1.Effect of the transcription factor SOX3 on ovarian granulosa cell proliferation and estradiol secretion
Rui CAI ; Hao ZHANG ; Zhuang LIU ; Yuanhua CHEN ; Fenfen XIE ; Qiang HONG
Acta Universitatis Medicinalis Anhui 2024;59(3):371-376,383
Objective To study the effect of sex-determining region Y-frame protein 3(SOX3)on proliferation and estradiol secretion in human ovarian granulosa cells(KGN cell line).Methods The gene sequence of human SOX3(NM_005634.3)was searched in Gene-Bank,an NCBI database,and the target gene SOX3 was amplified by PCR,which was cloned into lentiviral vector pLV-EF1a-GFP-2A-Puro to obtain the overexpression lentiviral re-combinant plasmid pLV-EF1a-GFP-2A-Puro-SOX3;the correctly sequenced overexpressed lentiviral recombinant plasmid as well as packaging plasmids(pGag/Pol,pRev,pVSV-G)were co-transfected into human embryonic kidney cell line(HEK 293T)cells(pLV-SOX3 group),and pLV-EF1a-GFP-2A-Puro and packaging plasmids(pGag/Pol,pRev,pVSV-G)were co-transfected into HEK 293T cells(pLV-NC group),the lentiviral particles of both groups were collected and the titers of the viruses were measured after 48 h of transfection,the lentiviruses of the two groups were infected into KGN cells,and the stably expressed cell lines were obtained after puromycin screening for 2 weeks;real-time fluorescence quantitative PCR(RT-qPCR)and Western blot were used to detect the SOX3 mRNA and protein levels in the two groups;CCK-8 assay was used to detect the proliferative ability of the cells in the two groups;ELISA was used to determine the concentration of estradiol in the two groups.Results The identification of PCR products and sequencing results showed that the SOX3 gene fragment was amplified successfully,and the enzyme digestion and sequencing results indicated that the construction of overexpression lentiviral recombinant plasmid was completed;green fluorescence could be detected after lentiviral infection of HEK 293T cells,which indicated that lentiviral packaging was successful;the lentivirus was screened by puromycin after lentiviral infection of KGN cells,and the cells were observed to express green fluorescence under the fluorescence microscope;RT-qPCR and Western blot assays both showed that the expression level of SOX3 in the pLV-SOX3 group was significantly higher than that in the pLV-NC group(P<0.05).CCK-8 assay results showed that the proliferation ability of the cells in the pLV-SOX3 group significantly increased compared with that in the pLV-NC group(P<0.01).ELISA results showed that estradiol concentration was elevated in the pLV-SOX3 group com-pared with the pLV-NC group(P<0.05).Conclusion Overexpression of the transcription factor SOX3 can pro-mote the proliferation and estradiol secretion of human ovarian granulosa cells KGN.
2.The Effect of TCM Syndrome and Th17/Treg Cells Immunomodulatory of Yiqi Jiedu Compound on Patients with Myasthenia Gravis
Yalin LIU ; Yuanhua WU ; Qing GUO ; Youde CAI ; Shanshan WANG ; Chen LI ; Qiansong HE
World Science and Technology-Modernization of Traditional Chinese Medicine 2023;25(6):2203-2208
Objective To observe the effects of Yiqi Jiedu Compound on TCM syndrome score and immune regulation of Th17/Treg cells in peripheral blood of patients with myasthenia gravis.Methods 63 patients with myasthenia gravis were randomly divided into the control group(n=31)and the treatment group(n=32).The control group was given the basic treatment of pyridostigmine bromide and glucocorticoid and so on,and the treatment group was given Yiqi Jiedu Compound based on the control group.The two groups were treated for 12 weeks.Clinical efficacy was observed,TCM syndrome scores were statistically analyzed,and the expression levels were detected,which Th17/Treg cells,Foxp3/RORγ t gene and IL-6 and IL-17 cytokines in peripheral blood of patients in both groups before and after treatment.Results After 12 weeks of treatment,compared with the control group,the TCM syndrome score of patients in the treatment group was significantly decreased(P<0.05),and the expression of Foxp3 and Treg genes in peripheral blood was promoted(P<0.05),and the expression of Th17 and Treg cells and IL-6 and IL-17 cytokines in peripheral blood was decreased(P<0.05).Conclusion Yiqi Jiedu Compound may regulate the immune balance of Th17/Treg cells by affecting the differentiation of Th17 and Treg cells in peripheral blood and the expression levels of transcription factors RORγ t and Foxp3 genes and related cytokines,thus achieving the effect of myasthenia gravis treatment.
