Objective To investigate effects of α-crystallin on proliferation of lipopolysaccharide (LPS)-activated retinal microglia and bioactivity of iNOS.Methods The retinal microglial cells cultured in vitro were analyzed and their purity was identified by cell immunofluorescence and flow cytometry.After microglia cells being intervened using LPS and α-crystallin at various concentrations,influence of α-crystallin on activity of LPS-activated retinal microglia was detected by MTT method and level of NO was measured by RT-PCR to observe changes of iNOS expression in microglia.Results Purity of primary cultured microglial cells was 94.15％ by GSA-IB4 immunohistochemical identification and 93.34％ by CD11b flow cytometry.α-crystallin of 10-4g/L awakened activity of microglia induced by 10-6g/L LPS (P ＜ 0.01).Expressions of iNOS protein and mRNA showed significant decrease in combined treatment group (P ＜ 0.05).Conclusion In clinical condition,α-crystallin decreases the harm of microglial cells on retinal ganglial cells (RGCs) after optical nerve injury by inhibiting the microglia cells to produce NO and iNOS.

Objective: To know the pre-treatment drug resistance ( PDR ) status of newly reported human immunodeficiency virus type 1 ( HIV-1 ) infected individuals in Wenzhou, so as to provide guidance for antiretroviral therapy ( ART ). Methods: Totally 232 plasma samples of newly reported HIV-1 infected individuals who had not received ART were collected in Wenzhou in 2019. Virus ( HIV-1 ) RNA was extracted, followed by reverse transcription PCR and nested PCR to amplify the pol region and sequence. Resistance mutations and resistance to non-nucleoside reverse transcriptase inhibitors ( NNRTIs ), nucleoside reverse transcriptase inhibitors ( NRTIs ) and protease inhibitors ( PIs ) was analyzed. Results: The pol region sequences from 199 infected patients were obtained and the incidence of PDR was 8.04% ( 16/199 ). Eight genotypes were detected, including circulating recombinant forms ( CRFs ) CRF07_BC ( 47.24%, 94/199 ) and CRF01_AE ( 29.15%, 58/199 ) which were the dominant types. Two unique recombinant forms ( URFs ) were detected, namely URF( CRF01_AE/BC ) and URF( B/C ) . Thirty-one cases ( 15.58% 31/199 ) had drug-resistant mutations. For NNRTIs, NRTIs and PIs, 20 cases ( 64.52% ) , 2 cases ( 6.45% ) and 9 cases ( 29.03% ) with drug resistance mutations were detected, respectively. The resistance mutations to NNRTIs included K101E, K103N/R, V106I, E138K, V179D/E/T, Y181C, G190A and H221Y. Four cases each had two resistance mutations to NNRTIs. The resistance mutations to NRTIs were V75M and M184V. The resistance mutations to PIs were M46I, L33F and Q58E. For the newly released NNRTI drug Doravirine ( DOR ), two cases were found to have mutations of resistance. Conclusions The incidence of PDR among newly reported HIV-1 patients in Wenzhou is 8.04%, mainly caused by NNRTIs drug-resistant mutation. Resistance to the new drug DOR has emerged. The surveillance of drug resistance should continue to be strengthened.

Objective To investigate the expression of miR-155 in rats exposed to PM2.5 and its signifi-cance.Methods Thirty-two 12-weeks-old healthy male SD rats were randomly divided into PM2.5-primary expo-sure group (P1),PM2.5-three times exposure group (P3), saline-control group and blank-control group (O), with 8 rats in each group. The pathological changes of lung tissue of each group were observed by HE staining and the expressions of miR-155 in lung tissue of each group were measured by RT-PCR. The expressions of TNF-α, IL-6 and IL-1β in serum of each group were measured by ELISA.One-way ANOVA and LSD-t test were used for statistical analysis,and Pearson′s correlation analysis was used to evaluate the relationship between the levels of miR-155 and TNF-α,IL-6 and IL-1β. Results The expression of inflammatory cytokines TNF-α,IL-6 and IL-1β in P1 and P3 increased when compared with that in saline control group and blank-control group, and the amount of expression increased with the increase of the number of exposure and dose;the difference between groups was statistically sig-nificant(P<0.01).At the same time,with the increase of the dose and number of PM2.5,the inflammatory damage of lung tissue was enhanced.The expression of miR-155 in lung tissue also increased with the increase of the dose and number of PM 2.5,and it had significantly positive correlation with serum inflammatory cytokines(P<0.01),and had statistical significance. Conclusions The lung tissue of rats exposed to PM2.5 can produce inflammatory injury, and the damage is enhanced with the increase of exposure times and dose.The expression of miR-155 is positively correlated with inflammatory injury in rats.Consequently,miR-155 may participate in PM2.5-induced inflammatory lung injury in rats.

