OBJECTIVE To investigate the effect and mechanism of α7 nicotinic acetylcholine receptor （α7nAChR） agonists PNU-282987 on cognitive function in temporal lobe epilepsy （TLE） model rats. METHODS Sixty rats were randomly divided into control group， model group， PNU-282987 group （3 mg/kg） and methyllycaconitine （MLA）+PNU-282987 group （6 mg/kg MLA+3 mg/kg PNU-282987）， with 15 rats in each group. Except for control group， the TLE model was established in the other groups. After the model was successfully established， each group was given relevant medicine or normal saline intraperitoneally， once a day， for two consecutive weeks. The epilepsy attack of rats was observed and scored， and the duration of seizures was recorded； the cognitive function of rats was detected； pathological morphology of neurons in CA1 region was observed； the levels of tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6） and IL-1β in the hippocampus were detected； the positive expressions of ionized calcium-binding adapter molecule-1 （IBA-1）， α7nAChR， nuclear factor-κB （NF-κB） p65， p-NF-κB p65 in the hippocampus were detected. RESULTS Compared with model group， the score and duration of seizures， the number of IBA-1 positive cells， the levels of TNF- α， IL-6 and IL-1β， the expressions of NF- κB p65 and p-NF- κB p65 protein decreased significantly in the hippocampus （P＜0.05）； the escape latency time was shortened significantly （P＜0.05）， the time spent in the original platform quadrant and times of crossing the platform increased significantly （P＜0.05）； neuronal damage in the CA1 region of the hippocampus was significantly reduced； the expression of α7nAChR protein increased significantly in hippocampus （P＜0.05）. Compared with PNU-282987 group， the above indexes of rats in MLA+PNU-282987 group were reversed significantly （P＜0.05）. CONCLUSIONS PNU-282987 could improve cognitive dysfunction in TLE model rats， and its mechanism may be associated with inhibiting microglia-mediated inflammatory response through α7nAChR/NF- κB signaling pathway， thus reducing hippocampal neuronal damage.

Objective:To explore the differential expression profile of miRN in the development of diabetes nephropathy (DN), and further explore the mechanism of miR-126-5p involved in high glucose induced injury of renal tubular epithelial cells.Methods:Firstly, we downloaded existing chip data from the Gene Expression Integrated Database (GEO) and used GEO2R, miRanda, gene ontology (GO) analysis, and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis to mine differential miRNAs. Subsequently, a high glucose induced HK-2 cell injury model was used and divided into three groups: high glucose model group, si-HOTAIR group, and si HOTAIR+ miR-126-5p inhibitor group. The three groups of cells were sequentially transfected with siRNA-NC, siRNA-HOTAIR, and siRNA-HOTAIR+ miR-126-5p mimic, and cultured in a medium containing 60 mmol/L glucose. Flow cytometry was used to detect changes in apoptosis levels in each group, while cell counting kit-8 (CCK-8) was used to detect changes in cell proliferation.Results:Through data mining analysis using GEO, it was found that compared to ordinary mice, DN mice had 74 upregulated miRNAs and 80 downregulated miRNAs in their kidney tissue. Enrichment analysis results showed that miRNAs could target signaling pathways such as Wnt, PKG, MAPK, and Rap1, and miR-126-5p was significantly downregulated. In the high glucose induced HK-2 cell injury model, the experimental results showed that the inhibitory effect on cell proliferation activity was more significant at a high glucose concentration of 60 mmol/L ( P<0.05); High glucose stimulation significantly reduced the expression of miR-126-5p ( P<0.05). The results of flow cytometry showed that compared with the high glucose model group, the apoptosis rate of the si-HOTAIR group significantly decreased ( P<0.05), while the apoptosis rate of the si-HOTAIR+ miR-126-5p inhibitor group significantly increased ( P<0.05). The CCK-8 experiment showed that compared with the high glucose model group, the cell viability of the si-HOTAIR group significantly increased ( P<0.05); The cell viability of the si-HOTAIR+ miR-126-5p inhibitor group was inhibited ( P<0.05). Conclusions:miR-126-5p can inhibit high glucose induced apoptosis in HK-2 cells and protect them.