3.Trihalomethanes and haloacetamides as disinfection by-products in different types of drinking water of Wuxi
Run ZHOU ; Li YANG ; Yukang WU ; Zhifei XU ; Yuanhua MENG ; Xinliang DING ; Xiaofeng CHEN
Journal of Environmental and Occupational Medicine 2023;40(4):456-461
Background The concentrations of disinfection by-products (DBPs) are varied by different water sources, disinfectants, or treatment processes in Wuxi, and the associated health risks are also different. Objective To understand the levels of trihalomethanes (THMs) and haloacetamides (HAcAms) in drinking water in Wuxi, and their variations by water sources, seasons, disinfectants or treatment processes, aiming to provide technical support for ensuring the safety of drinking water. Methods In dry period (December 2019) and wet period (July 2020), the finished water and tap water (from the beginning, middle, and end of the drinking water distribution network) from 12 centralized water treatment plants in Wuxi were collected to detect the concentrations of THMs and HAcAms in water samples. A purge and trap-gas chromatography-mass spectrometry method was applied to detect trichloromethane (TCM), bromodichloromethane (BDCM), dibromochloromethane (DBCM), and tribromomethane (TBM), and a solid-phase extraction-gas chromatography-mass spectrometry method to detect dichloroacetamide (DCAcAm), trichloroacetamide (TCAcAm), bromochloroacetamide (BCAcAm), dibromoacetamide (DBAcAm), bromodichloroacetamide (BDCAcAm), dibromochloroacetamide (DBCAcAm), and tribromoacetamide (TBAcAm). Analyses and comparisons were made on the concentrations of THMs and HAcAms in drinking water by water sources (the Yangtze River/the Taihu Lake/reservoir), wet/dry seasons, disinfection methods (liquid chlorine/sodium hypochlorite), and treatment processes (conventional treatment/conventional+advanced treatment). Results A total of 96 drinking water samples were collected in Wuxi. THMs were positive in all the water samples (100%), with concentration ranging from 1.027 to 40.225 μg·L−1 and the M (P25, P75) concentration being 24.782 (17.784, 30.932) μg·L−1. None of the 4 THMs exceeded the standard limit of the Standards for drinking water quality (GB 5749-2022 ), and the order of the 4 THMs concentrations from high to low was TCM > BDCM > DBCM > TBM. Five of the 7 HAcAms were detected, the total concentration ranged from 0.137 to 3.288 μg·L−1, and the M (P25, P75) was 0.808 (0.482, 1.704) μg·L−1. The DCAcAm concentration was the highest (2.448 μg·L−1), followed by BCAcAm, while TCAcAm and DBCAcAm were not detected. The M (P25, P75) of the total concentration of THMs in the drinking water from the Taihu Lake was 33.353 (26.649, 36.217) μg·L−1, that of the Yangtze River was 27.448 (24.312, 31.393) μg·L−1, and both were higher than the level of the reservoir [16.359 (2.305, 21.553) μg·L−1] (P<0.05), while the M (P25, P75) of the total concentration of HAcAms in the drinking water from the Taihu Lake was 0.616 (0.363, 0.718) μg·L−1, which was lower than those of the Yangtze River [0.967 (0.355, 2.283) μg·L−1] and the reservoir [1.071 (0.686, 1.828) μg·L−1] (P<0.05). There were no statistically significant differences in the total concentrations of THMs and HAcAms between wet season and dry season, or between different disinfection methods (P>0.05). The M (P25, P75) concentrations of THMs and HAcAms in drinking water after advanced treatment process involving ozone, activated carbon, and membrane were 20.565 (3.316, 27.185) μg·L−1 and 0.623 (0.452, 1.286) μg·L−1 respectively, and were lower than the corresponding values after conventional treatment process, 28.740 (23.431, 35.085) μg·L−1 and 0.934 (0.490, 2.116) μg·L−1 respectively (P<0.05). Conclusion The concentrations of THMs and HAcAms in drinking water in Wuxi are generally at a low level. The levels of controlled THMs meet the requirements of national standards, and the levels of uncontrolled HAcAms as new DBPs are up to μg·L−1. The concentrations of the two kinds of DBPs in drinking water vary by water sources. The concentrations of THMs and HAcAms produced by the advanced treatment process are lower than that by the conventional treatment process.
4.The effect of maternal vitamin D deficiency on placental steroid hormone synthesis during pregnancy
Lin Fu ; Yuanhua Chen ; Dexiang Xu
Acta Universitatis Medicinalis Anhui 2022;57(4):599-604
Objective:
To explore the mechanism of maternal vitamin D deficiency(VDD)-induced fetal intrauterine growth restriction( IUGR) during pregnancy.
