OBJECTIVETo investigate the effect of bufalin on nucleus-mitochondria localization of human telomerase reverse transcriptase(hTERT) by exploring its effect on proliferation and apoptosis in human esophageal squamous carcinoma EC9706 cells.

METHODSEC9706 cells were treated with bufalin at various concentrations, and then the cell growth inhibition of EC9706 cells was examined by CCK-8 assay and the 50% inhibitory concentration (IC(50)) was calculated.Cell cycle analysis was performed by flow cytometry with PI staining, and nucleus morphology of apoptosis were observed by fluorescence microscopy with Hoechst 33342 staining. The apoptotic index was measured by flow cytometry with Annexin V-FITC/PI double staining. hTERT subcellular localization and protein expression were determined by Western blotting and multiple immunofluorescence labling combined with laser confocal scanning microscopy.

RESULTSThe proliferation of EC 9706 cells was significantly inhibited by bufalin along with the increase of processing time and concentrations (p<0.01). After the EC9706 cells were exposed to 100 nmol/L bufalin,the number of cells gradually decreased in G(1) phase and increased in S and G(2)/M phases(p<0.05). The typical nucleus morphological changes of apoptosis were observed and the apoptotic index was increased(p<0.01). The expression of hTERT decreased in nucleus but increased in mitochondria(p<0.05).

CONCLUSIONSBufalin can inhibit the proliferation of human esophageal squamous carcinoma EC9706 cells in a time- and dose-dependent manner. It can arrest cell cycle in S and G(2)/M phases and induce the apoptosis of EC 9706 cells. hTERT is localized in both nucleus and mitochondria,and can be partially translocated from nucleus to mitochondria during the bufalin-induced apoptosis.

Apoptosis ; drug effects ; Bufanolides ; pharmacology ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Esophageal Neoplasms ; metabolism ; pathology ; Humans ; Telomerase ; metabolism

Objective To understand the status quo of health resource allocation of medical institutions in poverty-stricken areas, and to provide decision-making basis for rationalizing health and poverty alleviation policies and improving the overall service capacity of medical institutions in poverty-stricken areas. Me thods The overall institutions, bed capacity and staffs in medical institutions in 680 poor counties were analyzed. Re s ults The proportion of government health expenditure in 14 concentrated areas was lower than 15％. The largest number of health institutions was 349 in the Dabie Mountains and 70 in Tibet, and the number of beds was lower than the national average level of 5.11.The largest number of health technical staff for 1 000 people of the four provinces is 4.42 people, the smallest number is 2.72 in Wumeng mountain area;the registered nurses (number) for 1 000 people is up to 1.56 people in the Luo Xia mountain area, the lowest Tibet, only 0.39 people. Thousands of population practice (assistant) physician number of Tibetans is up to 2.98 people, the lowest is 1.07 for the Xinjiang Southern Xinjiang three states; health care than the lowest in Tibet 1:0.54. Conclus ion At present, China's centralized contiguous poverty-stricken areas of county-level medical institutions is extremely short of resources, and the health resource allocation is uneven.

Chinese agarwood is formed in the aromatic resinous wood formed in Aquilaria sinensis (Lour.) Gilg (botanical family: Thymelaeaceae). Only when suffering stress of wound, etc, can A. sinensis produce sesquiterpenes etc. compounds of agarwood around wounds. However, little is known about how wound induced the biosynthesis pathway of sesquiterpenes. To reveal the molecular mechanism of wound-induced agarwood formation, RNA sequencing (RNA-seq) technology was used to investigate the profile of gene expression in A. sinensis treated by mechanical wounding and elucidate its functional gene. A total of 40,295 ESTs with an average read length of 305 bp were generated and 22 095 unigenes were formed by initial gene splicing. 61.6% of these unigenes (13 611) were annotated using BLAST searches against the SwissProt, KEGG, Nr and Nt databases. Twenty-six unigenes (encoding 7 enzymes) were found to be involved in sesquiterpene of agarwood biosynthesis by bioinformatic tools of Gene Ontology and KEGG. Novel genes that are potentially involved in sesquiterpenes biosynthesis were identified in A. sinensis, providing data for further sesquiterpenes biosynthesis pathway by molecular methods and the EST data establish a foundation for future studies in the molecular mechanisms of wound-induce agarwood formation in A. sinensis.
Drugs, Chinese Herbal ; chemistry ; metabolism ; Expressed Sequence Tags ; Genes, Plant ; genetics ; Plants, Medicinal ; chemistry ; genetics ; Sequence Analysis, RNA ; Sesquiterpenes ; chemistry ; metabolism ; Stress, Physiological ; genetics ; Thymelaeaceae ; chemistry ; genetics ; Transcriptome ; genetics ; Wood ; genetics ; metabolism

