The Role of MMP-9 in Regulating Degradation of the Splenic Microvascular Basement Membrane Induced by High Altitude Hypoxia / Rol de la MMP-9 en la Regulación de la Degradación de la Membrana Basal Microvascular Esplénica Inducida por Hipoxia de Altura

SUMMARY: To investigate changes of MMP-9 in the rat spleen and hypoxia-induced microvascular basement membrane under high altitude hypoxia. Thirty male specific pathogen-free Sprague Dawley rats were randomly divided into control and hypoxia groups, with 15 rats in each group. The rats in the control group were placed in Dingxi City, Gansu Province (2080 m above sea level) for 30 days. Rats in the hypoxia group were raised in a hypoxic environment in Maduo County, Qinghai Province (4300 m above sea level), for 30 days to establish a hypoxic rat model. Routine blood tests, MMP-9 mRNA, MMP-9 protein, and the spleen microvascular basement membrane were detected. (1) Compared with the control group, the red blood cell count, hemoglobin, and hematocrit levels of the rats in the hypoxia group were all increased; thus, a hypoxia model was successfully established. (2) Compared with the control group, the expression of MMP-9 mRNA and protein was significantly higher in the spleen of rats in the hypoxic group, and the difference was statistically significant (P <0.05). (3) Compared with the control group, the blood vessel basement membrane in the spleen of the hypoxia group was degraded. Under natural low air pressure and high altitude conditions, the expression of MMP-9 in rat spleen tissue increases and participates in the degradation of the microvascular basement membrane.

El objetivo de este trabajo fue investigar los cambios de la MMP-9 en el bazo de la rata y la membrana basal microvascular inducida bajo hipoxia a gran altura. Treinta ratas macho Sprague Dawley, libres de patógenos específicos, se dividieron aleatoriamente en dos grupos de 15 ratas cada uno, un grupo control y un grupo hipoxia. Durante 30 días las ratas del grupo control estuvieron en la ciudad de Dingxi, provincia de Gansu (2080 m sobre el nivel del mar). Las ratas del grupo de hipoxia se criaron en un entorno hipóxico en el condado de Maduo, provincia de Qinghai (4300 m sobre el nivel del mar), durante 30 días para establecer un modelo de rata hipóxica. Se realizaron análisis de sangre de rutina, ARNm de MMP-9, proteína MMP-9 y de la membrana basal microvascular del bazo. En comparación con el grupo control, el recuento de glóbulos rojos, la hemoglobina y los niveles de hematocrito de las ratas del grupo de hipoxia aumentaron; por lo tanto, se estableció con éxito un modelo de hipoxia. En comparación con el grupo control, la expresión de ARNm y proteína de MMP-9 fue significativamente mayor en el bazo de las ratas del grupo hipóxico, siendo la diferencia estadísticamente significativa (P <0,05). En comparación con el grupo control, la membrana basal de los vasos sanguíneos estaba degradada en el bazo del grupo hipoxia. En condiciones naturales de baja presión atmosférica y gran altitud, la expresión de MMP-9 en el tejido del bazo de la rata aumenta y participa en la degradación de la membrana basal microvascular.

Analysis of the clinical characteristics of 69 cases of occupational melanosis / 中国职业医学

Objective To summarize the clinical characteristics of patients with occupational melanosis. Methods Diagnostic data of 69 patients with occupational melanosis was analyzed using retrospective analysis. Results The main occupational hazards for the 69 patients with occupational melanosis were coal tar, petroleum and its fractionated products, pigments and dyes and their intermediates, rubber additives and rubber products. The median length of occupational exposure and disease latency were 8.0 and 6.0 years, respectively, with a highly positive correlation between them (Spearman correlation coefficients=0.962, P<0.01). Skin lesions were mainly found on exposed areas such as the face-to-neck and limbs, prevalence of 94.2% and 75.4% respectively. And 78.3% of patients had skin lesion on more than two sites. The lesions were mostly in the form of irregular flakes (59.4%), with a gray-black color (44.9%). About 43.5% of patients experienced skin itching. Complete blood count, liver function, and kidney function were all within normal ranges. Skin biopsy results showed that epidermal hyperkeratosis, thinning of the spinous layer, liquefaction degeneration of basal cells, increased superficial dermal melanocytes, and infiltration of lymphocytes, histiocytes, and melanocytes around the blood vessels. Reflectance confocal microscopy (RCM) detection showed focal liquefaction degeneration of basal cells in the lesions, with a significant infiltration of melanocytes and inflammatory cells in the dermal papillae and superficial layers. Conclusion The primary target organ of occupational melanocytes is the skin, and no damage to other organs was identified thus far. Results from skin biopsies and RCM examinations can be used for differential diagnosis.

