Objective To investigate the effects of pentoxifylline on the cell proliferation of, collagen synthesis and TGF-β1 expression by human fibroblasts derived from keloid. Methods Skin samples were obtained from the lesions of 3 patients with keloid and normal skin of 3 human controls followed by primary culture of fibroblasts. Fibroblasts of 5th to 8th generation were cultured with pentoxifylline of 0.1 to 3 g/L for various durations. Then, MTT assay was performed to detect the cell proliferation of fibroblasts, double antibody sandwich-enzyme linked immunosorbent assay (ELISA) to measure the expression of TGF-β1, and reversetranscription PCR to examine the mRNA expressions of procollagen Ⅰ and Ⅲ in these fibroblasts. Results The pentoxifylline of 0.1 to 2 g/L markedly inhibited the proliferation of fibroblasts derived from keloid lesions and normal skin, in a dose- and time-dependent manner, with the strongest effect observed in fibroblasts treated with pentoxifylline of 2 g/L. A significant reduction was induced in the TGF-β1 mRNA expression in keloidand normal skin-derived fibroblasts by pentoxifylline of 0.5 to 2 g/L (all P < 0.01), and in the mRNA expression of procollagen Ⅰ and Ⅲ by pentoxifylline of 1 and 2 g/L (P < 0.05 or 0.01). Concretely, the relative mRNA expression level of procollagen Ⅰ and Ⅲwas 0.873 ± 0.077, 0.571 ± 0.050 respectively in keloid fibroblasts respectively, and 0.473 ± 0.035, 0.370 ± 0.045 in the control fibroblasts, after treated with pentoxifylline of 1 g/L, 0.750 ± 0.036 and 0.433 ± 0.045 respectively in keloid-derived fibroblasts, 0.390 ± 0.030 and 0.250 ±0.123 respectively in the control fibroblasts, after treated with pentoxifylline of 2 g/L, significantly lower than that in the keloid-derived (1.216 ± 0.061 and 0.953 ± 0.060) and control (0.836 ± 0.080 and 0.776 ± 0.041) fibroblasts without treatment. Conclusion Pentoxifylline shows an evident suppressive effect on the cell proliferation of, as well as the expression of TGF-β1 and procollagen Ⅰ and Ⅲ in fibroblasts derived from keloid lesions and normal skin.

AIM:To determine the contents of sinomenine and chelidonine in Tong’an Injection(Caulis sinomenii, Chelidonium majus Linn, etc). METHODS: HPLC was used. The conditions included the gradient elution with methanol-0.1% triethylamine. The detection wavelength was set at 280 nm. RESULTS: The calibration curve was linear in the range of 162-1 620 ?g for the sinomenine and the range of 35-350 ?g for chelidonine, respectively. The average recovery for sinomenine was 99.56% and the relative standard deviation was 0.41%(n=5). The average recovery for chelidonine was 99.46% and the relative standard deviation was 0.62% (n=5). CONCLUSION: The method is simple, rapid and specialty. It can be used for the determination of sinomenine and chelidonine in Tong’an Injection.

Objective To explore the application range and preponderance of different therapeutic methods of traditional medicines in overseas countries with retrospective studies on bibliographies of overseas traditional medicines. Methods All bibliographies were articles written in August 1964 to September 2009 and came from MEDLINE data base through searching 17 inferior words of"replacement therapy" in MeSH subject headings. Results We got altogether 10103 articles in 1633 kinds of periodicals concerned with replacement therapy, and summed up to 9841 effective articles in the end. Conclusion More and more attentions have been paid to traditional medicine in overseas countries; Such kinds of articles were mostly published in American and British medical periodicals; articles with advanced technologies and high reliability have more chances to be adopted by medical periodicals; traditional Chinese medicine plays a leading role in the fields of traditional medicines in the whole world; replacement therapy were increasingly adopted to deal with disease protection and health care.

This study aimed to investigate whether β-elemene could improve the dysfunction of vascular endothelial cells induced by low shear force (LSS), and the proliferation and migration of vascular smooth muscle cells induced by oxidized low-density lipoprotein (ox-LDL). Parallel plate flow chambers and ox-LDL were used to establish vascular endothelial cells (ECs) dysfunction model and vascular smooth muscle cell (VSMCs) proliferation and migration model, respectively, and the effects of β-elemene on ECs dysfunction and VSMCs proliferation and migration were examined. The activity of ROS in ECs was measured by DHE and the activity of NO in ECs was tested by DAF-FM DA. The protein phosphorylation of Akt and ERK in ECs were detected by Western blot. The proliferation of VSMCs was measured by MTT. The migration of VSMCs was examined by cell scratch test and Transwell assay. The gene expression of MMP-2 and MMP-9 in VSMCs was measured by RT-qPCR. In ECs, β-elemene could significantly reduce the LSS-induced increase in ROS, significantly increase the LSS-induced decrease in NO, decrease the phosphorylation of ERK, and increase the phosphorylation of Akt. In VSMCs, β-elemene could significantly reduce the proliferation and migration of VSMCs induced by ox-LDL, and reduce the gene expression of MMP-2 and MMP-9. To conclude, β-elemene can improve the LSS-induced ECs dysfunction and ox-LDL-induced VSMCs proliferation and migration.

