Background and purpose:miR-22 has been reported to be down-regulated in gastric cancer, lung cancer, colorectal cancer, and breast cancer. However, its expression in glioma was still poorly known. This study aimed to explicit whether miR-22 suppresses cell proliferation by targeting MTDH, thus to reveal molecular mechanism that miR-22 functions as a tumor suppressor in glioma. Methods:Quantitative real-time polymerase chain reaction (qRT-PCR) was conducted for detecting the expression of miR-22 in gliomas and normal brain tissues. MTDH 3’UTR-luciferase vector was constructed and dual-luciferase reporter gene assay was employed to examine the effect of miR-22 on luciferase activity. U251 cells were transfected with miR-22 mimics, and MTDH siRNA as for postive control, then Western blot was performed to detect the expressions of MTDH protein. The proliferation ability of U251 cells was evaluated by MTT assay. Results:miR-22 was down-regulated in glioma tissues. Glioma patients with relatively high expression of miR-22 showed lower mortality compared with low expression of miR-22 by using Kaplan-Meier survival curves. We demonstrated miR-22 could bind to the 3’ untranslated region (UTR) of MTDH and inhibited the luciferase activity. Western blot showed that the expression of MTDH protein was inhibited by restored miR-22 or siR MTDH in U251 cells. Overexpression of miR-22 or siR MTDH inhibited the proliferation of U251 cells. Conclusion:miR-22 suppresses cell proliferation by targeting MTDH in glioma.

Abstract: Objective　To investigate the fundus arteriosclerosis and its influencing factors in HIV/AIDS patients after long-term highly active antiretroviral therapy (HAART). Methods　The clinical basic data and fundus examination data of 203 HIV/AIDS patients before and after HAART in the Fourth People 's Hospital of Nanning from January 2020 to June 2022 were collected to evaluate the occurrence of fundus arteriosclerosis and analyze its influencing factors. Results　Of the 203 HIV/AIDS patients, 159 patients developed fundus arteriosclerosis, with an incidence of 78.33%, including 33 patients with grade Ⅰ(20.75%), 87 patients with grade Ⅱ (54.72%), 28 patients with grade Ⅲ(17.61%), and 11 patients (6.92%) with Grade Ⅳ. Before HAART, there was no significant difference in CD4+T lymphocyte count, CD8+T lymphocyte count, viral load, white blood cell count, platelet count, hemoglobin, serum creatinine, blood urea nitrogen, triacylglycerol, total cholesterol, fasting blood glucose, alanine aminotransferase, aspartate aminotransferase and serum total bilirubin between the atherosclerosis group and normal group (P>0.05). After 6 months of HAART, CD8+T lymphocyte count, triacylglycerol and fasting blood glucose in atherosclerosis group were significantly higher than those in normal group (P<0.05). In the stratified comparison of CD4+ and CD8+ lymphocyte counts after 6 months of HAART, the proportion of patients with CD4+ lymphocyte count (CD4+)<200 (cells/μL) in the atherosclerosis group was significantly higher than that in the normal group; the proportion of patients with CD4+ lymphocyte count (CD4+)≥500 (cells/μL) was significantly lower than that in the normal group; the proportion of patients with CD8+ lymphocyte count CD8+≥ 800/μL was significantly higher than that in the normal group (all P<0.05). Binary logistic regression analysis showed that opportunistic infection, HIV course, CD4+T, CD8+T lymphocyte count after HAART and triglyceride were independent risk factors for ocular fundus atherosclerosis in HIV/AIDS patients (all P<0.05). Conclusions　The incidence of ocular fundus arteriosclerosis is high in HIV/AIDS patients. More than 4 years of HIV course, combined opportunistic infection, Low CD4+T lymphocyte count after 6 months of HAART, high CD8+T lymphocyte count and high triglyceride level are independent risk factors for ocular fundus arteriosclerosis in HIV/AIDS patients. Fundus screening should be performed before and after HAART in such population, HAART program should be formulated for the risk of cardiovascular disease, and risk management of cardiovascular disease should be strengthened during treatment to improve patient outcomes.