Survey on the application of external cardiopulmonary resuscitation in Chinese children with sudden cardiac arrest / 中华儿科杂志

Objectives: To investigate the current application status and implementation difficulties of extracorporeal cardiopulmonary resuscitation (ECPR) in children with sudden cardiac arrest. Methods: This cross-sectional survey was conducted in 35 hospitals. A Children's ECPR Information Questionnaire on the implementation status of ECPR technology (abbreviated as the questionnaire) was designed, to collect the data of 385 children treated with ECPR in the 35 hospitals. The survey extracted the information about development of ECPR, the maintenance of extracorporeal membrane oxygenation (ECMO) machine, the indication of ECPR, and the difficulties of implementation in China. These ECPR patients were grouped based on their age, the hospital location and level, to compare the survival rates after weaning and discharge. The statistical analysis used Chi-square test and one-way analysis of variance for the comparison between the groups, LSD method for post hoc testing, and Bonferroni method for pairwise comparison. Results: Of the 385 ECPR cases, 224 were males and 161 females. There were 185 (48.1%) survival cases after weaning and 157 (40.8%) after discharge. There were 324 children (84.2%) receiving ECPR for cardiac disease and 27 children (7.0%) for respiratory failure. The primary cause of death in ECPR patients was circulatory failure (82 cases, 35.9%), followed by brain failure (80 cases, 35.0%). The most common place of ECPR was intensive care unit (ICU) (278 cases, 72.2%); ECPR catheters were mostly inserted through incision (327 cases, 84.9%). There were 32 hospitals (91.4%) had established ECMO emergency teams, holding 125 ECMO machines in total. ECMO machines mainly located in ICU (89 pieces, 71.2%), and the majority of hospitals (32 units, 91.4%) did not have pre-charged loops. There were no statistically significant differences in the post-withdrawal and post-discharge survival rates of ECPR patients among different age groups, regions, and hospitals (all P>0.05). The top 5 difficulties in implementing ECPR in non-ICU environments were lack of ECMO machines (16 times), difficulty in placing CPR pipes (15 times), long time intervals between CPR and ECMO transfer (13 times), lack of conventional backup ECMO loops (10 times), and inability of ECMO emergency teams to quickly arrive at the site (5 times). Conclusion: ECPR has been gradually developed in the field of pediatric critical care in China, and needs to be further standardized. ECPR in non-ICU environment remains a challenge.

Repurposed benzydamine targeting CDK2 suppresses the growth of esophageal squamous cell carcinoma / 医学前沿

Esophageal squamous cell carcinoma (ESCC) is one of the leading causes of cancer death worldwide. It is urgent to develop new drugs to improve the prognosis of ESCC patients. Here, we found benzydamine, a locally acting non-steroidal anti-inflammatory drug, had potent cytotoxic effect on ESCC cells. Benzydamine could suppress ESCC proliferation in vivo and in vitro. In terms of mechanism, CDK2 was identified as a target of benzydamine by molecular docking, pull-down assay and in vitro kinase assay. Specifically, benzydamine inhibited the growth of ESCC cells by inhibiting CDK2 activity and affecting downstream phosphorylation of MCM2, c-Myc and Rb, resulting in cell cycle arrest. Our study illustrates that benzydamine inhibits the growth of ESCC cells by downregulating the CDK2 pathway.

Effect of Recombinant Human Thrombopoietin (rhTPO) on Long-term Hematopoietic Recovery in Mice with Acute Radiation Sickness and Relative Mechanism / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect and relative mechanism of Recombinant Human Thrombopoietin (rhTPO) on long-term hematopoietic recovery in mice with acute radiation sickness.@*METHODS@#Mice were intramuscularly injected with rhTPO (100 μg/kg) 2 hours after total body irradiation with 60Co γ-rays (6.5 Gy). Moreover, six months after irradiation, peripheral blood, hematopoietic stem cells (HSC) ratio, competitive transplantation survival rate and chimerization rate, senescence rate of c-kit+ HSC, and p16 and p38 mRNA expression of c-kit+ HSC were detected.@*RESULTS@#Six months after 6.5 Gy γ-ray irradiation, there were no differences in peripheral blood white blood cells, red blood cells, platelets, neutrophils and bone marrow nucleated cells in normal group, irradiated group and rhTPO group (P>0.05). The proportion of hematopoietic stem cells and multipotent progenitor cells in mice of irradiated group was significantly decreased after irradiation (P<0.05), but there was no significant changes in rhTPO group (P>0.05). The counts of CFU-MK and BFU-E in irradiated group were significantly lower than that in normal group, and rhTPO group was higher than that of the irradiated group(P<0.05). The 70 day survival rate of recipient mice in normal group and rhTPO group was 100%, and all mice died in irradiation group. The senescence positive rates of c-kit+ HSC in normal group, irradiation group and rhTPO group were 6.11%, 9.54% and 6.01%, respectively (P<0.01). Compared with the normal group, the p16 and p38 mRNA expression of c-kit+ HSC in the irradiated mice were significantly increased (P<0.01), and it was markedly decreased after rhTPO administration (P<0.01).@*CONCLUSION@#The hematopoietic function of mice is still decreased 6 months after 6.5 Gy γ-ray irradiation, suggesting that there may be long-term damage. High-dose administration of rhTPO in the treatment of acute radiation sickness can reduce the senescence of HSC through p38-p16 pathway and improve the long-term damage of hematopoietic function in mice with acute radiation sickness.