AIM To investigate the effects of Moluodan Dami Pills on chronic atrophic gastritis(CAG)rats and their mechanism.METHODS The rat models were randomly divided into the model group,the low-dose group and high-dose Moluodan Dami Pills groups(2.43 g/kg and 4.86 g/kg),and vitamin A group(0.32 g/kg),following the 16 weeks successful induction of CAG by five-factor modeling method,in contrast to another 10 normal rats of the control group.After 8 weeks corresponding administration,the rats of each group had their general physiological status and pH value of gastric juice assessed;their pathological changes of gastric mucosa observed by naked eyes combined with HE staining;their changes of gastrin-secreting cells(G cells)and somatostatin-secreting cells(D cells)in gastric mucosa observed by immunohistochemistry;and their serum levels of pepsinogen Ⅰ/pepsinogen Ⅱ(PG Ⅰ/PG Ⅱ)ratio,TNF-α and IL-6 detected by ELISA.RESULTS Compared with the model group,the groups intervened with Moluodan Damei Pills and vitamin A displayed lower pH values of gastric juice(P＜0.05),improved pathological changes of gastric mucosa,increased G and D cells counts(P＜0.05,P＜0.01),increased ratio of serum PGⅠ/PGⅡ,and decreased levels of IL-6 and TNF-α(P＜0.05,P＜0.01).CONCLUSION Moluodan Dami Pills can effectively improve the symptoms of CAG rats through its influence on the number and secretion abilityof G and D cells,the levels of serum PG Ⅰ/PG Ⅱ ratio and inflammatory factors.

Objective To explore the expression characteristics of cytochrome P450(CYP)candidate genes(CYP6M3, CYP6Y1,CYP6P5,CYP4H14,CYP4G17,CYP12F16)in Anopheles sinensis.Methods The samples were collected from dif-ferent developmental stages(eggs,larvae,pupae and adult mosquitoes(females and males)),and different tissues(salivary glands,malpighian tubes,midguts,ovaries,and fat bodies)of An.sinensis and the female adult mosquitoes exposed by differ-ent insecticide doses(0,1.25,3.75,6.25,12.5μg/bottle)and time points(0,5,15,30,60 minutes),then the total RNA was extracted. Quantitative reverse transcription PCR(qPCR)assay was used to analyze the relative expressions of six CYP genes in An.sinensis at different developmental stages,tissues and different insecticide exposure doses and time.Results The expressions of CYP6M3 and CYP6Y1 in the male adult mosquitoes were the highest,the expression of CYP6M3 gene in the males was 35.1 times higher than that in the females,the expression of CYP6Y1 in the males was 61.4 times higher than that in the females;the expression level of CYP4H14 in the larvae was the lowest,the expression of CYP4H14 in the females was 22.5 times higher than that in the fourth instar larvae.The expressions of candidate CYP genes in different tissues of An.sinensis were significantly different,the expression of CYP6M3 in the malpighian tubule was 38.9 times higher than that in the ovary,the ex-pression of CYP6Y1 in the fatbody was 9.1 times higher than that in the ovary,the expression of CYP4G17 was 4.6 times higher than that in the ovary,and the expression of CYP12F16 was 4.4 times higher thanthat in the ovary.The exposure to different in-secticide doses and time showed some induction effects in the expressions of candidate CYP genes,which affected the expres-sions of candidate CYP genes in An.sinensis.Conclusion The expression of candidate CYP genes varies significantly in differ-ent developmental stages and various tissues of An.sinensis,and exposure to deltamethrin at various doses and time points af-fects CYP genes expression in An.sinensis.

