AIM : To explore the effects of ASODN targeting survivin on apoptosis in HL 60 cells. METHODS : The cell proliferation was tested by MTT assay, the cell morphological transformation was observed by inverted microscope, the cell apoptosis index was examined by TUNEL, the cell apoptosis rate was precisely detected by flow cytometry, and the expression of survivin mRNA was detected by reverse transcriptase PCR. RESULTS : 5-20 ?mol?L -1 of survivin ASODN showed obviously inhibitory effect on the cell proliferation of HL 60 cells, and the inhibitory effect correlated with time and dosage. The cell apoptosis rate and the expression inhibitory rate of survivin gene in ASODN groups were obviously higher than that in the control group. Effects of antisense oligonucleotide targeting survivin on apoptosis in HL 60 cells$$$$ QI Shi mei, BI Fu yong Department of Biochemistry, Wannan Medical College, Wuhu 241001, Anhui, China ABSTRACT AIM : To explore the effects of ASODN targeting survivin on apoptosis in HL 60 cells. METHODS : The cell proliferation was tested by MTT assay, the cell morphological transformation was observed by inverted microscope, the cell apoptosis index was examined by TUNEL, the cell apoptosis rate was precisely detected by flow cytometry, and the expression of survivin mRNA was detected by reverse transcriptase PCR. RESULTS : 5-20 ?mol?L -1 of survivin ASODN showed obviously inhibitory effect on the cell proliferation of HL 60 cells, and the inhibitory effect correlated with time and dosage. The cell apoptosis rate and the expression inhibitory rate of survivin gene in ASODN groups were obviously higher than that in the control group. [WTHZ CONCLUSION : ASODN targeting survivin can effectively inhibit the expression of survivin mRNA and induce the cell apoptosis, and it indicates that survivin plays an important role in maintaining the proliferation of tumor cells.

Objective To explore β-sodium aescinate on vascular endothelial function ( FMD ) , homocysteine ( Hcy ) and hypersensitive C-reactive protein ( hs-CRP) and clinical efficacy in patients with acute cerebral infarction.Methods 198 acute cerebral infarction patients from March 2013 to April 2015 were randomly divided into observation group (n=100) and control group (n=98).Control group were treated according to the condition of the disease, observation group were treated by β-sodium aescinate base on control group, 20mg was added to 250mL saline for intravenous drip,one times per day.Continuous used 14d for one treatment courses.Compared the change of vascular endothelial function, Hcy and hs-CRP and clinical efficacy.Results The total effective rate of observation group was 90.00%, which was significantly higher than that of 71.42% in control group (χ2 =11.01,P<0.05).Post-treatment the value of FMD significantly increased, Hcy and hs CRP were significantly decreased both in observation group and control group respectively, which the difference had a statistically significant as compared with Pre-treatment (P<0.05);but, the value of FMD was significantly higher, Hcy and hs CRP was significantly lower in observation group than that of control group (P<0.05).Conclusion It has a significant β-sodium aescinate clinical effect in treatment of acute cerebral infarction, and FMD are significantly higher, Hcy and hs-CRP are significantly decrease.

OBJECTIVETo investigate the dual role of daphnetin in suppressing high mobility group box-1 protein (HMGB1) release and blocking HMGB1-induced inflammatory response.

METHODSMurine macrophage RAW264.7 cells were cultured in the presence of daphnetin, lipopolysaccharide (LPS), or both. HMGB1 release from the cells was determined using ELISA, and phosphorylations of JAK1/2 and of STAT1 were detected by Western blotting. Human monocytic THP-1 cells exposed to daphnetin, rhHMGB1, or both were examined for NO production using a NO detection kit, for the release of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and prostaglandin E2 (PGE2) using ELISA, and for expressions of iNOS, COX-2 and phosphorylated p38, ERK, and JNK with Western blotting.

RESULTSDaphnetin dose-dependently reduced the release of HMGB1 in RAW264.7 cells and suppressed rhHMGB1-induced iNOS and COX-2 expressions and release of TNF-α, IL-6, PGE2, and NO in THP-1 cells. Western blotting revealed that daphnetin significantly down-regulated the phosphorylations of JAK-STAT1 pathway in LPS-stimulated RAW264.7 cells but did not suppress the phosphorylations of MAPKs signaling pathway induced by rhHMGB1 in THP-1 cells.