Establishment of AML Mouse Model by Transplantation of Hematopoietic Cells from MLL-AF9 Transgenic Mice / 中国实验血液学杂志

OBJECTIVE@#To establish a leukemia mouse model induced by transplantation of hematopoietic cells from mixed lineage leukemia (MLL)-AF9 transgenic mice so as to provide the basis for the mechanism research and drug screening of acute myeloid leukemia (AML).@*METHODS@#MLL-AF9 knock-in mice were bred and identified. When the mice developed leukemia, white blood cell (WBC) count in peripheral blood, flow cytometry and morphology method were analyzed to identify the disease. When the WBC count in peripheral blood was more than 100×10@*RESULTS@#The natural onset times of leukemia on MLL-AF9 knock-in mice were 22-28 weeks. The spleens of the transgenic mice enlarged and the bone marrow showed the immature forms of myeloid leukemia cells. Both the bone marrow and spleen cells highly expressed myeloid markers, CD11b and Gr-1. At least 0.5×10@*CONCLUSION@#The leukemia model of hematopoietic cell transplantation based on MLL-AF9 transgenic mice is successfully established, which can be used for the study of the pathogenesis and evaluation of therapeutic effect of AML.

Characterization and Antioxidant Activities of Polysaccharide Isolated fromLinggui Zhugan Tang / 中国实验方剂学杂志

Objective:To characterize the structure of polysaccharide isolated from Linggui Zhugan Tang（LGZGT）,including monosaccharide composition and functional group detection, investigate the difference of the antioxidant activities of crude polysaccharide（CP） and pure polysaccharide（PP）, and provide the basis for the quality evaluation of LGZGT by in vitro bioassay. Method:The average molecular weight of CP was analyzed by high performance gel chromatography（HPGPC）. Gas chromatography-mass spectrometry（GC-MS） and fourier transform infrared（FT-IR） were employed to determine the structure of the polysaccharide. The antioxidant activities of CP and PP samples were evaluated on the basis of 1-diphenyl-2-picrylhydrazyl（DPPH） radical scavenging activity and OH radical scavenging activity. Result:The total polysaccharide was composed of single peaks, with a molecular weight of 3 689 Da. It was mainly composed of arabinose, mannose, glucose, galactose and fructose with a molar ratio of 6.85∶1.00∶109.21∶1.04∶21.82. Among them，glucose and fructose were the predominant components. In addition， IR study indicated the presence of pyranose and anomeric configurations in glycan structure, with two stereoisomers of glycosidic bond (α-glycosidic bond and β-glycosidic bond). It was found that the total polysaccharide had the ability of scavenging DPPH and hydroxyl radicals, and the activity of crude polysaccharide was better than that of refined polysaccharide. It was found in antioxidant research that the total polysaccharide had the ability of scavenging DPPH and hydroxyl radicals, and the activity of CP was better than that of PP. Furthermore, LC-Q-TOF-MS was used to qualitatively analyze the other components in CP, which indicated that it was related to the adsorption of pentacyclic triterpenoids in Glycyrrhiza uralensis. Conclusion:The polysaccharides and pentacyclic triterpenoids in LGZGT are the material basis for the antioxidative effect of LGZGT. The antioxidative activity determined by in vitro bioassay can be used as an evaluation index for the overall quality control of LGZGT.

An investigation into the mechanisms underlying the regulatory effect of the E2F6 transcription factor on proliferation and metastasis of malignant melanoma cells through β-catenin signaling pathway / 中华皮肤科杂志