Objective:To explore the active components， targets， and signaling pathways responsible for Bushen Zhuyun prescription in treating the recurrent spontaneous abortion （RSA） based on network pharmacology and uncover its potential mechanism by molecular docking and in vitro cell experiments. Method:The active components of Bushen Zhuyun prescription were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）， Traditional Chinese Medicine Integrated Database （TCMID） and the published articles， followed by the prediction of drug action targets based on such platforms as DrugBank and SwissTargetPrediction. GeneCards and Online Mendelian Inheritance in Man （OMIM） were searched to obtain the RSA targets， which were then intersected with the targets of Bushen Zhuyun Decoction. Following the plotting of Bushen Zhuyun prescription-compound-target-RSA network by Cytoscape 3.7.1， the protein-protein interaction （PPI） network was then constructed with STRING for screening the core network. The resulting common targets were then subjected to Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis using R software. Autodock Vina 1.1.2 was used for molecular docking. The activation of phosphatidylinositol 3-kinase/protein kinase B （PI3K/AKT） signaling pathway by Bushen Zhuyun prescription was verified in human umbilical vein endothelial cells （HUVEC） in vitro. Result:It was found that 49 potential active components of Bushen Zhuyun prescription might act on 133 RSA targets. GO enrichment analysis yielded 470 biological processes， with angiogenesis， vascular development， cellular proliferation， and oxidative activity mainly involved. KEGG enrichment analysis revealed 103 signaling pathways （P<0.05）， and the PI3K/AKT signaling pathway， advanced glycation end product （AGE）/receptor for advanced glycation end product （RAGE） signaling pathway， and tumor necrosis factor （TNF） signaling pathway were the main ones. As indicated by molecular docking， the Vina scores of the main active component kaempferol with AKT1 and vascular endothelial growth factor A （VEGFA） were the lowest and similar. It was confirmed in vitro cell experiments that Bushen Zhuyun prescription activated the PI3K/AKT signaling pathway and up-regulated the expression of VEGFA and downstream AKT protein to promote angiogenesis. Conclusion:Bushen Zhuyun prescription promotes angiogenesis at the maternal-fetal interface by regulating angiogenesis and cellular proliferation， activating the PI3K/AKT pathway， and up-regulated the VEGFA expression， which is beneficial to the formation of placenta in early pregnancy and the maintenance of early pregnancy. This study has provided ideas for new drug development.

Objective:To evaluate the effect of Kinesio taping on chronic nonspecific low back pain (CNLBP). Methods:The Cochrane Library, PubMed, Web of Science, CNKI, CBM, VIP, and Wanfang Data were searched for the randomized controlled trials (RCT) about the effect of Kinesio taping on CNLBP from establishment to January, 2019. The included studies were evaluated according to the method recommended by the Cochrane Collaboration. RevMan 5.3 software was used to analyze the extracted data. Results:Finally, nine RCTs involving 545 patients were included. Meta-analysis showed that the effect of Kinesio taping was significantly different in the improvement of pain compared with the non-stimulated group (MD = -0.76, 95%CI: -1.43 to -0.08, P = 0.03), the difference might be significant compared with the sham stimulation group (MD = -1.10, 95%CI: -2.22 to 0.02, P = 0.05); For improving dysfunction, the Oswestry Disability Index (ODI) scores were better in the Kinesio taping group than in the non-stimulation group (MD = -6.02, 95%CI: -8.63 to -3.41, P < 0.001) and the sham stimulation group (MD = -4.11, 95%CI: -5.82 to -2.41, P < 0.001), however, their was no significant difference in Roland Morris Disability Questionnaire (RMDQ) score between the Kinesio taping group and the non-stimulated group (MD = 0.69, 95%CI: -2.35 to 3.74, P = 0.66), and between the Kinesio taping group and the sham stimulation group (MD = -0.17, 95%CI: -1.43 to 1.08, P = 0.78). Conclusion:For the patients with CNLBP, the intervention of Kinesio taping could alleviate pain and improve function.

To understand the quality of production of blood slides and laboratory personnel’s ability to microscopically examine Plasmodium at Jiangsu Province clinic inspection stations at all levels, so as to provide the evidence for the construction of malaria diagnostic laboratories and the evaluation of malaria elimination effect.After the reexamination of the negative blood slides of the examination stations of the malaria center in each county (district), the negative blood slides of the outpatient inspection station in the district and all the positive blood slides of the direct report of the network from 2011 to 2017, the staining and cleanliness of the blood smears, microscopic examination results and the coincidence rate of positive blood species were comprehensively analyzed.A total of 19 109 checked blood samples were examined. The qualified rates of blood preparation, dyeing and cleanliness were 87.5%, 89.3% and 87.6%, respectively. The quality of reexamined blood slides in different years showed statistically significant difference (all P < 0.05), by using chi-square trend test, and there was a trend of increasing year by year. There was a significant difference in the quality of the reexamined blood slides among the different cities with districts (P < 0.05). In addition, there were 1 860 blood samples from 2012 to 2017, and the total coincidence rate was 87.4% and the coincidence rate of positive blood smear examinations was 98.9%. The species coincidence rates of Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, Plasmodium ovale and mixed infection were 95.4%, 33.3%, 56.0%, 76.5% and 9.1%, respectively (P < 0.05). The total coincidence rates of blood smear reexaminations in different years were 85.0%, 90.1%, 86.0%, 85.3%, 90.0% and 87.6% from 2012 to 2017, respectively, and the difference was not statistically significant (P > 0.05).The quality of blood slides at the primary clinic inspection stations in Jiangsu Province conforms to the standard established by the provincial malaria reference laboratory; however, the ability of Plasmodium species identification is insufficient and needs to be improved.

