Objective The effectiveness of Zhuang nationality medical lotus needle plus back cupping therapy ( Zhuang needle-cupping therapy) , Flixonase aqueous nasal spray and cetirizine tablets in treating allergic rhinitis (AR) was compared for the exploration of the therapeutic mechanism of Zhuang needle-cupping therapy. Methods A total of 200 recruited AR patients were randomly divided into four groups in the proportion of 1:1:1:1. The four groups were Zhuang needle-cupping therapy group, cetirizine group, Flixonase group and blank control group. The blank control group had no medication, and the patients of the other three medication groups were given the corresponding treatment. Ten days constituted one treatment course, and interval between two courses lasted one week. After two courses, the therapeutic effect was evaluated. The changes of specific IgE (S-IgE), leukotriene (LT), interleukin 4(IL-4), IL-9 mRNA, interferon gamma (IFN-γ), Thl / Th2 cells, and Th17 cytokine ( IL-17) were observed before and after treatment. Results ( 1) After two treatment courses, Zhuang needle-cupping therapy group had better therapeutic effect than cetirizine group , Flixonase group and blank control group, and the therapeutic effect of cetirizine group and Flixonase group was better than the blank control group (P<0.05). However, the therapeutic effect of cetirizine group was similar to that of Flixonase group ( P>0.05). ( 2) After treatment, the levels of S-IgE, LT, IL-9 mRNA, IL-4 and IL-17 were decreased, and IFN-γ and Th1/Th2 levels were increased in the three medication groups ( P<0.05 compared with those before treatment) . The differ ences of the laboratory indexes in the blank control group were insignificant before and after treatment ( P>0.05). The results of inter-group comparison after treatment showed that Zhuang needle-cupping therapy group had better effect on improving S-IgE, LT, IFN-γand Th1/Th2 than cetirizine group and Flixonase group (P<0.05). (3) During the trial, no adverse reaction was found. Conclusion Zhuang needle-cupping therapy exerts certain therapeutic effect for AR, and the mechanism may be related with the inhibition of S-IgE, LT, IL-9 mRNA and IL-17 expression, and with the regulation of Th1/Th2 imbalance by decreasing TH2 cytokine level and increasing Th1 cytokine level.

This research used inter simple sequence repeat(ISSR)markers to analyze the genetic diversity of Hedyotis diffusa Willd. from different origins. A total of 23 samples of Hedyotis diffusa Willd. in the Guangxi Zhuang Autonomous Region, Guangdong, Hunan, Zhejiang, Fujian, Jiangxi and Anhui, respectively were collected. 150 ISSR primers were used to amplify PCR and then POPGENE1. 32, NTSYS2. 10 software were used to analyze genetic diversity. 11 primers were screened, 115 polymorphic bands were amplified, the polymorphism ratio was 85. 22%, the number of alleles(Na)was 1. 852 2, the effective allele(Ne)was 1. 543 4, Neis gene diversity index(H)was 0. 316 5 and Shannon′s information index(I)was 0. 470 0. The results of cluster analysis show that the Hedyotis diffusa can be divided into three clades. The conclusion is that ISSR molecular markers can provide a insight for the identification of Hedyotis diffusa Willd. .

OBJECTIVETo study the gene mutation and streptomycin, isoniazid or rifampicin resistance of Mycobacterium isolated from silico-tuberculosis patient's sputum so as to find a more effective therapy for this disease.

METHODSMycobacteria tuberculosis were separated from 96 coal worker with silico-tuberculosis firstly. Then rpsL, KatG and rpoB fragments of genome were copied with PCR and compared their SSCP profiles with standard strains.

RESULTS67 strains of streptomycin, isoniazid or rifampicin resistant Mycobacteria tuberculosis were found in routine drug resistance test, with the percentages of 80.5% (54/67), 58.2% (39/67) respectively. PCR-SSCP showed that out of 67 drug-resistant strains, 66(98.5%) of rpsL, 47(70.1%) of rpoB and 42(62.7%) of KatG appeared abnormal.

