Objective To study the clinical features and protective measures of AIDS with fungal esophagitis as the initial presentation,improve the knowledge of non-specialist medical staff of AIDS. Methods The clinical manifestations,laboratory tess.endoscopic examination,and medical protective measures of 15 AIDS patients with fungal esophagitis were analyzed. Reults The patients were infected through sexual transmission mainly,and manifestation included difficuhy in swallowing,retrostemal pain,weight loss,fever,loss of appetite,fatigue and other symptoms.The patients got confirmed diagnosis by our hospital after visiting many other hospitals.3 patients were automatically discharged after confirmed diagnosis,other patients impreved after antifimgal therapy and symptomatic treatment.No iatrogenic infection occurred during hospitalization. Conclusions Non-specialist medical staff in AIDS should recognize the clinical features of AIDS,give timely confirmed diagnosis and treatment,avoid missed diagnosis and misdiagnosis,strengthen protective measures and avoid iatrogenic infection.

Objective: To investigate the survival rate of female breast cancer patients in Panyu District, Guangzhou City, so as to provide the basis for improving the prognosis of breast cancer patients. Methods: Basic information including age, clinical stage and surgical treatment of female breast cancer patients registered in Panyu District and diagnosed in 2017 were collected through the Guangzhou Municipal Cancer Registration and Reporting Management System, and were followed up until December 31, 2022. The survival rate was calculated using the life table. Factors affecting survival time among female breast cancer patients were analyzed using a multivariable Cox proportional hazard regression model. Results: A total of 227 female breast cancer patients were reported in Panyu District in 2017, and had a median age of 51 (interquartile range, 17) years. There were 43 cases (18.94%) in stage Ⅰ, 55 cases (24.23%) in stage Ⅱ, 63 cases (27.75%) in stage Ⅲ, 27 cases (11.89%) in stage Ⅳ, and 39 cases (17.18%) with unknown staging. Surgical treatment was performed in 204 cases (89.87%), and chemotherapy was administered in 73 cases (32.16%). By December 31, 2022, there were 40 deaths and 14 patients lost to follow-up. The one-year, three-year and five-year survival rates were 96.44%, 87.45% and 82.87%, respectively. Multivariable Cox proportional hazard regression analysis showed that older age (HR=1.023, 95%CI: 1.002-1.046), clinical stage Ⅲ (HR=10.050, 95%CI: 1.324-76.270) or IV (HR=42.663, 95%CI: 5.588-325.742) were associated with a higher risk of mortality in female breast cancer patients, while surgical treatment (HR=0.278, 95%CI: 0.130-0.598) was associated with a lower risk of mortality. Conclusions The five-year survival rate of female breast cancer patients in Panyu District was 82.87%. Age, clinical stage and surgical treatment were the main influencing factors for the survival time of female breast cancer patients.

Objective: To analyze the relationship between inflammatory factors and relevant risk factors in patients of essential hypertension (EH) combining acute coronary syndrome (ACS) with its clinical significance. Methods: Our research included 3 groups: EH group, n=79 patients with standard criteria, EH+ACS group, n=85 and Control group, n=48 normal subjects. Blood levels of lipoprotein-associated phospholipase A2 (Lp-PLA2), tryptase (TPS) and relevant clinical, biochemical parameters were measured; risk factors for cardiovascular disease were examined and the relationship between above parameters, risk factors and ACS occurrence in EH patients was studied by Logistic regression analysis. Results: The OR values were all greater than 1 in fibrinogen (Fbg), high-sensitivity C-reactive protein (hs-CRP), TPS, atherosclerotic plaque, Lp-PLA2 and EH grading. Fbg was the most significant independent risk factor (OR=22.242, 95% CI 6.458-76.609, P<0.0001), the standardized partial regression coefficient b'as absolute value (b') was 1.079 which was the highestone in above 6 variables with the strongest impact for ACS occurrence in EH patients. Conclusion: Fbg, hs-CRP, TPS, atherosclerotic plaque and EH grading were the independent risk factors for ACS occurrence in EH patients; Fbg was the highest risk factor for ACS occurrence with the strongest impact, which provided a new direction for ACS prevention and treatment.