BACKGROUND: Peritendinous adhesion that simultaneous with tendon healing link the healing tendon to the surrounding tissue. It results in functional disability, and has a significant adverse impact on health as well as social and economic development. METHODS: Based on a search in the PubMed and Web of Science database, the research articles were screened by their time, main idea, impact factor index, while the ones with no credibility were excluded. Afterwards, we go through the analysis of the reliability and characteristics of the results were further screened from selected articles. RESULTS: A total of 17 biomaterials used to evaluate the adhesion mechanism and the properties of the material were found. All of these biomaterials contained randomized controlled studies and detailed descriptions of surgical treatment that support the reliability of their results which indicates that biomaterials act as barriers to prevent the formation of adhesion, and most of them exhibit satisfactory biocompatibility, biodegradability or selective permeability. Moreover, a few had certain mechanical strength, anti-inflammatory, or carrier capacities. However, there still existed some defects, such as time, technology, clinical trials, material targeting and different measurement standards which also lowered the reliability of their results. CONCLUSION In future, anti-adhesion biomaterials should focus on affordable raw materials with wide sources, and the production process should be simplified, in this way, the versatility and targeting of materials will be improved.

BACKGROUND: Peritendinous adhesion that simultaneous with tendon healing link the healing tendon to the surrounding tissue. It results in functional disability, and has a significant adverse impact on health as well as social and economic development. METHODS: Based on a search in the PubMed and Web of Science database, the research articles were screened by their time, main idea, impact factor index, while the ones with no credibility were excluded. Afterwards, we go through the analysis of the reliability and characteristics of the results were further screened from selected articles. RESULTS: A total of 17 biomaterials used to evaluate the adhesion mechanism and the properties of the material were found. All of these biomaterials contained randomized controlled studies and detailed descriptions of surgical treatment that support the reliability of their results which indicates that biomaterials act as barriers to prevent the formation of adhesion, and most of them exhibit satisfactory biocompatibility, biodegradability or selective permeability. Moreover, a few had certain mechanical strength, anti-inflammatory, or carrier capacities. However, there still existed some defects, such as time, technology, clinical trials, material targeting and different measurement standards which also lowered the reliability of their results. CONCLUSION In future, anti-adhesion biomaterials should focus on affordable raw materials with wide sources, and the production process should be simplified, in this way, the versatility and targeting of materials will be improved.

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.

OBJECTIVE: To investigate the prevalence and relevant influencing factors of gynecological diseases of grassroot level female medical staffs. METHODS: A total of 2 308 female medical workers from county, town and village in hengxian County of Guangxi Zhuang Autonomous Region were selected as study subjects by cluster sampling method. The basic information, occupational history, reproductive health and fertility of these subjects were investigated by Reproducetive Health Survey Questionnaine of Female Workers. RESULTS: The total prevalence of gynecological diseases in female medical staffs was 21.6%. Among them, the prevalence of genital tract infections was 15.6%, and gynecological tumors was 6.0%. The top three gynecologic diseases were vaginitis(9.2%), uterine fibroids(4.3%) and cervicitis(3.5%). Multivariate logistic regression analysis results indicated that the younger the patients, the higher the risk of reproductive tract infectious diseases(P<0.01), and the lower the risk of gynecological tumors(P<0.01). The risk of reproductive tract infectious diseases and gynecological tumors in married patients was higher than that in unmarried staffs(P<0.05). The higher the number of abortions, the higher the risk of reproductive tract infectious diseases and gynecological tumors(P<0.01). The risk of reproductive tract infectious diseases was higher and the risk of gynecological tumors was lower in female shift workers than that of non-shift workers(P<0.05). CONCLUSION: Vaginitis, uterine fibroids and cervicitis are the main gynecological diseases in grassroot level female medical staffs. The incidence of gynecological diseases is related to age, history of marriage, childbirth and abortion, and work-shifts.

OBJECTIVE： To provide reference for perfecting and improving the quality standard of Qiju dihuang oral liquid. METHODS： According to No. 0502 method stated in 2015 edition of Chinese Pharmacopeia （part Ⅳ）， TLC method was used to identify the fruit of Chinese wolfberry， Dendranthema morifolium and Paeonia suffruticosa in Qiju dihuang oral liquid. Using the fruit of C. wolfberry， D. morifolium and paeonol as control， the deployment systems were trichloromethane-ethyl acetate-formic acid （6 ∶ 1 ∶ 0.5， V/V/V）， trichloromethane-isopropanol-formic acid （10 ∶ 1 ∶ 0.5， V/V/V） and cyclohexane-ethyl acetate （3 ∶ 1， V/V）. The contents of morroniside， loganin and paeonol in Qiju dihuang oral liquid were determined by HPLC. The determination was performed on InertSustain C18 column with mobile phase consisted of acetonitrile-0.03％ phosphoric acid solution（gradient elution）at the flow rate of 1.0 mL/min. The detection wavelength was set at 240 nm （morroniside and loganin） and 274 nm （paeonol）， and the column temperature was 40 ℃. The sample size was 10 μL. RESULES： In TLC of the fruit of C. wolfberry， D. morifolium and P. suffruticosa， same color spots were shown in the corresponding positions of reference substance/control chromatogram without interference from negative control. The linear ranges of morroniside， loganin and paeonol were 2.12-106.17， 1.91-95.63 and 4.78-239.16 μg/mL （R2＝0.999 9， 0.999 9， 0.999 8）， respectively. The limits of quantitation were 2.12， 1.91， 2.39 μg/mL； the limits of detection were 0.53， 0.48， 0.59 μg/mL， respectively； RSDs of precision， reproducibility and stability tests were lower than 2％ （n＝6）. The average recoveries were 98.27％， 97.06％ and 97.65％ RSD were 0.80％， 1.18％ and 1.36％ （n＝6）. RSDs of durability tests were all lower than 2％ （n＝3）. CONCLUSIONS： The established method is simple， specific and durable， and can provide reference for improving the quality standard of Qiju dihuang oral liquid.