CONCLUSIONDaphnetin can reduce the release of HMGB1 and suppress HMGB1-induced inflammatory response. In RAW264.7 cells, daphnetin inhibited LPS induced HMGB1 release is at least partly mediated by suppressing JAK-STAT1 signaling pathway activation.

Animals ; Cell Line ; Cyclooxygenase 2 ; metabolism ; Dinoprostone ; metabolism ; HMGB1 Protein ; metabolism ; Humans ; Inflammation ; metabolism ; Interleukin-6 ; metabolism ; Janus Kinase 1 ; metabolism ; Lipopolysaccharides ; Macrophages ; drug effects ; Mice ; Monocytes ; drug effects ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; RAW 264.7 Cells ; STAT1 Transcription Factor ; metabolism ; Signal Transduction ; drug effects ; Tumor Necrosis Factor-alpha ; metabolism ; Umbelliferones ; pharmacology

OBJECTIVE: To investigate the inhibitory effects of dihydromyricetin on the proliferation and migration of gastric cancer BGC-823 cells and explore the molecular mechanisms. METHODS: BGC-823 cells in routine culture were treated with different concentrations of dihydromyricetin (0, 40, 60, 80, 100, and 120 μg/mL) for 24 h, and the changes in cell viability were detected using CCK-8 assay; colony forming assay and Transwell assay were performed to assess the changes in colonyforming and migration abilities of the cells, respectively. The levels of MMP-2 and MMP-9 in the treated cells were determined using ELISA, and Western blotting was used to detect the expressions of E-cadherin, N-cadherin, cyclin D1, cyclin E1, HSP70 and HMGB1 and the phosphorylation levels of Akt and Stat3. RESULTS: CCK-8 assay showed that dihydromyricetin treatment dose-dependently inhibited the viability of BGC-823 cells ( CONCLUSIONS Dihydromyricetin inhibits the proliferation and migration of BGC-823 cells through suppressing the activation of Akt/stat3 signaling pathways and HMGB1 expression.
Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Flavonols ; HMGB1 Protein/metabolism* ; Humans ; Proto-Oncogene Proteins c-akt/metabolism* ; STAT3 Transcription Factor ; Stomach Neoplasms

ObjectiveTo analyze the epidemic characteristics of dengue fever in Mengla County and provide basis for scientific prevention and control of dengue fever. MethodsWe collected the case information of dengue fever in Mengla County reported by the infectious disease reporting information system of China Center for Disease Control and prevention from January 1 to December 31， 2019 and the case field investigation records. The case data were analyzed by descriptive epidemiological method. ResultsIn 2019， Mengla County reported 369 cases of dengue fever， all of which were unclassified， including 354 clinically diagnosed cases， 15 confirmed cases， 6 severe cases， and there was no deaths. The annual incidence rate was 120.98/10⁵. Mengla Town had the most cases （145 cases， 39.30%） followed by 63 cases （17.07%） in Mengpeng Town. The reported cases were mainly local cases （65.85%）. The ratio of male to female was 1.25∶1. The age distribution was mainly in the group of 21‒60 years old （82.38%）. Farmers （112 cases， 30.35%） and business service providers （85 cases， 23.04%） were the majority. The annual cases were distributed from May to November， of which the most were reported in September， and the number of cases reported from July to October accounts for 93.22% of all cases. ConclusionMengla County is still a high incidence area of dengue fever in Yunnan Province， and the vector Aedes is widespread. It is suggested to strengthen mosquito prevention and control in the epidemic season， actively carry out patriotic health campaign， carry out special rectification of the environment in rural areas， and conduct effective public education.