BACKGROUNDTo construct and express a phage display library of anti human lung cancer monoclonal antibody 5F-11.

METHODSImmunoglobulin variable regions (VH,VL) were amplified from 5F-11 hybridrom by RT-PCR. ScFv genes consisting of VH DNA and VL DNA joined together by a linker DNA were cloned into a phage vector pCANTAB5E. After 4 rounds of screening with lung adenocarcinoma cell line A2 as antigen, an enriched secondary phage display library was obtained.

RESULTSA recombinant phage display library with total of 8×10⁷ pfu/ml was established. Randomized clones from unselected library digested with BstNⅠ showed different patterns, however, those from selected library showed that phages with special pattern were enriched. Twenty-three out of 30 clones were found to respond strongly to A2 cell lines.

CONCLUSIONSThe ScFv of anti-lung adenocarcinoma monoclonal antibody 5F-11 can be successfully produced, which may be useful to widen the application of the antibody.

BACKGROUNDTo study the effects of extraneous p53 antisense RNA on malignant growth and sensitivity to cisplatin of human lung cancer cell line.

METHODS801D cell line with p53 deletion and mutation at 248 codon was selected as a parent cell line. An 1.8 kb human p53 full length cDNA was inserted into a mammalian expression vector PEGFP to construct a p53 antisense RNA recombined plasmid PEGFP-p53(AS) and GFP gene at plasmid was a report gene to monitor extraneous gene expression. The extraneous gene was detected by PCR. The p53 mutation protein was examined by immunohitochemical stain of p53 monoclonal antibody. The inhibition growth efficacy of extraneous p53 in vitro was determined by clonogenic survival assay. Sensitivity of cells to cisplatin was examined with MTT assay. FCM analysis was performed to measure the effect of p53 antisense RNA on cell cycle.

RESULTSTwo cell lines, PEGFP-p53(AS)-801D and PEGFP-801D, were established after transfection of 801-D cells by lipofection and selection. Presence of extraneous p53 gene in PEGFP-p53(AS)-801D was proved by PCR and expression of extraneous p53 was estimated when green fluorescence in those cells was found out under the fluorescent microscopy. Mutated p53 protein in parent cell line 801D was positive and in PEGFP-p53(AS)-801D was negative with immunochemical stain. The inhibition rate of colony formation was 61% for PEGFP-p53(AS)-801D (P < 0.001). The sensitivity of PEGFP-p53(AS)-801D cells to cisplatin was increased. FCM analysis showed that the cell line was arrested at G1 phase.

CONCLUSIONSp53 mutation at 248 code plays an important role on malignant growth and resistance to cisplatin of human lung cancer cell line 801D. Malignant growth of cells with p53 deletion and mutation at 248 codon can be inhibited by extraneous p53 antisense RNA, and simultaneously the sensitivity to cisplatin is also increased.

BACKGROUNDTo isolate and clone the cisplatin genes in 801-D cell line, a kind of lung cancer cell line, with the emphasis of the objective genes regulated by wild type p53 (wtp53).

METHODSTotal RNA was extracted from transfected 801-D-wtp53, 801-D-vector cells which were both treated by cisplatin and 801-D-wtp53 cells. Using mRNA differential display, the DNA bands on gel were displayed by silver stain method. The DNA bands obtained from differential display were recovered and reamplified by PCR. The isolated genes were further proved by reverse Northern dot blot and were cloned to pGEMT easy vector.

RESULTSSix positive genes were identified and cloned. Out of them, 2 related fragments were found to have an open reading frame. One was partly homologous to ribonucleoside-diphosphate reductase A, and the other was no homologous to the known genes.

CONCLUSIONSThere are obvious differences in gene expression in 801-D-wtp53 after induced by cisplatin than two other controls. It is possible for p53 to regulate the sensitization of lung cancer cells to cisplatin through its downstream target genes.

