2.Role of solasodine hydrochloride in AS2O3 induced HeLa cells apoptosis as well as its effect on cell telomerase activity in vitro
Jin-Xia, AI ; Liang, LIU ; Ping, WANG
Chinese Journal of Endemiology 2011;30(3):279-283
Objective To study whether solasodine hydrochloride (SBHL) could enhance the effect of arsenic trioxide in inducing apoptosis and affecting telomerase activity in cervical cancer HeLa cells. Methods Using cell culture methods, cervical cancer HeLa cells were cultured in vitro. The optimal concentration of SBHL was determined by MTT method from 0, 10, 20, 40, 80, 160, to 320 μmol/L. HeLa cells were grown in improved RPMI1640 supplemented respectively with arsenic trioxide(5 μmol/L As2O3), As2O3(5 μmol/L)+ SBHL( 40 μmol/L) and none (control group). The growth morphology of HeLa cells was observed under phase contrast microscopy after culture for 24, 48, and 72 h. Apoptosis of HeLa cells was determined under transmission electronic microscopy. The method of MTT was used to study the cell survival percentage. The technique of flow cytometry was used to measure cell cycle and cell apoptosis percentage. The method of tartrate-resistant acid phosphatase-enzyme linked immunosorbent assay (TRAP-ELISA) was used to determine telomerase activity of HeLa cells. Results Under phase contrast microscopy, in control group HeLa cells were round, densely packed; in As2O3 group the numbers of the cells were less, cell spacing increased; in As2O3 + SBHL group the cells shrinked significantly, nuclear fragmented as a petal-like, gap became larger. Under transmission electronic microscopy, there were rich microvillus on the cell surface in control group, cell intervals clear, immature connections, and the intervals did not close. The structure of the mitochondria in the cytoplasm was integrated. Most of the chromatin in the nucleus were, euchromatin and characteristics of apoptosis with heterochromatin increased and the chromatin condensed into masses, on the boundary of nuclear membrane. The microvillud on the cell surface were ruptured and decreased in As2O3 + SBHL group. The chromatin condensed into masses. The formation of apoptotic bodies was observed. The difference was statistically significant between groups in cell survival percentage at 24, 48, 72h(x2 = 10.39 , 13.88 , 17.21,respectively, all P < 0.05). Cell survival percentage in SBHL + As2O3 group (52.80%) was significantly less than that of As2O3 group(77.51%, x2 = 9.29, P < 0.05) at 72 h. In cell cycles, the difference was statistically significant between groups in C1 phase and S phase(F = 7.46,22.14, all P < 0.05), respectively. Compared with , control group[ (41.57 ± 1.56)%, (50.45 ± 2.37)%], cell percentages in S phase in As2O3 + SBHL group[(20.06 ± 4.98)%] and As2O3 group[(27.10 ± 5.32)%] were decreased(P< 0.05 or < 0.01), while cell percentage in C1 phase was increased[(58.70 ± 5.18)%, (69.67 ± 4.17)%, P< 0.05 or < 0.01]. The difference was statistically significant between groups in apoptotic percentage of HeLa cells (F = 4.01, P < 0.05). Compared with control group[ (1.18 ± 1.40)%], apoptosis percentage was significantly increased in As2O3 + SBHL group and As2O3 group [(21.08± 1.22)%, (6.04±2.53)%, P< 0.05 or < 0.01], respectively, and As2O3 + SBHL group was higher than As2O3 group(P < 0.01). The difference was statistically significant between groups in telomerase activity (F = 21.28, P< 0.05). Telomerase activity was inhibited in As2O3 group(1.214 ± 0.621) and As2O3A + SBHL group(0.865 ± 0.284) compared to control group (2.107 ± 0.057, all P < 0.05), and telomerase activity in As2O3 + SBHL group was lower than that of As2O3 group (P < 0.05). Conclusions SBHL enhances the effect of As2O3 in inducing apoptosis in HeLa cells, which is related to its inhibiting telomerase activity in HeLa cells.