CONCLUSIONMost of the resistant strains appeared gene mutation. The mution rates were higher than the results from routine drug resistance test.

Coal ; Drug Resistance, Bacterial ; Humans ; Mutation ; Mycobacterium tuberculosis ; drug effects ; genetics ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Silicotuberculosis ; microbiology ; Sputum ; microbiology

Background: African swine fever virus (ASFV), classical swine fever virus (CSFV), and porcine reproductive and respiratory syndrome virus (PRRSV) are still prevalent in many regions of China. Co-infections make it difficult to distinguish their clinical symptoms and pathological changes. Therefore, a rapid and specific method is needed for the differential detection of these pathogens. Objectives: The aim of this study was to develop a multiplex real-time quantitative reverse transcription polymerase chain reaction (multiplex qRT-PCR) for the simultaneous differential detection of ASFV, CSFV, and PRRSV. Methods: Three pairs of primers and TaqMan probes targeting the ASFV p72 gene, CSFV 5′untranslated region, and PRRSV ORF7 gene were designed. After optimizing the reaction conditions, including the annealing temperature, primer concentration, and probe concentration, multiplex qRT-PCR for simultaneous and differential detection of ASFV, CSFV, and PRRSV was developed. Subsequently, 1,143 clinical samples were detected to verify the practicality of the assay. Results: The multiplex qRT-PCR assay could specifically and simultaneously detect the ASFV, CSFV, and PRRSV with a detection limit of 1.78 × 10 0 copies for the ASFV, CSFV, and PRRSV, but could not amplify the other major porcine viruses, such as pseudorabies virus, porcine circovirus type 1 (PCV1), PCV2, PCV3, foot-and-mouth disease virus, porcine parvovirus, atypical porcine pestivirus, and Senecavirus A. The assay had good repeatability with coefficients of variation of intra- and inter-assay of less than 1.2%. Finally, the assay was used to detect 1,143 clinical samples to evaluate its practicality in the field. The positive rates of ASFV, CSFV, and PRRSV were 25.63%, 9.36%, and 17.50%, respectively. The co-infection rates of ASFV+CSFV, ASFV+PRRSV, CSFV+PRRSV, and ASFV+CSFV+PRRSV were 2.45%, 2.36%, 1.57%, and 0.17%, respectively. Conclusions The multiplex qRT-PCR developed in this study could provide a rapid, sensitive, specific diagnostic tool for the simultaneous and differential detection of ASFV, CSFV, and PRRSV.

