Objective The purpose of this study is to construct eukaryotic gene vector of herpes simplex virus type 1 thymidine kinase(HSV1-tk)and to observe the expression of HSV1-tk in lung adenocarcinoma AGZY cell line.Methods The full length HSV1-tk gene was amplified by PCR from plasmid pHSV 106 and was inserted into pMD18-T.The recombinant plasmid was recombined with eukaryotic vector plRES 2-EGFP u-sing gene recombinant technique .HSV1 -tk was transfected into adenocarcinoma AGZY cell line with Lipo-fectamineTM 2 000.Fluorescence microscopy was used to detect the transfection and expression of HSV 1-tk.RT-PCR was used to detect the mRNA levels of HSV 1-tk.The cell proliferation was measured by MTT assay .Re-sults A length of 1 130 bp gene sequence was obtained by PCR .The expressions of HSV 1-tk at mRNA and protein levels were displayed by RT -PCR and Western blot .MTT analysis showed that there were no significant changes cell survival on after transfection .Conclusion The eukaryotic expression vector of HSV 1 -tk report gene is successfully constructed and HSV 1-tk is effectively expressed in transfected AGZY cells .

Objective To observe the injury of some chief organ induced by paraquat (PQ) poisoning in rats, and to explore the mechanism. Method A total of 60 inbred line SD rats were randomly divided into experimental group (n = 30) and control group (n = 30), and each group was further divided into 6 subgroups (n =5) as per the sacrifice of rats at different intervals. The rats of experimental group received the intra-abdominal injection of paraquat (1 mg/mL, 18 mg/kg), and the rats of control group were treated with the same amount of saline solution instead. The rats of each subgroup were sacrificed separately 2 h,6 h, 12 h,24 h,72 h and 120 h after administration of PQ or saline. Lungs, livers and kidneys were taken for histopathological study. Results There was noticeable exudate in lung tissue of rats in experimental group in the early stage. And then the cystic changes in the liver of rats in experimental group were found. A noticeable hemoglobin was found in the renal tubules 24 h after modeling. But the exudation in lung decreased 24 h later, and in the mean time, the disorganization of pulmonary alveoli was obvious and some remarkable collagen appeared in the interstitial tissue of lung, and it was significantly obvious 72 h after modeling. In the liver of rats in experimental group, the injured tissue had some extent of repair in 72 h after modeling, and recovered gradually. But the injury of kidney was exacerbated 72 h after modeling. In the control group, the lung, liver and kidney were not changed in all stages after modeling.Conclusions The paraquat could induce failure of some chief organs in SD rats. The injury was most remarkable in the lung in a progressive way. The kidney injury was not more severe than that of lung tissue, but the pathological changes of the kidney became worse and worse as time taken. The injury of liver induced by paraquat was slight, and the injury could heal up gradually.

OBJECTIVE To investigate the drug resistance of ESBLs-producing Escherichia coli and Klebsiella pneumoniae in local nosocomial infection,for guiding the clinical drug resistance. METHODS ATB analysis system was used for identification of bacteria,extra-susceptibility tests were detected by K-B method. RESULTS The isolation rate of ESBLs-producing E. coli and the K. pneumoniae was 29.9% and 30.8%,respectively. The drug susceptibility was indicated the resistance rate of ESBLs producing strains to antibacterial agents except imipenem was higher than that of non-ESBLs producing strains. CONCLUSIONS Detecting drug resistance of ESBLs producing strains is of important significance for guiding the clinical rational use of antibacterials and controling the epidemics.

Objective To observe the influence of the short effect antihypertension drugs- nifedipine and medial effect antihypertension drugs- extended release nifedipine on the blood pressure variability (BPV) in essential hypertension(EH). Methods Twenty-five EH patients were underwent 24-hour noninvasive ambulatory blood pressure monitoring (ABPM) and observed their BPV respectively before taking drugs, after taking nifedipine and extended release nifedipine. Meantime,25 normotensive controls (NC) were observed. Results (1)BPV in EH group was higher than that in controlled group and the severer the rise of blood pressure, the more obvious the increase of BPV (P 0.05). Conclusions Nifedipine could increase BPV but extended release nifedipine did not change BPV while they decreased blood pressure. Effect of extended release nifedipine was better than nifedipine in decreasing blood pressure.