3. Development and Evaluation of DNA Detection Kit for Fetus Cervi
Chinese Pharmaceutical Journal 2018;53(4):306-311
OBJECTIVE: To develop a DNA detection kit for identification of Fetus Cervi, evaluate the quality indexes including specificity, stability, sensitivity and repeatability, and inspect the qualities of commercial Fetus Cervi samples. METHODS: The Fetus Cervi DNA test kit was developed and modified using the classical DNA extraction and PCR identification method. The genomic DNAs of Fetus Cervi and counterfeit goods were extracted by the kit and PCR technique was performed to identify the authenticity. The purity of DNA was detected by UV spectrophotometer. Finally, commercially available Fetus Cervi samples were verified. RESULTS: The kit was proved to be effective after 20 times of repeated frozen-thawing and it could be stored at - 20℃ for 1 year. The DNA could be detected when the primary solution was diluted by 200 times. The specificity test confirmed that the 15 samples were authentic Fetus Cervi, and 7 were counterfeit. The specificity of the kit was 100%. The repetitive experiments showed that the average intra-assay CV% and inter-assay CV% of the kit were 2.32% and 2.56%, respectively. CONCLUSION: The amount and purity of the nucleic acids extracted by the DNA detection kit can meet the requirement for identification of Fetus Cervi, and the kit has good specificity, high sensitivity and good stability, so it is suitable for the rapid detection of Fetus Cervi.
4. Application of Mitochondrial DNA in the Identification of Fetus Cervi
Chinese Pharmaceutical Journal 2017;52(18):1589-1593
OBJECTIVE: To explore a practical method for isolation of mitochondrial DNA from Fetus Cervi and set up an accurate method to distinguish Fetus Cervi from adulterants. METHODS: The improved SDS alkaline lysis and salt-outing METHODS were used to extract mtDNA and genomics DNA from Fetus Cervi and fetus from adulterant animals respectively. A set of primers were designed by bioinformatics to establish PCR system and reaction which could be used to identify Fetus Cervi species. RESULTS: The mtDNA extracted from Fetus Cervi by improved SDS alkaline lysis method had high content and purity, which met the PCR response requirements. There was a pair of primers which could distinguish Fetus Cervi from adulterants. CONCLUSION: Extraction of mtDNA by improved SDS alkaline lysis method is practical and accurate. The distinctive PCR can distinguish Fetus Cervi from adulterants and counterfeit animal fetus and it is accurate and reliable.
5.Effect of comprehensive prevention and control of soil-transmitted nemato-diasis in Runzhou District of Zhenjiang City
Ai XIA ; Hengye TAO ; Yaming ZHAO ; Jun JIANG ; Jin WANG ; Jin MEI
Chinese Journal of Schistosomiasis Control 2014;(6):665-668
Objective To evaluate the effect of comprehensive prevention and control of soil?transmitted nematodiasis in Runzhou District Zhenjiang City Jiangsu Province from 1997 to 2012. Methods The comprehensive prevention and control measures included the helminthicide health education improvement of water supplier and harmless toilets and these mea?sures were implemented continuously. At the same time the infection rates of soil?transmitted nematodes the local economic in?dicators and the coverage rates of tap water and harmless toilets were surveyed. Results The mass chemotherapy was per?formed for 202 100 person?times and the diagnosed chemotherapy was performed for 2 006 person?times in Runzhou District from 1997 to 2012. The awareness rates of health knowledge were 57.18%in 1997 and 95.62%in 2012. The coverage rates of tap water and harmless toilets were 10.14%and 10.21%in 1997 and 100.0%and 90.38%in 2012 respectively. There were negative correlations between the awareness rate of per capita GDP per capita net income coverage rates of tap water cover?age rates of harmless toilets health knowledge and the infection rate of soil?transmitted nematodes respectively rper capita GDP=-0.526 P 0.05 rper capita net income=-0.671 P 0.01 rcoverage rates of tap water=-0.936 P 0.01 rcoverage rates of harmles toilets=-0.922 P 0.01 rawarenes=-0.774 P 0.01 . The statistical analysis showed that the infection rate of soil?transmitted nematodes had a downward trend as an expo?nential curve in Runzhou District from 1997 to 2012 y=42.031 7e?0.357 6x R2=0.803 6 F=57.28 P=0.000 . The infection rate of degradation by an annual rate was 29.18%. The infection rate in farmers was significantly higher than that in students χ2=17.998 P 0.01 . There was no significant difference between men and women in the infection rate of soil?transmitted nema?todes χ2=3.627 P=0.057 . Conclusion The comprehensive prevention and control measures and the development of so?cial economy contribute to the steady decline of soil?transmitted nematode infections.
