h after LPS injection in each group.The mRNA expressions of Ntcp and Bsep were detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR).The liver histological sections were stained with haematoxylin and eosin (H&E).Results The Ntcp and Bsep mRNA expressions in mice liver were significantly lower in livers of LPS-treated mice within 24-72 h compared with control group,and the lowest level was reached at 24 h in a dose-dependent manner.And the relative expressions of Ntcp mRNA and Bsep mRNA were (0.64 ± 0.02),(0.53 ± 0.14),(0.25±0.09),(0.15±0.07)and (0.74±0.12),(0.58±0.11),(0.41±0.09),(0.27 ± ± 0.11) in livers of mice injected with LPS in the different doses of 5,10,20,40 mg/kg,respectively.In addition,serum levels of TBIL,TBA,ALT,and AST were significantly increased in mice of LPS-treated group compared with control group,particularly within 24 h after LPS treatment.Serum levels of TBIL,TBA,ALT,and AST were significantly decreased in mice of 40 mg/kg LPS-treated 72 h group compared with 24 h group presenting them with (1.29 ± 0.25) μ mol/L vs.(1.71 ± 0.22) μ moL/L,(6.97 ± 0.98) μmol/Lvs.(8.96±1.01) μmol/L,(120.17±21.08) U/L vs.(179.22±16.57) U/L,(360.34 ±35.31) U/L vs.(510.97 ± 34.70) U/L,respectively.Furthermore,histological changes in liver depend on dose and the course of LPS treatment.Cytoplasm rarefaction and inflammatory cells infiltration were detected at 24 h after treatment with 5 or 10 mg/kg LPS.Acidophilic and vacuolar degeneration,neutrophils infiltration in the hepatic sinusoid and portal area,the proliferation of bile ductulus were observed at 48 h,72 h after treatment with 5 or 10 mg/kg LPS.In the 20 or 40 mg/kg LPS treatment groups,focal necrosis,infiltration with inflammatory cells,proliferation of bile ductulus and expansion of duct were observed at 24 h,48 h and 72 h after LPS treatment.Conclusions LPS decreases the mRNA expressions of Ntcp and Bsep in a dose dependent manner in mice,contributing to mechanism of liver injury induced by endotoxin.

Objective To evaluate the value of two-dimension and color Doppler flow imaging(CDFI) in diagnosis of deep venous thrombosis(DVT) for different segments of lower extremity compared with digital subtract angiography(DSA). Methods Fifty-eight patients suspected of DVT enrolled in the study. Every segment venous of affected extremities was observed by CDFI. All patients underwent DSA examination. CDFI findings were compared with the results of DSA. Statistical analysis was according to quadruple tabular form. The indexes include sensitivity, specificity, accuracy, false positive rate and so on. Every segment of lower extremity deep venous was evaluated and analysis. Results Total of 150 vessels of fifty-eight patients were detected with CDFI and DSA. Evaluation findings of statistical analysis with quadruple tabular form as follows= sensitivity of common lilac vein was 82.6%, specificity was 66.7%; sensitivity of external lilac vein was 83.3%, specificity was 80.0%; sensitivity of hypogastria vein was 80.0 %, specificity was 66.7%;sensitivity of popliteal vein was 92.6 %, specificity was 92.2 % ;sensitivity of femoral vein was 96.2%, specificity was 93.8% ; sensitivity of anterior tibial vein was 71.4%, specificity was 75.0%; sensitivity of posterior tibial veins was 72.7%, specificity was 75.0%. Conclusions As the first-choice examination for DVT, ultrasonography has different diagnostic value for different segment of venous. It is more sensitive and accurate for femoral, popliteal and external iliac veins thrombus, but less sensitive for anterior,posterior tibial veins and bypass circuit vessels.

The antitumor effect and the mechanism of action of ursolic acid in A549 cells(human non-small-cell lung cancer cells) was investigated in this paper. Firstly, MTT assay was performed to test whether ursolic acid could inhibit the growth of A549 cells. Secondly, Western blot was utilized to measure the expression level of COX-2 and the activation of MAPKs. The MTI assay revealed that ursolic acid inhibited the growth of A549 cells. The result of Western blot suggested that ursolic acid inhibited the expression of COX-2 and activation of ERK and ERK specific inhibitor PD98059 suppressed the expression of COX-2 synergistically with ursolic acid. Taken together, our data suggest that ursolic acid can suppress LPS-induced COX-2 expression in A549 cells, which could be due to the inhibition of the activation of ERK.

Objective To detect the mutations of the marOR gene and study the relations with the expressing level of the acrAB-tolC efflux pump in Shigella. Methods marOR genes were amplified by PCR for 100 clinical isolates and 5 reference strains of Shigella. The PCR products were digested by restriction endonuclease Taq Ⅰ , then analyzed by single strand conformation polymorphism(SSCP). The marOR genes of the mutated strains and sensitive strain were sequenced and the expressing leveLs of acrA, acrB and talC were determined by RT-PCR. Susceptibility tests of tetracycline (TE), chloramphenicol (C), ampicillin (Am) , gentamycin (GM), norfloxacin (NOR) and selectrin (SMZ-TMP) were performed in sequenced strains. Results marOR genes were found in all strains detected. SSCP analysis found the rate of mutations in marOR genes was 23%. Among 11 marOR gene-mutated strains which were sequenced, there were 9 strains having a four-base absence and three single-base mutations in different loci. The expressing levels of the acrAB-tolC efflux pump in the 11 strains were higher than those in sensitive strains and reference strain. Furthermore the 11 strains were multi-drug resistance. Conclusion The mutation rate of marOR gene in Shigella was high and the acrAB-tolC efflux pump genes were over-expressive in marOR gene-mutated strains which were multi-antibiotic resistance in the study.

Objective To detect the influence of marOR mutations on antibiotic resistance in Shigella spp. Methods The marOR gene with four-base deletion was amplified by overlap PCR, then inserted in a T-vector and transformed into DH5α. The clone of marOR gene with four-base deletion and three point mutations was prepared from the strain having these mutations. Electrophoresis and sequencing were preformed to certify the correction of the cloned genes. Drug susceptibility tests were preformed for the strains harbouring the different clones [DH5α, DH5α (T), DH5α (marOR), DH5α (marOR-CATT), DH5α(marOR-CATT + 3m)]. Results Compared with the control strain (DH5α-T), the antibiotic resistances of marOR with four-base deletion [DH5α (marOR-CATT)] were higher to streptomycin, tobramycin, cefazolin and cefalexin, and the antibiotic resistances of marOR with four-base deletion and three point mutations [DH5α (marOR-CATT + 3m)] were higher to streptomycin and to tetracycline. The antibiotic resistances of DH5α (marOR-CATT) and DH5α (marOR-CATT +3m) to streptomycin, tetracycline, chloramphenicol, cefazolin, levofloxacin, ciprofloxacin and norfloxacin were higher than DH5α (marOR). The diameters of the antibiotics except the trimethoprim between DH5α (marOR-CATT) and DH5α (marORCATT +3m) had not significant disparity. Conclusion The four-base deletion in 1376-1379 sites of the marOR gene increased the resistance of Shigella spp to some antibiotics. The point mutations in 1411, 1417,1435 sites of the marOR gene have little influence on the antibiotic resistance of Shigella spp.