OBJECTIVE: To investigate the effect of calenduloside E on lipopolysaccharide (LPS)-induced inflammatory response in RAW264.7 cells and explore the underlying molecular mechanism. METHODS: CCK-8 assay was used to examine the effect of different concentrations of calenduloside E (0-30 μg/mL) on the viability of RAW264.7 cells. The release of the pro-inflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in RAW264.7 cells in response to pretreatment with 6, 8, and 10 μg/mL calenduloside E for 2 h followed by stimulation with 100 ng/mL LPS was detected using enzyme-linked immunosorbent assay (ELISA). The expression levels of iNOS and COX-2 and the activation of JAK-stats, MAPKs and NF-кB signaling pathways in the treated cells were determined using Western blotting. A reactive oxygen species (ROS) detection kit was used to detect ROS production in the cells, and the nuclear translocation of the transcription factor stat3 was observed by laser confocal microscopy. RESULTS: Calenduloside E below 20 μg/mL did not significantly affect the viability of RAW264.7 cells. Calenduloside E dose-dependently decreased the expression levels of iNOS and COX-2 induced by LPS, inhibited LPS-induced release of TNF-α and IL-1β, and suppressed LPS-induced JAK1-stat3 signaling pathway activation and stat3 nuclear translocation. Calenduloside E also significantly reduced ROS production induced by LPS in RAW264.7 cells. CONCLUSIONS Calenduloside E inhibits LPS-induced inflammatory response by blocking ROS-mediated activation of JAK1-stat3 signaling pathway in RAW264.7 cells.
Animals ; Lipopolysaccharides ; Mice ; NF-kappa B ; Oleanolic Acid ; analogs & derivatives ; RAW 264.7 Cells ; Reactive Oxygen Species ; Saponins ; Signal Transduction

ObjectiveTo analyze the epidemic characteristics and trend of mosquito vectors related to Japanese encephalitis （JE） in Mengla County， and to provide scientific evidence for JE prevention and control. MethodsThe JE related mosquito vector monitoring data in Mengla County from 2016 to 2020 were collected and subjected to further statistical analysis. ResultsA total of 1 689 mosquitoes were captured at the JE mosquito vector monitoring sites in Mengla County， 36.3% of which were captured in 2020 and 13.3% in 2017. The density of Culex tritaeniorhynchus was the highest （3.04 per lamp per day）， and that of Anopheles sinensis was the lowest （1.03 per lamp per day）. The distribution of mosquito species showed significant difference in the same year. The mosquito density in pig house was 12.93 per lamp∙day， and that in human house was 4.67 per lamp∙day. The mosquito density of different mosquito species in pig house was higher than that in human house. There was no significant difference in the site distribution of Anopheles sinensis， but there were significant differences in the site distribution of Culex tritaeniorhynchus， Culex pipiens quinquefasciatus and other mosquito species. The mosquito density peaked in May （12.78 per lamp per day） and July （10.28 per lamp per day）. The temporal distribution showed that the vector density decreased gradually from May to October， however， the species population structure also varied significantly， and the trends of each mosquito species also varied greatly. In Mengla， Culex tritaeniorhynchus peaked in May. ConclusionThe JE epidemic situation in Mengla County is still severe. It is recommended to strengthen prevention and control in the peak season of mosquito activities， such as actively carrying out patriotic health campaigns， and effective public education. At the same time， we should also strengthen the JE vaccination for school-age children and the training of medical personnel.

OBJECTIVETo study the effect of chrysin in inducing apoptosis of human hepatic carcinoma cells and explore the possible mechanism.

METHODSHuman hepatic carcinoma SMMC-7721 cells treated with DMSO or chrysin at different concentrations (5-200 μg/mL) were examined for changes in the cell proliferation using CCK-8 assay. The morphological changes of SMMC-7721 cells were observed in response to treatment with 5, 10, or 20 μg/mL chrysin, and the changes in the cell nuclei were observed using DAPI nuclear staining. Annexin Ⅴ-FITC/PI flow cytometry was used to determine the cell apoptosis rate. The changes in the apoptosis-related proteins (PARP and caspase-3) and MAPKs signal pathway were detected with Western blotting.

RESULTSChrysin treatment obviously suppressed the proliferation of SMMC-7721 cells in a dose-dependent manner below the concentration of 60 μg/mL. Chrysin (20 μg/mL) also caused significantly increased cell apoptosis and significant cleavage of PARP and caspase-3. Chrysin significantly activated MAPKs signaling pathway in a time-and dose-dependent manner, with the peak activation level occurring at 15 min. Pretreatment of the cells with specific inhibitors of the MAPKs pathway obviously inhibited the effect of chrysin in inducing cell apoptosis.