OBJECTIVETo study synthesis of baicalin-copper and baicalin-aluminium complex and its antimicrobial, anti-tumor activity and anti-tumor effect against macrophages.

METHODBaicalin was reacted with metallic salt under a weak base condition to produce baicalin-copper and baicalin-aluminium complex. Baicalin and its synthesized complex were detected for antimicrobial activity against Staphylococcus aureus, Hay bacillus, Escherichia coli, Salmonella and Candida albicans by twofold broth dilution technique. Their anti-tumor activity against A549 and IC50 of HepG2 cells and anti-tumor effect against macrophages were detected by the MTT. And their phagocytic effect on macrophages was determined by the neutral red assay.

RESULTThe yields of baicalin-copper and baicalin-aluminium complex were 73.93% and 91.08%, respectively. The minimum inhibitory concentration (MIC) value against Staphylococcus aureus, Hay bacillus, Escherichia coli, Salmonella and Candida albicans was 0.0004, 0.0009, 0.0004, 0.0009, 0.000 4 mol x L(-1) for baicalin-copper complex and 0.0011, 0.0011, 0.0011, 0.0011, 0.0005 mol x L(-1) for baicalin-aluminium complex. The IC50 values against A549 and HepG2 cells were 89.6, 22.6 micromol x L(-1) for baicalin-copper complex, and 138.8, 97.2 micromol x L(-1) for baicalin-aluminium complex. The inhibitory ratio of macrophage on A549 cell was 43.52%, 80.89%, 52.66%, respectively, after the macrophages were stimulated by baicalin, baicalin-copper and baicalin-aluminium complex at a concentration of 160 micromol x L(-1).

CONCLUSIONThe acute toxicity test in mice showed that the complex was nontoxic to mice. Baicalin-copper complex showed the highest antimicrobial, anti-tumor activity, and the strongest effect on the anti-tumor activity of macrophage, while baicalin showed the lowest activities compared with baicalin-copper and baicalin-aluminium complex.

Aluminum ; Animals ; Anti-Bacterial Agents ; chemical synthesis ; chemistry ; pharmacology ; Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Copper ; Drugs, Chinese Herbal ; chemical synthesis ; chemistry ; pharmacology ; Flavonoids ; Humans ; Mice ; Microbial Sensitivity Tests

BACKGROUNDTo study the inhibition effects of both extraneous right sense and antisense p53 on malignant phenotype of human lung cancer cell line.

METHODSThe named 801D cell line with p53 deletion and mutation at 248 code was selected as a model in vitro. The recombined plasmid pEGFP-p53(RS) and pEGFP-p53(AS) were constructed. The extraneous gene was detected by PCR. The p53 mutation protein was examined by immunohistochemical stain of p53 antibody. The inhibition effect of extraneous p53 on tumor growth in vitro were determined by clonogenic survival assay. FCM analysis was carried out in cells. The inhibition effect on malignant growth of extraneous p53 in vivo was observed by heteroplastic transplant on nude mouse.

RESULTSThe transfected cell lines, pEGFP-p53(AS)-801D, pEGFP-p53(RS)-801D and pEGFP-801D were established. Presence of extraneous p53 and neo genes in pEGFP-p53(AS)-801D and pEGFP-p53(RS)-801D was proved by PCR and green fluorescence was found out in those cells under the microscope. Mutant protein in pEGFP-p53(AS)-801D was negative by immunohistochemical stain. The malignant growth of these transfected cell lines was inhibited comparing with parents in vivo and in vitro. Inhibition rate of colony formation was 62.0% for pEGFP-p53(AS)-801D and 80.8% for pEGFP-p53(RS)-801D. The tumorigenicity in nude mice was suppressed. Inhibition effects of extraneous right sense p53 on malignant growth of 801D was more distinct. FCM analysis showed that pEGFP-p53(AS)-801D cells were arrested at G1 phase.

CONCLUSIONSThe transfected cell lines with extraneous right sense and antisense p53 appear that malignant growth can be inhibited in vivo and in vitro.