Objective To analyze the epidemic situation of malaria in Jiangsu Province in 2018, so as to provide scientific evidence for formulating post-elimination malaria surveillance schemes and technical measures in Jiangsu Province. Methods The malaria case report cards, epidemiological individual investigation forms of malaria cases and foci data were collected from Jiangsu Province in 2018, and the epidemic situation of malaria was descriptively analyzed. Results A total of 243 malaria cases were reported in Jiangsu Province in 2018, which increased by 1.67% in relative to in 2017 (239 cases), and these cases included 171 cases with falciparum malaria, 14 cases with vivax malaria, 15 cases with quartan malaria, 42 cases with ovale malaria and a case with mixed infection of P. vivax and P. ovale. All cases were overseas imported, and no local secondary cases were found. The malaria cases were predominantly workers (76.54%). Nantong City (48 cases), Yangzhou City (33 cases) and Taizhou City (22 cases) were the most 3 cities with the largest number of malaria cases across Jiangsu Province. The malaria infections predominantly occurred in African areas (96.30%), and the other 9 cases had infections in Asia (8 cases) and Central America (1 case). There were 125 cases (51.44%) and 91 cases (37.45%) with definitive diagnosis at the day of admission and within 1 to 3 days post-admission, respectively. The percentages of definitive diagnosis at initial diagnosis were 48.27%, 88.76% and 97.30% at township-, county- and city-level medical institutions, respectively, and the percentage of definitive diagnosis at initial diagnosis was significantly lower in township-level medical institutions than in county- (χ² = 21.47, P < 0.01) and city-level medical institutions (χ² = 32.86, P < 0.01). Conclusions There are no local malaria cases in Jiangsu Province; however, the number of overseas imported malaria cases remains high in China. In the future, improving the post-elimination malaria surveillance system, enhancing the awareness of malaria prevention and control knowledge among high-risk populations, increasing the diagnostic capability of malaria in medical institutions, and improving the management of imported malaria cases should be performed to consolidate the achievements of malaria elimination.

Objective To analyze the epidemic situation of malaria in Jiangsu Province in 2019, so as to provide the scientific basis for the development of the strategy for the prevention of re-introduction of imported malaria. Methods The malaria case report information, epidemiological case investigation information, epidemic foci investigation and management report in Jiangsu Province in 2019 were collected, and all epidemiological data were descriptively analyzed. Results A total of 244 malaria cases were reported in Jiangsu Province in 2019, and all cases were laboratory-confirmed overseas imported cases, including 4 cases with vivax malaria, 206 cases with falciparum malaria, 12 cases with malariae malaria and 22 cases with ovale malaria. In 2019, there were 12 malaria cases progressing into severe cases in Jiangsu Province, with one death. Nanjing, Nantong, Lianyungang, Taizhou and Changzhou cities contributed the largest number of malaria cases in 2019, with the number of malaria cases accounting for 59.84% of total cases in Jiangsu Province. The infections occurred in Papua New Guinea (2 cases), Pakistan (1 case) and 27 African countries (241 cases), including Angola, the Democratic Republic of the Congo, Nigeria, Equatorial Guinea, Cote d'Ivoire and so on. There were 77 cases (31.55%) with a visit to doctor on the day of onset, and 146 cases (59.84%) within 1 to 3 days after onset. In addition, there were 149 cases (61.06%) with definitive diagnosis at the first visit and 77 cases (31.55%) diagnosed within 1 to 3 days after the visit, and the mean duration from the visit to definitive diagnosis was (0.80 ± 1.59) d, which significantly shortened as compared to that (1.34 d ± 2.59 d) in 2018 (U = 2.53, P < 0.05). Conclusions Intensifying the surveillance and management of imported malaria and improving the diagnostic capability of imported malaria and the treatment of severe malaria cases are required to consolidate the achievements of malaria elimination in Jiangsu Province.