ObjectiveTo study the effects of early rehabilitation on symptom, range of motion (ROM) and motor function of upper extremity of patients with shoulder-hand syndrome after stroke.Methods47 patients were divided into two groups,early rehabilitation group and non-early rehabilitation group.Two groups were treated with comprehensive rehabilitation. The passive ROM, the pain and motor function of affected limb was assessed by Fugl-Meyer Assessment.The evalution was done in pre-treatment and post-treatment respectively. Results1.The passive ROM decreased and the pain of affected limb of the early rehabilitation group were less than the non-early rehabilitation gronp when the shoulder-hand syndrome occour.2.After comprehensive rehabilitation treatment,their clinical symptom ROM and motor function of upper extremity impoved significantly in patients with shoulder-hand syndrome after stroke( P<0.05).The early rehabilitation treatment has better effects. Conclusions The early rehabilitation treatment could decrease the degree of their passive ROM decrease and the pain of affected limb in patients with shoulder-hand syndrome after stroke,and the patients who accepted earlyrehabilitation seemed easly to improve by comprehensive treatment.

Objective To construct cDNA entry library and cDNA expression library of Armillifer agkistrodontis (A.) nymphs and make a preliminary immunoscreening for the cDNA expression library.Methods The nymphs were collected from the Kunming mice infected experimentally with A.agkistrodontis eggs and the total RNA were extracted from the nymphs using TRIzol Reagent.After purifying the mRNA,the synthesized cDNAs were cloned into the donor vector pDONR222 by BP reaction of Gateway technology and the recombinants were transformed into the DH10B cells by electroporation,the cDNA entry library was obtained.Next,the expression vector pDEST17 was ligated with entry clones by LR reaction,and the recombinants were transformed into the BL21 (DE3) cells.Hence,the cDNA expression library was constructed.Then,the expression library was immunoscreened with the mixed sera of mice infected with A.agkistrodontis,and the insertions of positive clones were sequenced.After that,the open reading frame(ORF) of positive slone sequence,the homology of the screened genes and their encoded proteins were analyzed by Finder and BLAST (basic local alignment search tool) program of National Center of Biotechnology Information(NCBI),and the discovered new genes were submitted into the GenBank.Besides,the physico-chemical properties,secondary structure and B cell epitopes of encoded proteins were also analyzed by bioinformatics software.Results The average titer and total clones of the cDNA entry library were 1.45 × 105 CFU/ml(colony-forming unit,CFU) and 1.74 × 106 CFU,respectively,and the range of fragment length of the inserted cDNA was between 0.2-4.0 kb,with an average of 1.4 kb.The total clones of cDNA expression library were 1.00 × 105 CFU,and the fragment length of the inserted cDNA was between 0.3-2.2 kb,with an average of 1.0 kb.Five positive clones,coded S1,S5,A1,D1 and F1,respectively,were obtained through preliminary immunoscreening.The sequence and homology of the five positive clones were sequenced and analyzed by BLAST program.No significant similarities were found in pentastomida species,which meant that they were all novel genes of A.agkistrodontis.The gene sequences were submitted to GenBank,with the accession number from JQ180451 to JQ180455.Also,results obtained by bioinformatics software showed that the predictive encoding proteins were all potential to be valuable recombinant diagnostic antigens.Conclusions The cDNA library of A.agkistrodontis nymphs is successfully constructed,and five new genes of A.agkistrodontis are discovered.The establishment of cDNA library and the discovery of the new genes will lay a foundation for further studying the gene functions and screening the immunodiagnostic antigens.