Objective:To study the changes of immunoglobulins(IgG,IgA,IgM)and helper T cell Th9 subset,and their cor-relation with insulin-like growth factor-1(IGF-1)and S-100B protein in patients with Parkinson's disease(PD).Methods:A total of 108 patients with PD confirmed in Hebei Hospital of Traditional Chinese Medicine were enrolled as study group between December 2020 and December 2022.According to disease severity,they were divided into mild group(35 cases),moderate group(44 cases)and severe group(29 cases);a total of 108 healthy adults were enrolled as control group.Levels of immunoglobulins and Th9 subset were compared between study group and control group,and levels of immunoglobulins,Th9 subset,IGF-1 and S-100B protein were compared among mild group,moderate group and severe group.Correlation between immunoglobulins,Th9 subset and IGF-1,S-100B proteins were analyzed by Pearson correlation analysis.Correlation between disease severity and all differential indexes in PD patients were analyzed by Spearman correlation analysis.Results:IgM level in study group was lower than that in control group,and which was the lowest in severe group,followed by moderate group and mild group(P<0.05).Levels of IgG,IgA,IL-9 and Th9 subset in study group were higher than those in control group,and which were the highest in severe group,followed by moderate group and mild group(P<0.05).IGF-1 level was the lowest in severe group,followed by moderate group and mild group,while level of S-100B protein was on the contrary(P<0.05).Results of Pearson correlation analysis showed that IgM level was positively correlated with IGF-1 level,while negatively correlated with level of S-100B protein.Levels of IgG,IgA,IL-9 and Th9 subset were negatively correlated with IGF-1 level,while positively correlated with level of S-100B protein(P<0.05).Results of Spearman correlation analysis showed that the disease severity was negatively correlated with IgM and IGF-1 levels,while positively correlated with levels of S-100B protein,IgG,IgA,IL-9 and Th9 subset(P<0.05).Conclusion:IgM level is decreased,while levels of IgG,IgA and Th9 subset are increased in PD patients.Changes of the above indexes are significantly correlated with IGF-1 and S-100B,which can be applied to evaluate dis-ease severity.

In order to enhance the antitumor efficacy of recombinant Newcastle disease virus, rNDV-IL15 was rescued in this study. Recombinant plasmid prNDV-IL15 was constructed, and BHK21 cells were transfected with the recombinant plasmid. Finally, the recombinant Newcastle disease virus rNDV-IL15 was successfully rescued. The growth curves of these two recombinant viruses were determined. Murine melanoma B16F10 cells were infected with rNDV-IL15 at MOI of 0.1, and the expression level of IL15 in the supernatant was detected by ELISA. The antitumor efficacy of rNDV-IL15 and rNDV was compared in vitro and in vivo. Results showed that prNDV-IL15 was constructed and recombinant virus rNDV-IL15 was successfully rescued. The growth curve of rNDV-IL15 showed that the growth of rNDV-IL15 had not been changed after insertion of IL15 gene. Results showed that there was high level of IL15 expression in the supernatant of rNDV-IL5-infected B16F10 cells (1 044.3 +/- 27.7 ng x mL(-1)). rNDV-IL15 and rNDV significantly inhibited the growth of B16F10 cells in vitro in a time-dependent manner. However, there was no significant difference between them. In animal experiments, rNDV-IL15 efficiently suppressed tumor growth in vivo when compared with rNDV, and the difference was statistically significant. The results suggested that rNDV-IL15 is a more effective antitumor agent.
Animals ; Body Weight ; Cell Line, Tumor ; Cell Proliferation ; Chick Embryo ; Cytotoxicity, Immunologic ; Female ; Genetic Therapy ; Interleukin-15 ; genetics ; metabolism ; Melanoma, Experimental ; pathology ; therapy ; Mice ; Neoplasm Transplantation ; Newcastle disease virus ; genetics ; Plasmids ; Recombinant Proteins ; genetics ; metabolism ; Transfection ; Tumor Burden

OBJECTIVETo screen for esophageal squamous cell carcinoma (ESCC)-associated long non-coding RNAs (lncRNA) and identify oncogenic lncRNA contributing to ESCC pathogenesis.

METHODSA lncRNA array containing 7419 lncRNA was used to detect the transcriptional profiles of lncRNA of four pairs of ESCC and matched normal esophageal tissue. Bioinformatic analysis was employed to identify differentially expressed ESCC associated lncRNA (ESCCAL). Quantitative real-time PCR was used to verify selected dysregulated lncRNA on independent ESCC samples.

RESULTSGenome-wide transcriptome profiling (coding and or noncoding RNA transcripts) was able to distinguish ESCC from normal tissue. Among these, bioinformatic analysis has identified 154 differentially expressed ESCC associated lncRNA (ESCCALs), which included 111 downregulated and 43 upregulated lncRNA in ESCC relative to the normal tissue (P< 0.01). The highest upregulated lncRNA (ESCCAL_1) and known onco-lncRNA HOTAIR was further verified in 26 paired ESCC samples. ESCCAL_1 and HOTAIR were found to be highly expressed in 17 ESCC and 18 ESCC compared with normal esophageal tissues.

CONCLUSIONThis investigation has revealed large scale aberrant expression of lncRNA in ESCC. About 70% of novel lncRNA-ESCCAL_1, together with a known lncRNA-HOTAIR, are highly expressed in ESSC, suggesting that ESCCAL_1 and HOTAIR may participate in the pathological process of ESCC. Furthermore, lncRNA could be potential diagnostic and prognostic biomarkers for ESCC.