For decades,there have been no systematic studies on the reference intervals of Chinese populations.With the support of the Ministry of Health,the Chinese Society of Laboratory Medicine launches a research project investigating the Chinese reference intervals for common clinical laboratory tests.Up to now,the first batch of reference intervals for common liver function,electrolytes and blood cell analysis have been published in forms of healthy professional standards.Medical institutions are suggested to use these new reference intervals in clinical practice.

Objective To investigate the expression and significance of γH2AX in cervical squamous carcinoma.Methods Firstly,DNA were extracted from 74 cervical squamous carcinoma samples and PCR were tested for HPV infection.Secondly,formalin-fixed paraffin-embedded tissue sections (4 μm)were stained with H&E method to detect cervical lesions grading.Thirdly,HPV16 DNA were examined by in situ hybridization(ISH) and γH2AX,p16 were examined by immunohistochemical (IHC) staining.Then,30 cases typical tissue sections in which including the normal cervical tissue,cervical intraepithelial neoplasia and cervical carcinoma in situ were selected for comparing the HPV DNA loading,and the γH2AX and,pl6 expression.Finally,the feasibility of γH2AX serving as a biomarks in HPV infection-related cervical carcinogenesis were analyzed.Results In this study,HPV infection ratio is 98.65％,and HPV16 is the most common type with 74.32％ infection.In situ hybridization showed no HPV16 DNA exist in normal cervical tissues and CINI.In CIN Ⅱ HPV DNA exist mainly as episomal DNA.With the increasing of cervical lesions grade,HPV DNA was integrated into chromosome steadily.The expression of γH2AX and pl6 were positively associated with grading of cervical lesions.HPV DNA and γH2AX protein co-exist primarily in the prickle cell layer and the granular cell layer.The HPV DNA and p16 protein exist in different cell layer.Conclusion γH2AX may be employed as a biomarker for HPV positive cervical carcinogenesis.

Objective To verify and monitor the performance of accuracy, precision and comparability of 26 clinical biochemical analytes (29 methods) in the six centers involved in multi-centers reference intervals research, and to ensure the reliability of theirmeasurement results.Methods During the period of the systems evaluating, two levels of commercial quality control materials and fresh frozen human serum reference materials were applied to verify the performance of inter-laboratory precision and accuracy of analysis systems. During the period of samples testing, the commercial quality control materials were measured whenever samples were analysed, the fresh frozen serum reference materials were measured once a month.The coefficient of variations (CVs), bias and total errors were calculated to assess the precision, accuracy and comparability.Results Verification of precision and accuracy: ( 1 ) the ranges of CVs of 29 methods in the six laboratory laboratories were 0.4%-6.0%, the CVs of all 29 methods met the criterion . (2) The overall average bias of the analysis systems of 21 analytes (24 methods) ranged from -5.15%( ALT) to 4.46% ( Ur ) .Among 24 methods the overall average bias of TP, Glu-GOD, Ur, Cl, Ca exceeded the acceptable range.The quality assessment during the period of samples testing:(1) The overall average bias ranged from -1.95%(Ca) to 2.92%(Ur), median 1.26%, they all met the requirements of relevant standards.( 2 ) When commercial control materials were tested, the requirements of CVs were fulfilled for most methods in the six laboratories,and the CVs of TP, Alb, Cl, Ca exceeded the acceptable range.The overall average TE of all methods met the quality specification for the C-N controls material.For the C-P control material, only the overall average TE of TP (5.05%) exceeded thearceptable range while the other methods met the requirement in criterion.Conclusions The performance of precision and accuracy of the analysis systems used in the six laboratories passed the verification.During the period of sample testing, the performance of precision and accuracy of the most methods in the 6 laboratories met the requirements of quality specifications, and the overall performance was good.Because of the limitation of current technology the performance of some methods didn't fulfill the requirement of specifications, and need to be improved.

In recent years, artificial intelligence (AI) and oncology are rapidly integrated and developed, gradually moving towards clinical application. In China, the incidence and mortality of gastric cancer rank first among various tumors. The development of AI technologies such as convolutional neural network, narrow-band imaging magnifying endoscope, full-field digital section and survival circulation network has the potential to be applied to early screening, auxiliary diagnosis, pathological diagnosis, prognosis analysis, as well as individualized diagnosis and treatment process of gastric cancer. However, AI medical treatment still has many problems such as insufficient data sample size and difficulty in integration. Reasonable design and application of AI technology is expected to provide more scientific and accurate diagnosis and treatment for patients with gastric cancer, improve the overall survival rate and quality of life of patients with gastric cancer, and achieve more breakthroughs in the medical field.