Background Some drugs with inhibitory effect on the proliferation of lens epithelial cells have a limiting application in clinic because of their adverse response.To screen the effective and less side-effect drug for supressing LECs growth is very inportant for the prevention and treatment of after cataract.Objective This study was to explore the effects of docetaxel on LECs growth and compare its role with epirubicin hydrochloride,pirarubicin hydrochloTide and rahitrexed.Methotis Immortalized human LECs line (SRA01/04) were cultured and passaged.Different concentrations of docetaxel,epirubicin hydrochloride,pirarubicin hydrochloride and rahitrexed were added into the medium respectively for 24.48 and 72 hours.The proliferation of LECs was detect by M1Yr.Flow cytometry analysis Was used to analyze the influence of different concentrations of docetaxel on cellular cycle at 48 hours after addition of docetaxel,and Annexin V-FITC/PI marking method was used to assesse the apoptosis of LECs under the action of docetaxel.Expression of bcl-2 protein in LECs Was evaluated by Westeru blot. Result The growth rate of LECs Wag 100%in 8-519 pmol/L doeetaxel groups with the normal cell shape.Majority of abnormal cells and low growth rate were found in 66 nmoVL docetaxel group at 48 and 72 hours.The IC50 of docetaxel was lowest in 48 and 72 hours in docetaxel group in comparison to epirubicin hydrochloride and pirarubicin hydrochloride. However,no evident inhibition on LECs growth in 23.22-523.56 μmol/L of raltitrexed.At 48 hours,the percentage of LECs in G2/M phase increased as the asccnte of concentration of docetaxel,showing a significant difference among 4 groups(F=2633.05,P＜0.01).The percentage of early apoptotic cells increased to 22.4%(χ2=20.00,P＜0.01) and 27.9%(χ2=42.68,P＜0.01)from normal control 3.1% at 48 hours after LECs exposed to 8.3 nmol/L and 266 nmol/L docetaxe.The expression of bcl-2 protein in LECs was obviously weakened after addition of docetaxel,especially 8.3 nmol/L docetaxel group. Conclusion Docetaxel,epirubicin hydrochloride and pirarubicin hydrochloride can inhibit the proliferation of human LECs in vitro.But there is no supression on LECs growth inraltitrexed.Docetaxel is proved to have a strongly arrested effect on the proliferation of LECs in comparison with epirubicin hydrochloride and pirarubicin hydrochloride and play its role at concentration-and time-dependent manner.

Objective:To explore the biological function of miR-132 in ovarian cancer and the target. Methods: 22 cases ovarian cancer tissue and non-tumor tissue adjacent were collected,the expression of miR-132 in tumor tissue and non-tumor tissue, normal ovarian epithelial cells and ovarian cancer cell were detected by RT-PCR. The normal ovarian epithelial cells which the expression of miR-132 maximum or minimum were chosen, and they were divided into two groups, respectively with transfection of negative control plasmid ( NC) and miR-132 mimic plasmid. The expression of miR-132 after transfection was detected by RT-PCR,the cell proliferation and cell apoptosis were detected by CCK-8 method and flow cytometry instrument respectively,the expression of Ezrin protein was detected by Western blot. Results:The expression of miR-132 in tumor tissue was significantly lower than the tumor tissue adjacent,the expression of miR-132 in ovarian cancer cell lines was significantly lower than normal ovarian epithelial cells, the differences were statistically significant (P<0. 05). The SKOV3 cell lines was chosed for gene transfection,compared with NC group, transfection with miR-132 mimic plasmid could significantly reduce cell proliferation, increase cell apoptosis, the difference had statistical significance ( P<0. 05 ) . Western blot results showed that up-regulation miR-132 significantly increased the Ezrin protein expression in ovarian cancer SKOV3 cells ( P<0. 05 ) . Conclusion: In ovarian cancer, miR-132;inhibits proliferation and induces apoptosis of ovarian cancer via Ezrin,it may be a tumor suppressor gene.

Objective To explore the clinical significance of preoperative ultrasound-assisted drawings in location of thyroid micronodule resection .Methods A total of 88 patients ( 137 nodules ) who underwent thyroid micro-nodule resection were enrolled in the prospective randomized controlled study .All patients were randomly divided into two groups:46 patients (68 micronodules) in experimental group and 42 patients (69 micronodules) in control group.Inclusion criteria:the maximum diameter of nodule≤1.0 cm. Preoperative thyroid ultrasound was conducted .Patients in experimental group also underwent ultrasound-assisted location of thyroid micronodule .Results The diagnostic accuracy rate of US was 81.82%(69/88) in all patients .With the help of ultrasound-assisted location , all nodules in experimental group were found quickly and accurately.The resection rate of experimental group was 100%(46/46).Whereas 4 nodules (in 4 patients ) were missed in control group with a resection rate of 90.5% (38/42).The postoperative US examinations after 3 months showed that all nodules were completely removed in experimental group while 4 nodules retained in control group .Conclusions Preoperative ultrasound-assisted drawings in location of thyroid micronodule plays an important role in thyroid nodule resection .It is great value of clinical application .