Objective:To determine the expression of the E2F6 transcription factor in human malignant melanoma tissues and cell lines, and to evaluate the effect of E2F6 on proliferation, migration and invasion of a malignant melanoma cell line A375.Methods:Frozen tissues and paraffin-embedded tissue sections were collected from 50 cases of cutaneous malignant melanoma and 30 cases of pigmented nevus in Department of Dermatology, Chongqing Traditional Chinese Medicine Hospital from January 2012 to December 2017. Quantitative reverse transcription-PCR (qRT-PCR) was performed to determine the mRNA expression of E2F6 in the malignant melanoma and pigmented nevus tissues, as well as in 7 malignant melanoma cell lines (HM, A375, WM451, WM35, SK-MEL-1, Hs-695T and MDA-MB-435s) and pigmented nevus cells, and immunohistochemical study and Western blot analysis were conducted to determine the protein expression of E2F6 and β-catenin in the malignant melanoma tissues. An E2F6-inhibiting plasmid and a control plasmid were separately transfected into A375 cells by using a liposome-mediated transfection method, and the E2F6 gene-knockdown efficiency was verified by qRT-PCR and Western blot analysis. Cell counting kit-8 (CCK8) assay, soft-agar plate cloning assay, Transwell migration and invasion assays and 3D cell culture assay were conducted to evaluate the effect of E2F6 gene knockdown on the proliferation, migration and invasion of A375 cells, flow cytometry was performed to detect the cell cycle and apoptosis rate, and Western blot analysis was conducted to determine the protein expression of total β-catenin, activated β-catenin, c-Myc and cyclin D1. The comparison between two groups was carried out by t test, the comparison among several groups by one-way analysis of variance, and multiple comparisons by least significant difference t test; Pearson correlation coefficient was used to analyze the correlation between E2F6 and β-catenin expression in cutaneous malignant melanoma. Results:The E2F6 mRNA expression was significantly higher in the 7 malignant melanoma cell lines than in the pigmented nevus cells (all P < 0.001). qRT-PCR showed that the relative mRNA expression of E2F6 was significantly higher in the cutaneous malignant melanoma tissues (0.000 55 ± 0.000 17) than in the pigmented nevus tissues (0.000 18 ± 0.000 09, t = 3.22, P < 0.001). Both the immunohistochemical study and Western blot analysis showed significantly increased E2F6 protein expression, but decreased β-catenin protein expression in the cutaneous malignant melanoma tissues compared with the pigmented nevus tissues (all P < 0.001). Correlation analysis showed that E2F6 protein expression was negatively correlated with β-catenin expression in the malignant melanoma tissues (immunohistochemical study: r = -0.56, Western blot analysis: r = -0.63, both P < 0.01). After knockdown of the E2F6 gene in A375 cells, the mRNA and protein expression of E2F6 was significantly lower in the E2F6 inhibition group than in the control group ( t = 3.38, 2.76 respectively, both P < 0.001). CCK8 assay showed that the cellular proliferative ability was significantly lower in the E2F6 inhibition group than in the control group ( t = 4.58, P < 0.01) 48 hours after transfection; soft-agar plate cloning assay showed that the colony-formation ratio was significantly lower in the E2F6 inhibition group than in the control group ( t = 2.26, P < 0.001) ; Transwell migration and invasion assays showed that the number of cells crossing the chamber was significantly lower in the E2F6 inhibition group (165 ± 23, 96 ± 11 respectively) than in the control group (376 ± 22, 315 ± 31, t = 3.14, 2.12, respectively, both P < 0.01) ; 3D cell culture assay showed that the cell morphology markedly changed, and the invasive pseudopodia disappeared in the E2F6 inhibition group. Flow cytometry revealed that the proportion of cells at G0-G1 phase and apoptosis rate were significantly higher in the E2F6 inhibition group than in the control group (both P < 0.001). Western blot analysis showed significantly decreased protein expression of β-catenin, activated β-catenin and its downstream target proteins c-Myc and cyclin D1, but significantly increased protein expression of P21 in the E2F6 inhibition group compared with the control group (all P < 0.001) ; additionally, the E2F6 inhibition group showed significantly decreased protein expression of epithelial-mesenchymal transition-related molecules vimentin and N-cadherin, but significantly increased expression of E-cadherin compared with the control group (all P < 0.001) . Conclusions:The E2F6 transcription factor is highly expressed in malignant melanoma. Knockdown of the E2F6 gene in A375 cells can inhibit cell proliferation, migration and invasion by antagonizing the β-catenin signaling pathway.

Effect of different holmium laser fiber on regional temperature during lithotripsy with pediatric ureteroscope / 中华泌尿外科杂志

In order to assess the thermal effect of different holmium laser fiber during lithotripsy with WOLF F4.5/6.5 pediatric ureteroscope, we established an impacted ureter calculi model. Under 100mmHg irrigation pressure, regional temperature of different holmium laser fiber with varied working time and power were recorded. We found that the regional temperature was related with laser fiber diameters, power and working time settings. With 550 μm laser fiber, laser firing time longer than 3 s or 365 μm laser fiber firing more than 6 s, regional temperature exceeded 42℃, which would bring thermal injury towards ureter and subsequently cause ureter stricture.