Objective:To construct and validate a risk prediction model for immune checkpoint inhibitor-associated pneumonia (CIP) using machine learning algorithms and the nomogram, aiming to provide an accurate and intuitive method to assist nurses in screening people at high risk of developing CIP.Methods:This was a retrospective case -control study. A total of 230 oncology patients treated with immune checkpoint inhibitors attending Zhujiang Hospital of Southern Medical University from January 2019 to February 2022 were collected using the hospital's electronic medical record system. The prediction models were built using five machine learning algorithms and nomogram. The models were then validated on a separate test set, and their differentiation and stability were assessed using evaluation indices like AUC and accuracy rate.Results:Underlying lung disease, smoking history, serum albumin≤35 g/L and radiotherapy history were identified as important influencing factors of CIP in all six models. The AUC of K nearest neighbor, support vetor machines (SVM), naive Bayesian, decision tree and random forest models predicted CIP were 0.647, 0.696, 0.930, 0.870, and 0.934, respectively. The AUC of the model created by the nomogram was 0.813, which was lower than the best random forest model in the machine learning algorithm, but with good predictive performance (AUC=0.934).Conclusions:The nomogram model can assess the patient′s risk more intuitively, but the risk prediction model of CIP based on a machine learning algorithm has a higher diagnostic value. It is suggested that the accuracy and usefulness of the prediction model can be increased by combining the nomogram's foundation with the machine learning algorithm.

Objective To understand the occurrence of healthcare-associated infection (HAI),distribution of pathogens,and drug resistance in a general hospital in 2014-2016,provide basis for prevention and control of HAI.Methods Clinical data of hospitalized patients from January 2014 to December 2016 were collected by prospective and retrospective investigation,distribution and drug resistance of pathogens causing HAI were statistically analyzed.Results From 2014 to 2016,4 750 patients had 5 352 cases of HAI,incidence and case incidence of HAI were 2.19％ and 2.46％ respectively.Incidences of HAI in three years were 2.47％,2.07％,and 2.05％ respectively,showing a decreased tendency,difference was statistically significant (x2 =36.217,P＜0.01).Incidences of HAI were high in intensive care unit,department of neurosurgery,as well as department of burn and plastic surgery,the common HAI sites were respiratory tract,urinary tract,and surgical sites.The main pathogens causing HAI were gram-negative bacteria (76.10％).Resistance rates of Escherichia coli to cephalosporins and fluoroquinolones were relatively higher (＞60％);resistance rates of Klebsiella pneumoniae to carbapenems were relatively higher;resistance rates of Pseudomonas aeruginosa to carbapenems showed a increased tendency year by year (x2 =15.175,P =0.001);antimicrobial resistance rates of Acinetobacter baumannii were all＞50 ％.Methicillin-resistant Staphy lococcus aureus (SA) accounted for about 60％ of SA,methicillin-resistant coagulase negative Staphylococcus(CNS) accounted for more than 80％ of CNS,vancomycin-and linezolid-resistant Staphylococcus spp.were not found.Conclusion The common pathogens causing HAI in this hospital are higher.Scientific monitoring on HAI and regular analysis of clinical data are of great significance for guiding rational use of antimicrobial agents,controlling multidrug-resistant organisms,and reducing the occurrence of HAI.

This study investigated the effects of microRNA-135a (miR-135a) targeting of glycogen synthase kinase 3β (GSK3β) on the epithelial–mesenchymal transition (EMT), migration and invasion of bladder cancer (BC) cells by mediating the Wnt/β-catenin signaling pathway. BC and adjacent normal tissues were collected from 165 BC patients. Western blotting and quantitative real-time PCR were used to detect the expression of GSK3β, β-catenin, cyclinD1, E-cadherin, vimentin and miR-135a in BC tissues and cells. Cells were assigned to blank, negative control (NC), miR-135a mimics, miR-135a inhibitors, small interfering RNA (siRNA)-GSK3β or miR-135a inhibitors+siRNA-GSK3β groups. miR-135a, β-catenin, cyclinD1 and vimentin expression increased, while GSK3β and E-cadherin expression decreased in BC tissues compared with adjacent normal tissues. Compared with the blank and NC groups, the expression of miR-135a, β-catenin, cyclinD1 and vimentin was higher, and cell proliferation, migration, invasion and tumor growth were increased in the miR-135a mimics and siRNA-GSK3β groups. These groups showed an opposite trend in GSK3β and E-cadherin expression and cell apoptosis. The miR-135a inhibitors group was inversely correlated with the blank and NC groups. It was concluded that miR-135a accelerates the EMT, invasion and migration of BC cells by activating the Wnt/β-catenin signaling pathway through the downregulation of GSK3β expression.
Apoptosis ; Blotting, Western ; Cadherins ; Cell Proliferation ; Down-Regulation ; Glycogen Synthase Kinases ; Humans ; Negotiating ; Real-Time Polymerase Chain Reaction ; RNA, Small Interfering ; Urinary Bladder Neoplasms ; Urinary Bladder ; Vimentin

OBJECTIVETo investigate the relationship of sperm morphology with reproductive hormones in infertile men and the pathogenesis of teratozoospermia.