This study was aimed to identify pET21b-HPV16E2/BL21(DE3) strain and to optimize the expression of human papillomavirus type 16 (HPV16) E2 protein by orthogonal analysis. Four influence factors on two levels were selected to increase the target protein quantity. The four factors were induction time, induction temperature, inductor concentration and cell density. The quantity of HPV16 E2 protein was used as the evaluation parameter. Induced by IPTG, HPV16 E2 protein was analyzed by SDS-PAGE and Western Blot. Target protein was analyzed by GIS imaging system to quantify the protein level. SPSS13. 0 software was applied to analyze the result. Data showed that the expression strain pET211rHPV16 E2/BL21(DE3) was identified correctly. HPV16 E2 protein expressed mainly at insoluble form. The 42KD protein band was identified by SDS-PAGE and Western blot. Orthogonal test was applied on influence factor analysis and expression optimization successfully. Main influence factors were inductor concentration and induction temperature. The optimimum condition of maximum expression quantity was 37 degrees C, 7h, 1.0 mmol/L IPTG and OD600 1.0. In this experiment, orthogonal test could not only be used to analyze the influential factors and promote the target protein expression, but also be used to provide a better experiment method for molecular biological study.
DNA-Binding Proteins ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; Human papillomavirus 16 ; metabolism ; Humans ; Oncogene Proteins, Viral ; biosynthesis ; genetics ; Papillomavirus Infections ; virology ; Recombinant Proteins ; biosynthesis ; genetics

Objective To investigate the expressions of transforming growth factor β1 (TGF-β1) and brain natrium peptide (BNP) in patients with diastolic heart failure (DHF),and to explore the correlation between plasma levels of TGF-β1,BNP and TGF-β1/BNP with parameter of diastolic function,diastolic dysfunction and New York Heart Association (NYHA) classification of cardiac function.Methods Hospitalized patients with DHF from October 2016 to November 2017 in Beijing Chaoyang Hospital were selected as subjects.At the same time,the age-and gender-matched non-heart failure hospitalized patients were selected as the control.The diastolic function index (E/e') was measured using cardiac ultrasound spectral Doppler and tissue Doppler methods.The diastolic dysfunction classification was evaluated according to the American Society of Echocardiography guidelines.Cardiac function was evaluated with NYHA classification.The levels of plasma TGF-β1 and BNP were measured with method of enzyme linked immunosorbent assay (ELISA).The correlation between the indicators was analyzed and the receiver operating characteristic (ROC) curve was drawn.Results A total of 186 patients were enrolled,including 114 patients as DHF group [54 males and 60 females,mean age (70.75 ± 11.45) years old] and 72 cases as control group [41 males and 31 females,mean age (68.74 ± 10.86) years old].The levels of TGF-β1 [(77.68 ± 42.31) ng/L] and BNP [(1 153.84 ± 564.96) ng/L] in patients with DHF were significantly higher than those of the control group [(18.76 ± 13.70),(264.07 ± 179.43) ng/L,t =15.62,13.77,P ＜ 0.01].Pearson correlation analysis showed that level of plasma TGF-β1 had a significant liner correlation with index E/e' (r =0.582,P ＜ 0.01),level of plasma BNP had a low-degree liner correlation with index E/e' (r =0.261,P ＜ 0.01),and TGF-β1/BNP had no correlation with index E/e' (r =0.081,P ＞ 0.05).Spearman correlation analysis showed that the levels of TGF-β1 and BNP were significantly correlated with diastolic dysfunction grading (r =0.473,0.417,P ＜ 0.01),while TGF-β1/BNP had no correlation with diastolic dysfunction grading (r =0.062,P ＞ 0.05).Plasma TGF-β1 and BNP had low-degree correlation with NYHA classification of heart failure (r =0.309,0.326,P ＜ 0.01),TGF-β1/BNP had no correlation with NYHA classification of heart failure (r =0.011,P ＞ 0.05).Logistic analysis showed that both plasma TGF-β1 and BNP were independent predictors of DHF (OR =1.264,2.283,P ＜ 0.05 or ＜ 0.01).The area under ROC curve (AUC) of BNP for prediction of DHF was 0.937 ± 0.064,and TGF-β1 was 0.597 ± 0.042.AUC areas of BNP and TGF-β1 were significantly different (P ＜ 0.01).Conclusions The expressions of plasma TGF-β1 and BNP in patients with DHF are higher than those without DHF.The levels of plasma TGF-β1 and BNP are significantly correlated with index E/e',diastolic dysfunction grading and NYHA classification.Both elevated BNP and TGF-β1 levels are independent predictors of DHF.Both plasma BNP and TGF-β1 have auxiliary diagnostic value on DHF and the diagnostic value of plasma BNP is greater than plasma TGF-β1.