Impact of gene dosage on recombinant transglutaminase production of Pichia pastoris / 生物工程学报

Based on the rDNA sequence of Pichia pastoris, a multi-copy gene expression vector of transglutaminase (pPICZα-rDNA-mtg) was constructed and transformed to the host strain (pGAP9-pro/GS115) expressing pro peptide, to obtain the co-expression strain pro/rDNA-mtg (GS115). Real-time fluorescence quantitative PCR (qPCR) was used to analyze transglutaminase gene copy number in the 4 positive strains. We further studied the effect of gene copy on the enzyme production of recombinant Pichia pastoris as well as high-density fermentation of higher expression strain in a 3-L fermenter. The mtg copy numbers of the 4 positive strains were 2.21, 3.36, 5.72 and 7.62 (mtg-2c, mtg-3c, mtg-6c and mtg-8c), respectively, and the enzyme production capacity and protein expression level were mtg-3c>mtg-2c>mtg-6c>mtg-8c. Mtg-3c and mtg-6c of high-density fermentation had the highest enzymatic activity and enzymatic activity per unit wet weight in the supernatant of 3.12 U/mL, 52.1 U/g (wet weight) and 2.07 U/mL and 36.5 U/g (wet weight), respectively. In terms of enzyme activity per unit wet weight, mtg-3c is 1.4 times higher than that of mtg-6c. The activity of purified enzyme (mtg-3c) was up to 7.21 U/mL and the protein concentration was 437.2 μg/mL. By analyzing the effect of mtg copy number on the enzyme production of recombinant strains, mtg-3c is suitable for the co-expression of two genes (pro and mtg) in pro/rDNA-mtg, and its enzyme activity is related to higher protein secretion of the strain.

Analysis of Volatile Components of Different Processed Products of Curcuma wenyujin Radix by HS-SPME-GC-MS / 中国实验方剂学杂志

Objective:To analyze and compare volatile components in raw and fried products of Curcuma wenyujin radix, and find out the material basis for the differences in efficacy between them,in order to provide powerful support and reference for the rational use of C. wenyujin radix of different processed products and establish a more perfect quality evaluation system. Method:HS-SPME combined with GC-MS was used to analyze and compare volatile components and relative percentage in different processed products of C. wenyujin radix. Result:Totally 42 peaks were detected from the raw product of C. wenyujin radix,and 23 components were identified. Totally 59 peaks were detected from vinegar-baked C. wenyujin radix,and 34 components were identified. Totally 64 peaks were detected from wine-baked C. wenyujin radix,and 40 components were identified. Compared with C. wenyujin radix,22 kinds of new ingredients were produced. Compared with C. wenyujin radix,29 kinds of new ingredients were produced in wine-baked C. wenyujin radix. The composition and content of volatile components in different processed products of C. wenyujin radix were changed. Conclusion:The headspace solid phase micro-extraction method is simple in operation and fast in extraction speed,with no use of organic solvent, no pollutant in the environment and a low operation cost. The method using headspace solid phase micro-extraction combined with gas chromatography-mass spectrometry has a high stability and reliability. It is suitable for rapid analysis of volatile components between C. wenyujin radix and its different processed products. It can provide a scientific basis for the quality evaluation of C. wenyujin radix and its different processed products,and a scientific reference for the rational use of C. wenyujin radix resources.

Mechanism underlying the inhibitory effect of overexpression of tumor suppressor gene DKK3 on migration and invasion of human cutaneous malignant melanoma cells / 中华皮肤科杂志

Objective To investigate the expression of DKK3 in human cutaneous malignant melanoma cells and tissues,and to evaluate the effect of transfection with DKK3 gene on migration and invasion of a malignant melanoma cell line A375.Methods Western blot analysis was performed to measure the relative expression of DKK3 in human cutaneous melanoma cell lines HM,A375,WM451,SK-MEL-1,Hs-695T,MDA-MB-435s and WM35,as well as pigmented nevus tissues.Real-time fluorescence-based quantitative PCR (RT-PCR) was conducted to determine the mRNA expression of DKK3 in 58 melanoma tissues (including primary melanoma and metastatic melanoma) and 30 pigmented nevus tissues from Chongqing Traditional Chinese Medicine Hospital between August 2014 and June 2017.The pcDNA3.1 (+)-Flag-Vector (control group) and pcDNA3.1 (+)-Flag-DKK3 (transfection group) were transfected into A375 melanoma cells separately.RT-PCR and Western blot analysis were performed to verify the overexpression of DKK3,and to evaluate the effect of DKK3 overexpression on the expression of molecules related to the migration and invasion of melanoma cells.Cell scratch assay,Transwell migration and invasion assay were conducted to assess the effect of DKK3 on the migration and invasion of A375 cells.Statistical analysis was done by a two-sample t-test for comparisons between two groups,one-way analysis of variance (ANOVA) for intergroup comparison,and least significant difference (LSD)-t test for multiple comparisons with the SPSS 13.0 software.Results DKK3 protein was absent or lowly expressed in the human melanoma cell lines,but highly expressed in the pigmented nevus tissues.There were significant differences in the mRNA expression of DKK3 among the primary melanoma tissues (2-ΔΔCt:[0.325 ± 0.150] × 10-3),metastatic melanoma tissues ([0.142 ± 0.210] × 103) and pigmented nevus tissues ([0.634 ±:0.120] × 10-3,F =46.57,P ＜ 0.05).In addition,the mRNA expression of DKK3 was significantly lower in the metastatic melanoma tissues than in the primary melanoma tissues and pigmented nevus tissues (LSD-t =2.48,3.12,both P ＜ 0.05).After transfection with DKK3,cell scratch assay showed that the migration rate was significantly lower in the transfection group (22.11％ ± 5.11％) than in the control group (54.36％ ± 23.22％,t =2.36,P ＜ 0.001).Transwell migration and invasion assay revealed that the number of A375 cells crossing the Transwell chamber was significantly lower in the transfection group (265 ± 33,76 ± 18 respectively) than in the control group (429 ± 41,135 ± 21 respectively;t =1.24,1.35 respectively,both P ＜ 0.001).After overexpression of DKK3 in the A375 cells in the transfection group,the mRNA and protein expression of E-cadherin were up-regulated,while the mRNA and protein expression of N-cadherin,vimentin,matrix metalloproteinase 2 (MMP2),MMP7 and MMP11 were down-regulated compared with the control group.Conclusions The expression of DKK3 is down-regulated in the melanoma cell lines and tissues,and the migration and invasion of A375 cells are markedly inhibited by overexpression of DKK3.DKK3 may be a target for inhibiting the metastasis of cutaneous malignant melanoma.