Objective:To investigate the effects of different doses of oxiracetam (ORC) on the neurological impairment and oxidative stress ability of lead(Pb)-exposed rats.Methods:Total 32 male SD rats with SPF grade were randomly divided into control group, the lead-exposed group, low-dose ORC intervention group and high-dose ORC intervention group according to the random number table method, with 8 rats in each group.The neurobehavioral indexes of rats were measured by gait score, tail flicking test, hindlimb support test and Morris water maze test.The lead content in hippocampal tissue was detected by spectrophotometry.The cell morphology of hippocampal tissue was observed by HE staining.Superoxide dismutase (superoxide dismutase, SOD) level in hippocampal tissues was detected by xanthine oxidase, and the level of malondialdehyde (MDA) in the hippocampus tissue was detected by the thiobarbiturate, and the level of glutathione peroxidase (GPx) in the hippocampus tissue was detected by chemical colorimetric.SPSS 24.0 software was used for statistical analysis, one-way ANOVA was used for multiple group comparisons and LSD- t test was used for further pairwise comparisons. Results:(1)After 8 weeks of lead exposure, there was no significant difference in body weight among the 4 groups( F=0.869, P=0.469). (2)Results of neurobehavioral indicators: there were statistically significant differences in gait scores, tail flick time, hind limb deployment distance, escape latency, and number of crossing platforms among the four groups of rats ( F=7.854, 13.630, 8.484, 23.485, 45.457, all P<0.05). The gait score, tail flick time, hind limb deployment distance, and escape latency of the lead-exposed group rats were higher than those of the control group (all P<0.05), while the number of crossing platforms was lower than that of the control group ( P<0.05). The gait score, tail flick time, hind limb deployment distance, and escape latency of the high-dose ORC intervention group were lower than those of the lead exposed group (all P<0.05), and the number of crossing platforms was higher than that of the lead exposed group ( P<0.05). (3)Lead content in hippocampal tissue: there was a statistically significant difference in lead content in the hippocampus of the four groups( F=309.013, P<0.001). The lead contents of lead exposed group ((1.21±0.10)μg/g), low-dose ORC intervention group ((1.03±0.10)μg/g) and high-dose ORC intervention group ((1.02±0.06)μg/g) were higher than that of the control group((0.02±0.00) μg/g) (all P<0.05), while the lead content in the low-dose ORC intervention group and high-dose ORC intervention group were both lower than that of the lead exposed group (both P<0.05). (4) HE staining showed that compared with the control group, the hippocampal tissue cells in the lead exposed group were arranged disordered, the tissue was loose, and the number of cells was reduced.Compared with the lead exposed group, the hippocampal histiocytes were closely arranged and regular, and the nuclei were fuller.(5)Oxidative stress levels in hippocampal tissue: there were significant differences in MDA, GPx content and SOD activity of hippocampal tissues in the four groups( F=69.879, 56.757, 11.644, all P<0.001). The levels of SOD ((2.03±0.18)U/mg, (3.42±0.26)U/mg), GPx((67.29±7.94)nmol/mg, (89.50±7.94)nmol/mg) in the hippocampus tissue of the lead exposed group were lower than those of the control group (all P<0.05), while the content of MDA was higher than that of the control group((43.73±3.74) nmol/mg, (16.42±1.60) nmol/mg)( P<0.05). The levels of SOD ((3.32±0.12) U/mg) and GPx ((84.11±6.26) nmol/mg) in the high-dose ORC intervention group were higher than those in the lead exposed group (both P<0.05), while the levels of MDA ((21.05±2.56) nmol/mg) was lower than that in the lead exposed group ( P<0.05). Conclusion:ORC can alleviate neurological damage in rats caused by lead exposure, which may be related to the up-regulation of antioxidant capacity of hippocampal tissues, thereby improving pathological damage.

Objective:To investigate the iodine nutritional status of pregnant women in Jilin Province and its correlation with the distribution characteristics of water iodine in external environment, providing a basis for scientific iodine supplementation and prevention of iodine deficiency disorders.Methods:A retrospective analysis was conducted on the iodine survey data of drinking water for residents in Jilin Province in 2017 and the monitoring data of iodine deficiency disorders in 2021. The water iodine, salt iodine, and urinary iodine level of pregnant women were analyzed.Results:In 8 866 water samples from 873 townships (streets, hereinafter referred to as townships) of 60 counties (cities, districts) in 9 cities (autonomous prefectures) throughout the province, the median of water iodine was 4.60 μg/L, ranging from 0.00 to 81.30 μg/L. Among them, there were 758 townships with a median water iodine < 10 μg/L, accounting for 86.83% (758/873); 107 townships with a water iodine of 10 - < 40 μg/L, accounting for 12.26% (107/873). The median salt iodine was 23.50 mg/kg in 6 000 household consumption salt samples. The iodized salt coverage rate, iodized salt qualified rate, and qualified iodized salt consumption rate were 99.50% (5 970/6 000), 97.30% (5 809/5 970), and 96.82% (5 809/6 000), respectively. The iodized salt coverage rate in 9 cities (autonomous prefectures) were > 95%, the iodized salt qualified rate and qualified iodized salt consumption rate were > 90%. The median urinary iodine in 6 000 pregnant women's urine samples was 169.05 μg/L. Except for Bayshan City, which was iodine-deficient, the other 8 cities (autonomous prefectures) were iodine-suitable. The results of correlation analysis showed that there was no correlation between the urinary iodine level of pregnant women and the distribution of water iodine in the external environmental at the municipal (autonomous prefecture) level ( r = 0.60, P = 0.089). Conclusions:Most townships in Jilin Province are iodine-deficient in the external environment, and there are no water-borne high iodine area. The iodized salt coverage rate, iodized salt qualified rate, and qualified iodized salt consumption rate all meet the national standards. The iodine nutrition of pregnant women is generally at a suitable level, but there are still some areas where pregnant women are iodine-deficient, and there is no correlation with the distribution of water iodine.