6.Three cases of enterovirus 71 infection with pulmonary edema or pulmonary hemorrhage as the early clinical manifestation
Shi-Jun HE ; Xia WANG ; Xiao-Qun ZHENG ; Chuan-Xia WANG ; Ai-Rong HUANG ; Yi-Mei JIN ; Hao-Mei YANG ; Ai-Hua ZHOU
Chinese Journal of Pediatrics 2008;46(z1):49-53
Objective To investigate the clinical features of the enterovirus 71 ( EV71 ) infection complicated with pulmonary edema or pulmonary hemorrhage as a fulminant and often fatal illness.Methods The medical records of three cases with EV71 infection were retrospectively reviewed for clinical manifestations, laboratory data, medications, and outcome.Results All the cases were infants and died of the infection. These infants had no skin or mucosal lesions, however, they had sudden onset of cyanosis and tachypnea 1 to 2 days after the onset of the febrile disease with vomiting. All these 3 cases were misdiagnosed and were treated for shock on admission. Pulmonary hemorrhage was not considered in any of the cases on admission. All the cases received tracheal intubation when foamy secretions were discharged from the mouth and nose of the patients and notable cyanosis occurred. After intubation, pink foamy fluid flew out from the endotracheal tube in all the 3 cases. The patients had hyperglycemia and limb weakness, two had tachycardia, and hypertension was found in one case. Chest X-ray showed bilateral or unilateral widespread air space opacity, but the cardiac size and shape were normal. All the patients had leukocytosis. Enterovirus 71 infection was confirmed by detection of specific nucleic acid sequences of the virus from throat swab and tracheal secretions samples and in one case in cerebrospinal fluid.Conclusions Pulmonary edema or pulmonary hemorrhage occurred in the 3 cases with EV71 infection. The initial presentation was nonspecific with fever and vomiting, and sudden appearance of cyanosis, tachypnea, tachycardia, hypertension or hypotension, limb weakness, which may suggest pulmonary edema or hemorrhage. Excessive fluid resuscitation may deteriorate the illness, on the contrary, fluid restriction and inotropic agents, and early intubation with positive pressure mechanical ventilation may be the proper treatment.
7.The Mutagenic Effect on PHB Accumulation of Acidiphilium cryptum DX1-1
Ai-Ling XU ; Shuai ZHANG ; Yan-Fei ZHANG ; Li LI ; Yu YANG ; Jin-Lan XIA ;
Microbiology 2008;0(10):-
The strain Acidiphilium cryptum DX1-1 producing PHB was irradiated respectively by UV and Co60 to raise PHB production. The results indicated that the effect of UV better than using Co60. One strain of the UV mutagenized called UV60-3 has the highest PHB production yield, showing final PHB concentra- tion of 28.56 g/L, 1.45 times higher than that of original strain. FT-IR spectroscopy analysis shows that the polymers obtained from the strain DX1-1 have the same IR spectra of standard PHB. Further research about the best appropriate C/N ratio of the mutant was done. The optimum ratio of C/N was about 3.76, the final PHB concentration reaches to 30.57 g/L.
8.Analysis on family function of community residents and its influential factors
Jin-Bing BAI ; Li-Ping YU ; Yan-Qun LIU ; Xia LIU ; Ai-Ling WANG
Chinese Journal of Modern Nursing 2009;15(1):33-35
Objective To explore the current status of the family function and the influential factors in the community, Methods A questionnaire survey including the general data questionnaire and APGAR scale developed by Smikstein was conducted in 94 families of three communities of Wuhan City. Results 46.8% of the surveyed family showed good function, 51. 1% showed moderate dysfunction and 2.1% in severe dysfunction. The multinomial regression revealed that education level and whether the family members had chronic diseases had influence to the family function level (P<0.05). Conclusions More than 50% of the families exist moderate or severe family dysfunction. The higher education level, the better family function; meanwhile compared with the family whose members experience chronic diseases, the family without experiencing chronic dieseases shows much better family function.