Roles of gut microbiota in the occurrence and development of polycystic ovary syndrome / 上海交通大学学报（医学版）

Polycystic ovary syndrome (PCOS) is a gynecologic endocrine disorder with complex etiology and pathogenesis. Many women of reproductive age are influenced by this disease due to infertility. The pathogenesis of PCOS remains unclear despite increasing studies in recent years. It is generally accepted that insulin resistance, hyperandrogenism and abnormal follicle development play a pivotal role in PCOS. Gut microbiota becomes a research hotspot in the aspect of infectious, immune and metabolic diseases recently. Previous studies have found that gut microbiota could modulate the synthesis and secretion of insulin, and affect metabolism of androgen and follicle development, providing us a new idea for unravelling the pathogenesis of PCOS. Based on these researches, fecal microbiota transplantation may be a promising treatment in rectifying intestinal microecology imbalance and improving metabolism. This paper reviewed recent research advances in the roles of gut microbiota in the occurrence and development of PCOS.

Risk factors for gestational diabetes mellitus among multiparae / 上海交通大学学报（医学版）

Objective • To investigate the risk factors for gestational diabetes mellitus (GDM) among multiparae. Methods • Women who had two consecutive pregnancies records in the International Peace Maternity and Child Health Hospital from January 2012 to January 2017 were included into this study. The case group (116 cases) and control group (464 cases) were matched at the ratio of 1:4 according to the pre-pregnancy age in index pregnancy. Clinical characteristics, biochemical parameters including oral glucose tolerance test (OGTT) and lipid profiles were took into consideration by virtue of their medical records. Multivariate Logistic regression analysis was used to compute the adjusted odds ratio (aOR) and 95%CI so as to identify the risk factors. Results • Compared with the control group, the case group was associated with greater body mass index (BMI) change between pregnancies (aOR=1.35, 95% CI=1.07-1.69), greater postprandial 1 h glucose load (aOR=1.99, 95% CI=1.55-2.55) and 2 h glucose load (aOR=2.02, 95% CI=1.51- 2.70) at OGTT in index pregnancy, and greater first-trimester fasting plasma glucose (aOR=1.96, 95% CI=1.16-3.32), total cholesterol (aOR=1.37, 95% CI=1.06-1.77) and triacylglycerol (aOR=1.53, 95% CI=1.10-2.14) in subsequent pregnancy. Conclusion • The elevated BMI change between pregnancies, the abnormal glucose and lipid profiles persisting from index to subsequent pregnancy lead to the occurrence of GDM.

Problems and countermeasures of physician multi-sited license based on the Grounded Theory:A perspective of the doctors / 中国卫生政策研究

Objective:Based on Grounded Theory,the objective of this study is to explore the problems of Phy-sician multi-sited license in Zunyi, Guizhou Province from the perspective of doctors and putting forward the corre-sponding countermeasures. Methods:From January to July,2017,based on purposive sampling and Grounded Theo-ry,43 doctors from nine medical institutions in Zunyi of Guizhou Province were interviewed using semi-structural in-terview syllabus. With the help of open coding, axial coding, selective coding, multi-sited licensed Physician were integrated to explore existing problems, and putting forward corresponding countermeasures. Results:After three-grade coding,105 concepts,17 categories,7 main categories and 5 core categories of multi-sited licensed Physicians were concluded. Then, 5 lines of Physician multi-sited license were formed, including development prospect and doctors aspiration, the factors that could stimulate development, development difficulties, relative supporting poli-cies,service needs and practice condition. Conclusion:The majority of doctors are willing to carry out multi-sited prac-tice,the relevant laws and regulations,the specific implementation details,salary distribution system and the improve-ment of treatment scheme of medical negligence,the support of medical institutions,the transformation of medical insti-tutions management,the necessary medical conditions that the second place of practice should provide,the distance of second place of practice which should be fully considered,the convenience of traffic conditions and corresponding team-work support are necessary requirements to be taken into consideration to carry out physician multi-sited license.