Objective:Establishment of a new model of human primary colon cancer transplantation tumor in normal immune mice and to provide a reliable experimental animal model for studying the pathogenesis of colon cancer under normal immunity.Methods:Human colon cancer cells come from colon cancer patients who underwent surgery in the Affiliated Hospital of Jining Medical College in 2017. The mice in the cell control group were inoculated with phosphate buffered solution (PBS) containing colon cancer cells, the microcarrier control group was inoculated with PBS containing microcarrier 6, and the cell-microcarrier complex group was inoculated with the PBS containing colon cancer cell-microcarrier complex. The cells of each group were inoculated under the skin of the right axilla of mice by subcutaneous injection, and the time, size, tumor formation rate and pathological changes under microscope were recorded. The transplanted tumor tissue was immunohistochemically stained with the EnVisiion two-step method, and the tumor formation rate of the transplanted tumor was judged according to the proportion of positive cells in the visual field. The polymerase chain reaction (PCR) method was used to detect the expression of human-specific Alu sequence in mice tumor tissue.Results:After inoculation with tumor cells, the mice in the cell control group and the microcarrier control group did not die and did not form tumors; the mice in the cell-microcarrier complex group had palpable subcutaneous tumors in the right axillary subcutaneously on the 5th to 7th days after inoculation, and tumor formation rate is 67% (10/15), and the tumor volume can reach about 500 mm 3 2 to 3 weeks after vaccination. The immunohistochemistry results showed that CK20, CDX-2 and carcinoembryonic antigen were all positively expressed. The PCR results showed that the expression of human-specific Alu sequence can be detected in the transplanted tumor tissue of tumor-bearing mice. Conclusion:Human primary colon cancer cells used microcarrier 6 as a carrier to form tumors in normal immunized mice, and successfully established a new model of human colon cancer transplantation tumor in normal immune mice.

Objective:Establishment of a new model of human primary colon cancer transplantation tumor in normal immune mice and to provide a reliable experimental animal model for studying the pathogenesis of colon cancer under normal immunity.Methods:Human colon cancer cells come from colon cancer patients who underwent surgery in the Affiliated Hospital of Jining Medical College in 2017. The mice in the cell control group were inoculated with phosphate buffered solution (PBS) containing colon cancer cells, the microcarrier control group was inoculated with PBS containing microcarrier 6, and the cell-microcarrier complex group was inoculated with the PBS containing colon cancer cell-microcarrier complex. The cells of each group were inoculated under the skin of the right axilla of mice by subcutaneous injection, and the time, size, tumor formation rate and pathological changes under microscope were recorded. The transplanted tumor tissue was immunohistochemically stained with the EnVisiion two-step method, and the tumor formation rate of the transplanted tumor was judged according to the proportion of positive cells in the visual field. The polymerase chain reaction (PCR) method was used to detect the expression of human-specific Alu sequence in mice tumor tissue.Results:After inoculation with tumor cells, the mice in the cell control group and the microcarrier control group did not die and did not form tumors; the mice in the cell-microcarrier complex group had palpable subcutaneous tumors in the right axillary subcutaneously on the 5th to 7th days after inoculation, and tumor formation rate is 67% (10/15), and the tumor volume can reach about 500 mm 3 2 to 3 weeks after vaccination. The immunohistochemistry results showed that CK20, CDX-2 and carcinoembryonic antigen were all positively expressed. The PCR results showed that the expression of human-specific Alu sequence can be detected in the transplanted tumor tissue of tumor-bearing mice. Conclusion:Human primary colon cancer cells used microcarrier 6 as a carrier to form tumors in normal immunized mice, and successfully established a new model of human colon cancer transplantation tumor in normal immune mice.