CONCLUSIONSChrysin inhibits the proliferation and promotes apoptosis of SMMC-7721 cells by regulating the activation of MAPKs signaling.

OBJECTIVETo understand the one-year effect of HCV/HIV co-infected patients who had received AIDS second-line antiretroviral treatment after failure virologically, on the first-line therapy.

METHODSHCV and HIV antibody positive patients who had experienced virological failure but received at least one-year AIDS first-line treatment, were recruited from May to October 2012 in Xincai, Queshan and Weishi of Henan province. 6-months and 12-months follow-up programs were carried out after the regimen had been changed to AIDS second-line antiretroviral treatment, CD4⁺ T lymphocyte count, HIV-1 virus load and HIV-1 drug resistance were performed.

RESULTSEighty-one cases of eligible patients were selected and followed by an amelioration of CD4 median at 6-month and 12-month follow-up period. Data showed that the baseline, 6-months and 12-months CD4 medians were 266 cells/µl, 275 cells/µl and 299 cells/µl (χ² = 8.214, P = 0.009). The ratio of HIV virus load suppression patients at 6-months and 12-months follow-up increased to 46.84% and 50.00%, respectively. Frequencies of HIV drug resistance also decreased at the baseline, 6-months and 12-months, with ratios as 66.67%, 26.58% and 27.63% (χ² = 29.362, P = 0.000), respectively. Ratios of patients that holding NRTI and NNRTI drug resistance appeared coinstantaneous decrease at the baseline, 6-months and 12-months, as 51.85%, 18.99% and 17.11% (χ² = 14.230, P = 0.005). At the baseline, the ratios of patients resisted to 3TC, ABC and FTC were all more than 50%, with AZT, D4T and DDI between 41%-44% while TDF appeared as 33.33%, then all of them declined to 12%-18% at the 6-month and 12-month follow-up periods. 65.43% of the patients resisted to both NVP and EFV but declined to 24%-27% at 6 months and 12 months.

CONCLUSIONHCV/HIV co-infected patients experienced virological failure of AIDS first-line therapy were ameliorated after changing to use second-line antiretroviral treatment for 6-months, but did not show constant positive effect at the 12-month end point.

Anti-HIV Agents ; pharmacology ; therapeutic use ; Antiretroviral Therapy, Highly Active ; CD4-Positive T-Lymphocytes ; China ; Coinfection ; Drug Resistance, Viral ; Follow-Up Studies ; HIV Infections ; complications ; drug therapy ; HIV-1 ; drug effects ; Hepatitis C ; complications ; drug therapy ; Humans ; Reverse Transcriptase Inhibitors ; pharmacology ; therapeutic use ; Treatment Outcome ; Viral Load

OBJECTIVETo investigate the effect of aloin on apoptosis of human gastric cancer cells and explore the molecular mechanism.

METHODSGastric cancer MKN-28 and HGC-27 cells were cultured routinely in 1640 medium supplemented with 10% fetal bovine serum and 10% non-essential amino acids (for HGC-27 cells) and treated with different concentrations of aloin for different durations. The cell viability, cell nuclear morphology, and apoptotic rate of the cells were detected using CCK-8 assay, DAPI staining and AnnexinV-FITC/PI, respectively; Western blotting was used to detect the expression levels of PARP, procaspase 3 and the phosphorylation of p38, ERK and JNK. The cells were treated with specific inhibitors of p38, ERK and JNK, and the inhibitory effects on these pathways were detected with Western blotting; DAPI staining was used to detect the effects of inhibitors on apoptosis of gastric cancer cells.

RESULTSAloin dose-dependently inhibited the viability and induced apoptosis of HGC-27 and MKN-28 cells. Alion treatment obvious enhanced the phosphorylation of p38 and JNK but decreased ERK phosphorylation in the cells. Blocking ERK activation with the ERK inhibitor obviously enhanced aloin-induced cell apoptosis, where inhibiting p38 and JNK activation partly reversed alion-induced apoptosis in the cells.

CONCLUSIONSAloin induces apoptosis of human gastric cancer cells by activating p38 and JNK signaling pathways and inhibiting ERK signaling pathway.