Risk factors for gestational diabetes mellitus among multiparae / 上海交通大学学报（医学版）

Objective·To investigate the risk factors for gestational diabetes mellitus (GDM) among multiparae. Methods?·?Women who had two consecutive pregnancies records in the International Peace Maternity and Child Health Hospital from January 2012 to January 2017 were included into this study. The case group (116 cases) and control group (464 cases) were matched at the ratio of 1:4 according to the pre-pregnancy age in index pregnancy. Clinical characteristics, biochemical parameters including oral glucose tolerance test (OGTT) and lipid profiles were took into consideration by virtue of their medical records. Multivariate Logistic regression analysis was used to compute the adjusted odds ratio (aOR) and 95%CI so as to identify the risk factors. Results?·?Compared with the control group, the case group was associated with greater body mass index (BMI) change between pregnancies (aOR=1.35, 95%?CI=1.07-1.69), greater postprandial 1 h glucose load (aOR=1.99, 95%?CI=1.55-2.55) and 2 h glucose load (aOR=2.02, 95%?CI=1.51-2.70) at OGTT in index pregnancy, and greater first-trimester fasting plasma glucose (aOR=1.96, 95%?CI=1.16-3.32), total cholesterol (aOR=1.37, 95%?CI=1.06-1.77) and triacylglycerol (aOR=1.53, 95%?CI=1.10-2.14) in subsequent pregnancy. Conclusion?·?The elevated BMI change between pregnancies, the abnormal glucose and lipid profiles persisting from index to subsequent pregnancy lead to the occurrence of GDM.

Roles of gut microbiota in the occurrence and development of polycystic ovary syndrome / 上海交通大学学报（医学版）

Polycystic ovary syndrome (PCOS) is a gynecologic endocrine disorder with complex etiology and pathogenesis.Many women of reproductive age are influenced by this disease due to infertility.The pathogenesis of PCOS remains unclear despite increasing studies in recent years.It is generally accepted that insulin resistance,hyperandrogenism and abnormal follicle development play a pivotal role in PCOS.Gut microbiota becomes a research hotspot in the aspect of infectious,immune and metabolic diseases recently.Previous studies have found that gut microbiota could modulate the synthesis and secretion of insulin,and affect metabolism of androgen and follicle development,providing us a new idea for unravelling the pathogenesis of PCOS.Based on these researches,fecal microbiota transplantation may be a promising treatment in rectifying intestinal microecology imbalance and improving metabolism.This paper reviewed recent research advances in the roles of gut microbiota in the occurrence and development of PCOS.

Monomorphic epitheliotropic intestinal T-cell lymphoma: A clinic-pathologic study of 3 cases and literature review / 解放军医学杂志

Objective To summarize the clinicopathological features of monomorphic epitheliotropic intestinal T cell lymphoma (MEITL),and provide the experiences and lessons for diagnosis and treatment.Methods The clinical manifestations,histopathological and immunohistochemical characteristics,diagnosis,treatment and prognosis of 3 cases of MEITL were analyzed retrospectively.Meanwhile,the other cases published in the literatures were reviewed and summarized.Results Two cases of middle aged and aged men and one young woman were reported in present paper.These lesions of all the 3 patients were at the small intestine,of which the female patients had skin and mammary gland involvement.The clinical manifestations were abdominal distention,mulligrubs and intestinal perforation,and the main pathological feature was the diffuse infiltration of the homogeneous tumor cells in the whole layer intestinal wall.Immunohistochemical examination showed that tumor cells expressed all T cell markers CD3,CD8,CD56,T cell cytoplasmic antigen (TIA-1),CD4 and CD5,and Ki-67 proliferation index was 50％-70％.As to the treatment,case 1 received CHOP and GDP regimens successively,and died 2 years after final diagnosis.Case 2 received CHOP regimen for 1 course of chemotherapy and died of giving up treatment.Case 3 gave up treatment,voluntary discharged and then lost.It is clear according to the previous literatures and our experiences that the MEITL possesses high invasiveness and low incidence,and makes the diagnosis difficult;the patient's response to the current treatment is poor.Conclusions MEITL is prone to be in the small intestine.The diagnosis of MEITL should be based on clinical manifestations,pathological features and immunohistochemical detection results.MEITL is rare,the course of disease progresses rapidly,and the patients have the tendency to have intestinal perforation and poor prognosis.Early diagnosis and positive chemotherapy may lead to a better prognosis.