METHODSThis study included 90 infertile men aged 25 - 40 years. We measured their testis volumes using the Prader orchidometer, conducted routine semen analyses according to the WHO laboratory standard, and determined the concentrations of reproductive hormones and sex hormone-binding globulin (SHBG) by chemiluminescence and the levels of free testosterone (FT) and bioavailable testosterone (BioT).

RESULTSAll the subjects showed normal sperm concentration. Based on the results of semen morphology analysis, the 90 infertile men were equally divided into groups 1 (morphologically normal sperm <4%), 2 (morphologically normal sperm > or = 4% and <10%), and 3 (morphologically normal sperm > or = 10%), with no significant differences in age among the three groups (P>0.05). The volumes of the left testis were (14.27 +/- 3.65) ml, (16.90 +/- 3.57) ml and (14.57 +/- 3.57) ml, respectively (P = 0.006 group 1 vs group 2, P = 0.741 group 1 vs group 3, P = 0.014 group 2 vs group 3), and those of the right testis were (14.60 +/- 3.70) ml, (16.60 +/- 3.35) ml and (14.67 +/- 3.54) ml, respectively (P = 0.050). There were no significant differences among the three groups in prolactin, follicle-stimulating hormone, luteinising hormone, estradiol, total testosterone and SHBG, (P>0.05). The levels of serum FT were (0.25 +/- 0.07) nmol/L, (0.29 +/- 0.07) nmol/L and (0.31 +/- 0.13) nmol/L (P = 0.086 group 1 vs group 2, P= 0.010 group 1 vs group 3, P= 0.364 group 2 vs group 3), and those of BioT were (5.81 +/- 1.58) nmol/L, (6.78 +/- 1.55) nmol/L and (7.29 +/- 3.02) nmol/L, respectively (P = 0.086 group 1 vs group 2, P = 0.010 group 1 vs group 3, P = 0.364 group 2 vs group 3). The percentage of morphologically normal sperm was positively correlated with the levels of serum FT and BioT (P<0.05).

CONCLUSIONThe higher the levels of serum FT and BioT, the higher the percentage of morphologically normal sperm, which suggests that serum FT and BioT might be involved in the pathogenesis of teratozoospermia.

Adult ; Estradiol ; blood ; Follicle Stimulating Hormone ; blood ; Humans ; Infertility, Male ; blood ; physiopathology ; Luteinizing Hormone ; blood ; Male ; Prolactin ; blood ; Semen ; Semen Analysis ; Sex Hormone-Binding Globulin ; metabolism ; Sperm Count ; Spermatozoa ; abnormalities ; Testis ; Testosterone ; blood

This study was aimed to investigate the reversal effect of 5-bromotetrandrine (5-BrTet) and magnetic nanoparticle of Fe(3)O(4) (Fe(3)O(4)-MNPs) combined with DNR in vivo. The xenograft leukemia model with stable multiple drug resistance in nude mice was established. The two sub-clones of K562 and K562/A02 cells were respectively inoculated subcutaneously into back of athymic nude mice (1 x 10(7) cells/each) to establish the leukemia xenograft models. Drug resistant and the sensitive tumor-bearing nude mice were both assigned randomly into 5 groups: group A was treated with NS; group B was treated with DNR; group C was treated with nanoparticle of Fe(3)O(4) combined with DNR; group D was treated with 5-BrTet combined with DNR; group E was treated with 5-bromotetrandrine and magnetic nanoparticle of Fe(3)O(4) combined with DNR. The incidence of tumor formation, growth characteristics, weight and volume of tumor were observed. The histopathologic examination of tumors and organs were carried out. The protein levels of BCL-2, BAX, and Caspase-3 in resistant tumors were detected by Western blot. The results indicated that 5-BrTet and magnetic nanoparticle of Fe(3)O(4) combined with DNR significantly suppressed growth of K562/A02 cell xenograft tumor, histopathologic examination of tumors showed the tumors necrosis obviously. Application of 5-BrTet and magnetic nanoparticle of Fe(3)O(4) inhibited the expression of BCL-2 protein and up-regulated the expression of BAX, and Caspase-3 protein in K562/A02 cell xenograft tumor. It is concluded that 5-bromotetrandrine and magnetic nanoparticle of Fe(3)O(4) combined with DNR have significant tumor-suppressing effect on MDR leukemia cell xenograft model.
Animals ; Antineoplastic Agents ; pharmacology ; Benzylisoquinolines ; pharmacology ; Daunorubicin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Drug Synergism ; Female ; Ferric Compounds ; administration & dosage ; Humans ; K562 Cells ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Nanoparticles ; administration & dosage ; Xenograft Model Antitumor Assays