OBJECTIVEIn this study, we aimed to investigate the expressions of adhesion molecules on human bronchial epithelial cells and neutrophils in co-culture system, assess the effects of puerarin on suppressing these adhesion molecules expressions, and explore the roles of two crucial signal-transduction elements p38 mitogen-activated protein kinase (p38 MAPK) and nuclear factor kappa B (NF-κB) in modulating adhesion molecules expressions.

METHODSNeutrophils and BEAS-2B cells (one human bronchial epithelial cell line) were co-cultured, and adhesion molecules expressions on cell surface were detected using flow cytometry. The mRNA levels of adhesion molecules were assessed by real-time quantitative polymerase chain reaction (real-time qPCR). Phosphorylated p38 MAPK and inhibitor κB were analyzed by Western blot.

RESULTSIn co-culture system, adhesion molecules expressions on BEAS-2B cells and neutrophils were enhanced significantly (P<0.05). Correspondingly, the mRNA levels of adhesion molecules were also increased greatly. Moreover, the pretreatment of peurarin obviously suppressed adhesion molecules expressions on cell surface. Furthermore, phosphorylated p38 MAPK and inhibitor κB in BEAS-2B cells and neutrophils were elevated in co-culture system, but decreased significantly after upon the treatment of peurarin (P<0.05).

CONCLUSIONSCoculture boosted the interactions between human bronchial epithelial cells and neutrophils mimicking airway inflflammation, whereas peurarin decreased the expression of adhesion molecules on cell surface by suppressing the activities of p38 MAPK and NF-κB pathways, and exhibiting its anti-inflflammation activity.

Animals ; Base Sequence ; Bronchi ; cytology ; enzymology ; metabolism ; Cattle ; Cell Adhesion Molecules ; metabolism ; Cell Line ; Coculture Techniques ; DNA Primers ; Down-Regulation ; drug effects ; Epithelial Cells ; enzymology ; metabolism ; Isoflavones ; pharmacology ; NF-kappa B ; metabolism ; Neutrophils ; enzymology ; metabolism ; Phosphorylation ; Real-Time Polymerase Chain Reaction ; p38 Mitogen-Activated Protein Kinases ; metabolism

Rat aorta media, adventitia and cultured vascular smooth muscle cells (VSMCs) were used in this study to identify the source of nitric oxide (NO) generation from various cell types of vascular tissues and to elucidate the mechanisms involved in NO formation. Treatment of vascular media and VSMCs with lipopolysaccharide (LPS) or cytokines [tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1beta)] resulted in a dose-dependent increase of NO release. Inducible nitric oxide synthase (iNOS) in the stimulated VSMCs was significantly upregulated as shown by Western blot analysis. Protein kinase C (PKC) inhibitor 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H7) prevented LPS-, TNF-alpha- and IL-1beta-induced NO production, whereas N-(2-guanidinoethyl)-5-isoquinoline-sulfonamide (HA1004), an H7 analogue with little activity towards PKC, had no inhibition effect. The role of PKC in LPS- and cytokine-induced changes on NO formation was confirmed by using another structurally distinct PKC inhibitor chelerythrine. Treatment of VSMCs with protein tyrosine kinase (PTK) inhibitor genistein or tyrphostin AG18 also reduced the NO production evoked by LPS, TNF-alpha or IL-1beta, which was associated with inhibition of iNOS protein expression. In contrast, PKC inhibitor chelerythrine did not affect iNOS expression. These results suggest that PTK mediates LPS- and cytokine-induced NO formation by upregulation of iNOS expression. PKC may be involved in the post-translational modification of iNOS or the regulation of the availability of iNOS substrates and cofactors.
Animals ; Aorta, Thoracic ; cytology ; Cells, Cultured ; Cytokines ; pharmacology ; Interleukin-1beta ; pharmacology ; Lipopolysaccharides ; pharmacology ; Male ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase Type II ; metabolism ; Protein Kinase C ; physiology ; Protein-Tyrosine Kinases ; physiology ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; pharmacology

Adult ; Anemia, Aplastic ; chemically induced ; genetics ; immunology ; Benzene ; poisoning ; Gene Expression ; Genes, T-Cell Receptor beta ; genetics ; Humans ; Male