Objective To explore the characteristics of respiratory parameters in patients with different body mass index during general anesthesia with tracheal intubation. Methods 102 patients scheduled for otitis me-dia surgery were divided into low weight group(B1,n=32),normal weight group(B2,n=36)and overweight or obese group(B3,n = 34 ). After general anesthesia with tracheal intubation,the tidal volume of anesthetic ma-chine wasadjusted to maintain the end tidal carbon dioxide partial pressure between 35 - 45 mmHg. At 10 min (T1),30min(T2)and 60 min(T3)after adjustment,arterial PH,arterial partial pressure of oxygen(PaO2),arte-rial carbon dioxide pressure(PaCO2),inspiratory tidal volume(VTi),expiratory tidal volume(VTe),end tidal carbon dioxide partial pressure(PETCO2),peak airway pressure(Ppeak),plateau airway pressure(Pplat)and dy-namic lung compliance(Cdyn)were recorded. Results PH and PaO2 were not significantly different at T1-3 among the three groups(P>0.05). As compared with group B1 and B2,PaCO2 was lower in group B3. In comparison with group B2,VTi,VTe and Cdyn were higher in group B1 and lower in group B3(P < 0.05). Ppeak and Pplat were lower in group B1 but higher in group B3(P<0.05). PETCO2 was higher in group B1(P>0.05)while lower in group B3 (P < 0.05). Conclusions With the increase in BMI during general anesthesia with tracheal intubation ,the VTi,VTe,Cdyn,PETCO2 and PaCO2 decrease significantly,but Ppeak and Pplat elevate markedly. BMI is a refer-ence index for setting respiratory parameters.

BACKGROUND: It has been believed mesenchymal stem cells (MSCs) play a role in treatment through paracrine mechanism. Various side effects such as embolism, tumorigenesis and immunological reaction caused by direct injection of MSCs can be avoided by extracting MSC lysate. However, there is a larger difference in current collection methods and standards of MSC lysate. OBJECTIVE: To compare repeated freeze-thaw and ultrasonication for the collection of lysate of MSCs. METHODS: Adipose-derived mesenchymal stem cells (ADMSCs) were isolated from the abdominal subcutaneous fat of healthy individuals, and purified with adherence screening method, followed by in vitro amplification using fetal bovine serum medium. The common surface makers of these cells were tested by flow cytometry (1×109, 2×109, 4×109/L). Repeated freeze-thaw and ultrasonication were employed for cell cytoclasis at three different densities respectively in saline and double distilled water, and a comprehensive comparison was performed on cytoclasis rate and the content of protein in cell lysate between the two methods. RESULTS AND CONCLUSION: (1) ADMSCs obtained from in vitro isolated human adipose tissue grew in a swirl or radial pattern with a homogenous size and neat arrangement. CD44, CD90, CD105 and other commonly used surface markers were highly expressed. (2) The study for optimization of lysate collection revealed that the higher cell density implicated a longer time for cell wall disruption and cytoclasis, as well as significantly increased cytoclasis rate. (3) BCA protein assay showed that the highest content of protein was obtained in saline solvent using ultrasonication method. Comprehensive analysis on the results leads to a conclusion that ultrasonication method with saline as the solvent is the optimized method for extraction of ADMSCs lysate, and the cell concentration of less than 4×109/L is recommended.

Sufficient embryos are needed for the preservation of Beagle dogs germplasm resources and the prepara-tion of gene?modified human disease animal models. Up to now, the induced ovulation technique has no effect on dogs,it is hard to obtain mature oocytes in vivo, although the scientists try a lot in many aspects, but still could not make a break?through. The in vitro maturation rate is too low to support the preservation of germplasm resources, application in gene?modified disease models and biomedical research. Aiming to provide useful information on breakthrough in dog oocytes mat?uration, this review will summarize the effect of different age and reproductive stage,different morphology and size of the oo?cytes and lipid droplet on the in vitro maturation of dog oocytes.