Carcinoma, Squamous Cell ; diagnosis ; genetics ; Esophageal Neoplasms ; diagnosis ; genetics ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Genome-Wide Association Study ; methods ; Humans ; Oligonucleotide Array Sequence Analysis ; Prognosis ; RNA, Long Noncoding ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

Here,we reported a case of term infant with unilateral congenital microphthalmia (CM).Physical examination conducted on 2 d after birth showed that the left eye was barely open and the eye socket was deeper than the right.Meanwhile,the cornea of the left eye was not completely exposed and the light reflex could not be elicited.Ophthalmology consultation,ultrasound and CT scan were conducted,and CM was finally diagnosed since the infant was found to have a small eyeball and shortened axial length,accompanying by pathological changes in the lens,vitreous body and retina.This case suggested that detailed physical examination should be carried out,especially for the eyes,including orbit and eyelid,presence or absence of secretion or concealed eyeball,together with medical imaging techniques technology,to ensure an early detection and diagnosis of CM in neonates.

AIM:To observe the effect of Yiqi Huayu Huatan decoction(YHHD)on unilaterral ureteral ob-struction(UUO)-induced renal interstitial fibrosis in rats, and to investigate the possible mechanism.METHODS: Fe-male SD rats(n=48)were randomly divided into sham group, model group, telmisartan group, and low-, middle-and high-dose YHHD groups,with 8 rats in each group.The UUO model rats was established by ligating left ureter.The rats in sham group and model group were treated with equal volume of normal saline, others were treated with the corresponding drugs daily.After 12 weeks,the rats were sacrificed.The serum samples were collected for determining the concentrations of cystatin C(Cys-C)and uric acid(UA).The morphological changes of the renal tissue were observed by PAS staining. The collagen fiber was observed by Masson staining.The mRNA expression of Krüppel-like factor 15(KLF15),high-mo-bility group box protein 1(HMGB1),nuclear factor-κB(NF-κB),IκB,monocyte chemotactic protein-1(MCP-1),inter-leukin-1β(IL-1β), tumor necrosis factor-α(TNF-α),fibronectin(FN),collagen type I(Col I)and Col-Ⅳwas detec-ted by real-time PCR.The protein expression of KLF15, HMGB1 and NF-κB was detected by Western blot.The protein expression of MCP-1 was determined by the method of immunohistochemistry.RESULTS:Compared with sham group,the deposition rate of collagen fibers and the concentration of Cys-C in model group were significantly increased(P<0.05), the mRNA and protein expression of KLF15 was significantly down-regulated(P<0.05), while the mRNA expression of HMGB1,NF-κB,IκB,MCP-1,IL-1β,TNF-α,FN,Col I and Col Ⅳand the protein expression of HMGB1,NF-κB and MCP-1 were significantly up-regulated(P<0.05).Compared with model group,the deposition rates of collagen fibers in middle-and high-dose YHHD groups and telmisartan group were significantly decreased(P<0.05),with down-regulated protein expression of HMGB1 and NF-κB and mRNA expression of IL-1βand TNF-α(P<0.05).The protein expression of KLF15 was significantly up-regulated in high-dose YHHD group and telmisartan group(P<0.05),while the protein ex-pression of MCP-1 and the mRNA expression of FN were significantly down-regulated(P<0.05).The mRNA expression of KLF15 was significantly up-regulated in high-dose YHHD group(P<0.05), while the mRNA expression of MCP-1, Col I and Col IV was significantly down-regulated(P<0.05).The mRNA expression of NF-κB and IκB was significantly down-regulated and the concentration of Cys-C was significantly decreased in each dose of YHHD groups and telmisartan group(P<0.05).No significant difference of UA level among the groups was observed.CONCLUSION:YHHD allevi-ates renal interstitial fibrosis in a dose-dependent manner, and YHHD at high dose shows the most obvious effect.The mechanism may be associated with the up-regulation of KLF15 and the down-regulation of HMGB1, NF-κB and its down-stream inflammation-related factors in the renal tissue.

Objective:To do genetic analysis on the VP1 gene of 2 Coxsackievirus A12 (CV-A12) isolated from hand, foot and mouth disease (HFMD) surveillance in Yunnan province.Methods:Coxsackievirus isolation was carried out in RD cells from the clinical samples. CV-A12 strains were identified by realtime RT-PCR and sequencing technology from the positive cultures. The VP1 region of the CV-A12 strain was further sequenced. The VP1 gene sequences were initially aligned and then used to construct the phylogenetic tree with GenBank reference strains.Results:The two CV-A12 strains had the highest homology with the Yunnan reference strains in 2018 and the amino acids in VP1 region had specific mutations with other cluster reference strains at multiple sites.Conclusions:The CV-A12 in the HFMD cases in Yunnan province has occured regional specific mutation in VP1 gene.