9.Status of health literacy of undergraduate nursing students in one university of Lanzhou City
Jin XIA ; Hui-Lan LI ; Ai-Ling QIN
Chinese Journal of Modern Nursing 2013;19(3):331-332
Objective To understand the status of health literacy among undergraduate nursing students in one University of Lanzhou City so as to provide the reference evidence for health education for undergraduate nursing students.Methods Independent design questionnaire based on " Health literacy questionnaire about Chinese citizens in 2008",and random sampling 243 undergraduate nursing students of one University of Lanzhou City were investigated.Results Among students,the awareness rates of health literacy,basic knowledge and concept of health,healthy life style and behavior,and the basic skill respectively were 41.95%,45.49%.29.24% and 60.43%.No significant difference was found in the awareness rates among students with different sexual (x2 =1.83,2.23,1.01,0.48,respectively ; P > 0.05),while significant difference was found in the freshman and others students (x2 =59.17,33.33,21.99,8.29,respectively;P < 0.01),town students and rural students (x2 =25.69,10.30,10.58,6.89,respectively; P < 0.01).Conclusions Awareness rate of the health literacy of undergraduate nursing students of the university is higher than that of our country dweller.
10.Clinical analysis of community-acquired Pseudomonas aeruginosa septic shock.
Shi-jun HE ; Yi-mei JIN ; Ai-rong HUANG ; Chuan-xia WANG ; Ai-hua ZHOU ; Xia WANG ; Xiao-ou SHAN
Chinese Journal of Pediatrics 2008;46(5):333-339
OBJECTIVEThis study sought to analyze the clinical manifestations and intervention of fulminant septic shock in community-acquired Pseudomonas aeruginosa septicemia.
METHODSWe retrospectively reviewed the medical records for diagnosis, antibiotic therapy, clinical course of septic shock, respiratory support, laboratory data etc.
RESULTSEight of nine cases with P. aeruginosa septic shock died. Fever (nine cases) and cough (three cases) or diarrhea (3 cases) were the 2 most common initial symptoms, three cases developed skin gangrenosum later. Pseudomonas aeruginosa infection was not considered in any of the cases before death or blood culture showed positive result. Only 3 cases were initially treated with susceptible antibiotic regimen but no anti pseudomonas combination therapy was applied, susceptible antibiotic monotherapy was applied in 7 cases after transfer to the ICU. The mean latency of shock occurrence was 5.1 hours (range 0 to 21 hours) after admission, the mean duration from the occurrence of shock to death was 13.8 hours (range, 1 - 32 hours). All the patients were transfer red to ICU for shock, the appropriate resuscitation of shock patients was delayed by 49.3 minutes (range 25 - 80 minutes) by transfer. Only two cases were diagnosed and treated for shock on admission; after transferred to ICU, only 5 patients were diagnosed as having shock, and only 3 received anti-shock treatment. Eight of the patients died of persistent shock. In 6 patients who died, mechanical ventilation was not applied until cardiac arrest occurred. All the patients had hypoalbuminaemia, elevated serum C-reactive protein concentration, leukopenia and 6 cases had DIC.
CONCLUSIONThe initial presentation of the cases with community-acquired Pseudomonas aeruginosa septicemia was nonspecific with fever and cough or diarrhea. Clinicians often underestimated the severity of the infection, few patients received effective antimicrobial therapy. The authors suggest that an anti-pseudomonas antibiotic should be included in the initial empiric antibiotic regimen to cover P. aeruginosa high-risk patients; the front-line clinician should be educated for early recognition and aggressive resuscitation of P infection. aeruginosa septicemia.
Adolescent ; Child, Preschool ; Community-Acquired Infections ; Female ; Humans ; Infant ; Male ; Pseudomonas Infections ; Pseudomonas aeruginosa ; Retrospective Studies ; Shock, Septic ; microbiology