This study was purposed to investigate the effects of magnetic nanoparticle of Fe3O4 (Fe3O4-MNPs) on murine immune system. ICR mice were assigned randomly into four groups which were treated with normal saline, low, middle and high dose of MNP-Fe3O4 respectively. The mice were killed after being exposed by intragastric administration for 2 weeks. The ratios of spleen weight to body weight, lymphocyte transformation rate in spleen suspension and phagocytic index of macrophage in abdominal cavity were detected. The results showed that the ratios of spleen weight to body weight in Fe3O4-MNP groups were not significantly different in comparison with the control (p > 0.05). The lymphocyte transformation rate in spleen suspension in Fe3O4-MNP groups were all higher than that in control group (-0.1775 +/- 0.0246), especially in the middle dose group (0.1833 +/- 0.0593) (p < 0.05), and the phagocytic index of macrophages in abdominal cavity of middle dose group (0.2051 +/- 0.0213) was higher than that of control group and other two Fe3O4-MNP group (low dose 0.1538 +/- 0.0100, high dose 0.1511 +/- 0.0184) (p < 0.05). It is concluded that suitable dose of Fe3O4-MNP can enhance the cellular immune activity and phagocytic function of macrophages of mice.
Animals ; Immunity, Cellular ; Lymphocytes ; drug effects ; Macrophages ; drug effects ; Magnetite Nanoparticles ; administration & dosage ; Mice ; Mice, Inbred ICR ; Phagocytosis

BACKGROUNDThe multidrug resistance (MDR) associated with the expression of the mdr1 gene and its product P-glycoprotein is a major factor in the prognosis of hepatocellular carcinoma cell (HCC) patients treated with chemotherapy. Our study was to establish a stable HCC MDR cell line where a de novo acquisition of multidrug resistance specifically related to overexpression of a transgenic mdr1.

METHODSThe 4.5-kb mdr1 cDNA obtained from the plasmid pHaMDR1-1 was cloned into the PCI-neo mammalian expression vector, later was transferred by liposome to human hepatocarcinoma cell line HepG2. Then the transfected HepG2 cells resisting G418 were clustered and cultured and the specific fragment of mdr1 cDNA, mRNA and the P-glycoprotein (Pgp) in these HepG2 cells were detected by PCR, RT-PCR and flow cytometry, respectively. The accumulation of the daunorubicin was determinated by flow cytometry simultaneously. The nude mice model of grafting tumour was established by injecting subcutaneously HepG2/mdr1 cells in the right axilla. When the tumour diameter reached 5 mm, adriamycin was injected into peritoneal cavity. The size and growth inhibition of tumour were evaluated.

RESULTSThe mdr1 expression vector was constructed successfully and the MDR HCC line HepG2/mdr1 developed. The PCR analysis showed that the specific fragment of mdr1 cDNA in HepG2/mdr1 cells, but not in the control group HepG2 cells. Furthermore, the content of the specific fragment of mdr1 mRNA and Pgp expression in HepG2/mdr1 cells were (59.7 +/- 7.9)% and (12.28 +/- 2.09)%, respectively, compared with (16.9 +/- 3.2)% and (3.07 +/- 1.06)% in HepG2 cells. In the nude mice HCC model, the tumour genes of both groups were identified. After ADM therapy, the mean size of HepG2 cell tumours was significantly smaller than HepG2/mdr1 cell tumours.

CONCLUSIONThe approach using the transfer of mdr1 cDNA may be applicable to the development of MDR hepatocarcinoma cell line, whose MDR mechanism is known. This would provide the experimental basis of MDR research.

ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Animals ; Carcinoma, Hepatocellular ; drug therapy ; genetics ; pathology ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; therapeutic use ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Female ; Flow Cytometry ; Genetic Vectors ; genetics ; Humans ; Liver Neoplasms, Experimental ; drug therapy ; genetics ; pathology ; Mice ; Mice, Nude ; Mitomycin ; pharmacology ; therapeutic use ; Reverse Transcriptase Polymerase Chain Reaction ; Xenograft Model Antitumor Assays ; methods