Objective A competing endogenous RNA (ceRNA) regulatory network associated with long non-coding RNA (lncRNA) specific for lung adenocarcinoma (LUAD) was constructed based on bioinformatics methods, and the functional mechanism of actinfilament-associated protein 1-antisense RNA1 (AFAP1-AS1) in LUAD was analyzed, in order to provide a new direction for the study of LUAD therapeutic targets. Methods The gene chip of LUAD was downloaded from the Gene Expression Omnibus (GEO), and lncRNA and mRNA with differential expression between LUAD and normal tissues were screened using GEO2R online software, and their target genes were predicted by online databases to construct ceRNA networks and perform enrichment analysis. In cell experiments, AFAP1-AS1 was genetically knocked down and siRNA was constructed and transfected into LUAD cells A549 by cell transfection. CCK8, transwell, scratch assay and flow cytometry were used to detect the ability of cells to proliferate, invade, migrate and apoptosis. Results A total of 6 differentially expressed lncRNA and 494 differentially expressed mRNA were identified in the microarray of LUAD. The ceRNA network involved a total of 6 lncRNA, 22 miRNA, and 55 mRNA. Enrichment analysis revealed that mRNA was associated with cancer-related pathways. In cell assays, knockdown of AFAP1-AS1 inhibited cell proliferation, invasion, and migration, and AFAP1-AS1 promoted apoptosis. Conclusion In this study, we construct a lncRNA-mediated ceRNA network, which may help to further investigate the mechanism of action of LUAD. In addition, through cellular experiments, AFAP1-AS1 is found to have potential as a therapeutic target for LUAD.

BACKGROUNDCytochrome b5 reductase 2 (CYB5R2) is a potential tumor suppressor that inhibits cell proliferation and motility in nasopharyngeal carcinoma (NPC). Inactivation of CYB5R2 is associated with lymph node metastasis in NPC. This study aimed to explore the mechanisms contributing to the anti-neoplastic effects of CYB5R2.

METHODSPolymerase chain reaction (PCR) assays were used to analyze the transcription of 84 genes known to be involved in representative cancer pathways in the NPC cell line HONE1. NPC cell lines CNE2 and HONE1 were transiently transfected with CYB5R2, and data was validated by real-time PCR. A chick chorioallantoic membrane (CAM) embryo model was implanted with CYB5R2-expressing CNE2 and HONE1 cells to evaluate the effect of CYB5R2 on angiogenesis. An immunohistochemical assay of the CAM model was used to analyze the protein expression of vascular endothelial growth factor (VEGF).

RESULTSIn CYB5R2-transfected NPC cells, PCR assays revealed up-regulated mRNA levels of Fas cell surface death receptor (FAS), FBJ murine osteosarcoma viral oncogene homolog (FOS), phosphoinositide-3-kinase regulatory subunit 1 (PIK3R1), integrin beta 3 (ITGB3), metastasis suppressor 1 (MTSS1), interferon beta 1 (IFNB1), and cyclin-dependent kinase inhibitor 2A (CDKN2A) and down-regulated levels of integrin beta 5 (ITGB5), insulin-like growth factor 1 (IGF1), TEK tyrosine kinase (TEK), transforming growth factor beta receptor 1 (TGFBR1), and VEGF. The angiogenesis in the CAM model implanted with CYB5R2-transfected NPC cells was inhibited. Down-regulation of VEGF by CYB5R2 in NPC cells was confirmed by immunohistochemical staining in the CAM model.

CONCLUSIONCYB5R2 up-regulates the expression of genes that negatively modulate angiogenesis in NPC cells and down-regulates the expression of VEGF to reduce angiogenesis, thereby suppressing tumor formation.

Animals ; Carcinoma ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Chickens ; Cytochrome-B(5) Reductase ; Down-Regulation ; Gene Regulatory Networks ; Genes, Tumor Suppressor ; Humans ; Nasopharyngeal Neoplasms ; Neovascularization, Pathologic ; Oxidoreductases ; Real-Time Polymerase Chain Reaction ; Transfection ; Up-Regulation ; Vascular Endothelial Growth Factor A