Fluacrypyrim induced apoptosis in acute myeloid leukemia cell lines / 国际药学研究杂志

Objective To investigate the effect of fluacrypyrim on the induction of apoptosis in human acute myeloid leuke-mia(AML)cell lines,NB4,THP-1 and HL-60,and explore the related mechanisms.Methods Trypan blue dye exclusion assay was used to estimate the growth of NB4,THP-1,and HL-60 cells after treatment with various concentrations of fluacrypyrim(1.25, 2.5,5 and 7.5 μmol/L)for 72 h.Cell apoptosis was evaluated by AnnexinⅤ-FITC/PI double stainning for the NB4 and THP-1 cells treated with fluacrypyrim(1.25,2.5 and 5 μmol/L)for 48 h as well as the HL-60 cells treated with fluacrypyrim(2.5,5 and 7.5 μmol/L) for 72 h.Western blotting was used to examine the protein expression of apoptotic regulators Bax,Mcl-1 and Caspase 3 in the NB4 cells treated with fluacrypyrim(1.25,2.5 and 5 μmol/L)for 24 h.Then,NB4 Cells were pretreated with Caspases inhibitor benzyloxy-carbonyl-Val-Ala-Asp-fluoromethylketone(Z-VAD-FMK)and exposed to fluacrypyrim at 2.5 μmol/L for 24 h,which was then evaluat-ed for the apoptosis using AnnexinⅤ-FITC/PI double stainning.Western blotting was used to examine the expression of the phosphory-lated and total proteins of mitogen-activated protein kinase(MAPK)signaling molecules,ERK,JNK and P38,in the NB4 cells treat-ed with fluacrypyrim(1.25,2.5 and 5 μmol/L)for 24 h. NB4 Cells were pretreated with ERK inhibitor U0126,JNK inhibitor SP600125,or P38 inhibitor SB203580 for 1 h and then exposed to fluacrypyrim at 2.5 μmol/L for 24 h,which was then analyzed for the apoptosis by the AnnexinⅤ-FITC/PI double stainning.Results The proliferation of NB4,THP-1 and HL-60 cells was inhibited by the treatment with fluacrypyrim(2.5,5 and 7.5 μmol/L)for 72 h.The apoptosis induced in the NB4 and THP-1 cells by the fluacry-pyrim treatment at 5 μmol/L for 48 h and in the HL-60 cells by the fluacrypyrim treatment at 7.5 μmol/L for 72 h were significant as compared with the control group(P<0.01).Mechanically,fluacrypyrim at the concentrations of 2.5 and 5 μmol/L effectively up-regu-lated the expression of Bax(P<0.01 and P<0.05)for the 2.5 and 5 μmol/L,respectively,down-regulated the expression of Mcl-1 (P<0.01)and activated Caspase 3(P<0.01)in the NB4 cells when compared with the control group(P<0.01).The pretreatment of the NB4 cells with Z-VAD-FMK blocked the apoptosis induced by fluacrypyrim.Furthermore,the fluacrypyrim(2.5 and 5 μmol/L) treatment increased the ERK,JNK and P38 phosphorylation(P<0.01),while the pretreatment of the NB4 cells with U0126 signifi-cantly inhibited the the fluacrypyrim-induced apoptosis(P<0.01),as compared with the control group.Conclusion Fluacrypyrim effectively inhibits the cell proliferation and induces caspase-dependent cell apoptosis in AML cells.Activation of ERK/MAPK signal-ing pathway might play an important role in the action of fluacrypyrim.

Clinical effect of fire needle in treating vitiligo and its CLSM image characteristics / 重庆医学

Objective To evaluate the clinical effect of fire needle in treating vitiligo and the characteristics of vitiligo image by the confocal laser scanning microscope(CLSM).Methods The randomized self-controlled experiment design was adopted.Each patient selected two symmetric or adjacent white patches and randomly received the fire needle treatment or tacrolimus treatment. The duration of treatment was 3 months.The CLSM images of white patches were recorded before treatment and after 3,6 times of fire needle treatment.Results Among 41 cases of stable stage vitiligo,The effective rates of the fire needle group and tacrolimus group were 82.9% and 78.0% respectively,and the difference was not statistically significant(P>0.05).After fire needle treat-ment,dentritic melanin cells appeared,the pigment granules gradually appeared around the basal layer and corpora papillare,and formed the pigment ring.Conclusion Fire needle and tacrolimus have the similar effect in treating vitiligo,moreover CLSM can be used as the non-invasive,objective and reliable detection means of the recovery of vitiligo melanocyte.