BACKGROUND AND OBJECTIVEThe p53 as a transcription factor in cell stress was activated to regulate cell cycle and programmed cell death to inhibit tumor growth. Usually, p53 is kept in non-activated state through various mechanisms, including the action of p53 C-terminal negative regulatory sequences. The purpose of the study is to prepare the two types p53 recombinant adenoviruses that carry full-length p53 as well as deletion of negative regulatory sequences at p53 C-terminus and to detect exogenous GFP expression in human lung cancer cell infected-virus by FCM scatter plot.

METHODSUsing pAdEasy-Track vector system the p53 recombinant plasmids was constructed and the homologous recombinants in E. coli was produced. The three kinds of recombinant adenovirus in L293 cells was generated, sequencing proved. Exogenous GFP expression in human lung cancer 801D cells infected-virus was detected by FCM scatter plot.

RESULTSp53 recombinant adenoviruses named Ad-p53(wtp), Ad-p53(del) and Ad-(empty carrier) were produced. Results of sequences indicate that the Ad-p53(del) was deletion of 111 bases before stop codon TGA and of 3 untranslated region at p53, the Ad-p53(wtp) no loss of any p53 base, the Ad-(empty carrier) no p53 sequence. FCM scatter plot indicate the percentage of 801D cells expressed GFP with three kinds of viral infection was almost same and was increased with the virus density. 801D contains ratio of cells with different fluorescence intensity.

CONCLUSIONThe preparation of recombinant adenovirus, Ad-p53(del), pA-p53(wtp) and Ad-(empty carrier). The cells expressed-GFP can be quantitatively detected by FCM scatter plot. It was provide that the reliability of the virus system and accurate method for selecting viruses density to infecting cells.

Adenoviridae ; genetics ; Animals ; Flow Cytometry ; methods ; Genes, p53 ; Green Fluorescent Proteins ; genetics ; Humans ; Mice ; Recombination, Genetic

Objective:To investigate the molecular mechanisms of chronic rhinosinusitis (CRS), to identify key cell subgroups and genes, to construct effective diagnostic models, and to screen for potential therapeutic drugs.Methods:Key cell subgroups in CRS were identified through single-cell transcriptomic sequencing data. Essential genes associated with CRS were selected and diagnostic models were constructed by hdWGCNA (high dimensional weighted gene co-expression network analysis) and various machine learning algorithms. Causal inference analysis was performed using Mendelian randomization and colocalization analysis. Potential therapeutic drugs were identified using molecular docking technology, and the results of bioinformatics analysis were validated by immunofluorescence staining. Graphpad Prism, R, Python, and Adobe Illustrator software were used for data and image processing.Results:An increased proportion of basal and suprabasal cells was observed in CRS, especially in eosinophilic CRS with nasal polyps (ECRSwNP), with P=0.001. hdWGCNA revealed that the "yellow module" was closely related to basal and suprabasal cells in CRS. Univariate logistic regression and LASSO algorithm selected 13 key genes ( CTSC, LAMB3, CYP2S1, TRPV4, ARHGAP21, PTHLH, CDH26, MRPS6, TENM4, FAM110C, NCKAP5, SAMD3, and PTCHD4). Based on these 13 genes, an effective CRS diagnostic model was developed using various machine learning algorithms (AUC=0.958). Mendelian randomization analysis indicated a causal relationship between CTSC and CRS (inverse variance weighted: OR=1.06, P=0.006), and colocalization analysis confirmed shared genetic variants between CTSC and CRS (PPH4/PPH3>2). Molecular docking results showed that acetaminophen binded well with CTSC (binding energy:-5.638 kcal/mol). Immunofluorescence staining experiments indicated an increase in CTSC +cells in CRS. Conclusion:This study integrates various bioinformatics methods to identify key cell types and genes in CRS, constructs an effective diagnostic model, underscores the critical role of the CTSC gene in CRS pathogenesis, and provides new targets for the treatment of CRS.