Methods:
Four weeks female CD-1 mice were divided into two groups: control mice( CTRL) and vitamin D deficiency mice( VDD). VDD mice were fed with diet with low concentration of vitamin D and mated with normal male mice. Several pregnant mice were killed on gestational sixteenth day,and the placentas,serum of maternal and fetal mice were collected. Proteins of placental key enzymes of steroid hormone synthesis and signaling pathways of oxidative stress were measured using Western blot. Serum estrogen,progesterone and 25-( OH)-D were measured via radioimmunoassay. The remaining pregnant mice were killed. Placentas and fetus were weighted,fetus height and placental diameter were calculated. The pregnancy outcomes were observed.
Results:
Diet with low concentration of vitamin D were used and VDD mice model were established after five weeks. Maternal VDD during pregnancy decreased the weight of placenta and fetus,reduced the height of fetus and the diameter of placentas. The expression of placental aromatase cytochrome p450( CYP19) and the level of estrogen were increased in VDD pregnant mice. Besides,maternal VDD during pregnancy increased the expressions of placental heme oxygenase 1( HO-1),NADPH oxidase 4( NOX-4) and 3-nitrotyrosine( 3-NT).VDD promoted placental nuclear factor E2-related factor 2( Nrf-2) nuclear translocation.
Conclusion
Maternal vitamin D deficiency during pregnancy causes IUGR partially through evoking placental oxidative stress and promoting estrogen synthesis.
5.Relationship of quality and duration of sleep with hypertension among adults in Guangzhou
Shiyun LUO ; Yongxin YE ; Minying SUN ; Jichuan SHEN ; Nixuan CHEN ; Sheng JIA ; Xueyan LI ; Yuanhua LIU ; Wei ZHU ; Min XIA
Chinese Journal of Preventive Medicine 2021;55(7):853-859
Objective:To explore the relationship of sleep quality and sleep duration with hypertension among adults aged 30-79 years old in Guangzhou.Methods:According to multi-stage stratified cluster sampling, 12 747 residents aged 30-79 years old were sampled and surveyed in Guangzhou from January 2018 to March 2019. Data on general demographic characteristics, sleep quality, sleep duration and hypertension were collected through questionnaire survey, the Pittsburgh sleep quality index (PSQI) and physical examination. Multivariate logistic regression models were used to analyze the putative association between sleep quality, sleep duration and hypertension. Restrictive cubic spline curve was used to draw the dose-response relationship curve between sleep quality, sleep time and hypertension.Results:The mean age of the subjects was (52.68±12.17) years, the prevalence of hypertension was 36.6% (4 664/12 747), the average score of PSQI was (4.70±2.88), and the average sleep time was (7.00±1.32) hours. The prevalence of hypertension was positively associated with the PSQI score. Compared to the subjects with a score less than 3, OR (95% CI) of hypertension with a PSQI score of 3-5, 5-8, ≥9 were 1.14 (1.02-1.27), 1.17 (1.03-1.34), 1.41 (1.21-1.64), respectively. The relationship between sleep duration and hypertension appeared U-shaped. Compared with 6 to 8 hours sleep duration, both sleep duration<6 hours with OR(95% CI) of 1.27(1.12-1.43) or >8 hours with OR(95% CI) of 1.20(1.05-1.38) was associated with hypertension. Conclusion:Both poor sleep quality, longer or shorter sleep duration were responsible for increased risk of cognitive impairment in older Chinese.
6.Relationship of quality and duration of sleep with hypertension among adults in Guangzhou
Shiyun LUO ; Yongxin YE ; Minying SUN ; Jichuan SHEN ; Nixuan CHEN ; Sheng JIA ; Xueyan LI ; Yuanhua LIU ; Wei ZHU ; Min XIA
Chinese Journal of Preventive Medicine 2021;55(7):853-859
Objective:To explore the relationship of sleep quality and sleep duration with hypertension among adults aged 30-79 years old in Guangzhou.Methods:According to multi-stage stratified cluster sampling, 12 747 residents aged 30-79 years old were sampled and surveyed in Guangzhou from January 2018 to March 2019. Data on general demographic characteristics, sleep quality, sleep duration and hypertension were collected through questionnaire survey, the Pittsburgh sleep quality index (PSQI) and physical examination. Multivariate logistic regression models were used to analyze the putative association between sleep quality, sleep duration and hypertension. Restrictive cubic spline curve was used to draw the dose-response relationship curve between sleep quality, sleep time and hypertension.Results:The mean age of the subjects was (52.68±12.17) years, the prevalence of hypertension was 36.6% (4 664/12 747), the average score of PSQI was (4.70±2.88), and the average sleep time was (7.00±1.32) hours. The prevalence of hypertension was positively associated with the PSQI score. Compared to the subjects with a score less than 3, OR (95% CI) of hypertension with a PSQI score of 3-5, 5-8, ≥9 were 1.14 (1.02-1.27), 1.17 (1.03-1.34), 1.41 (1.21-1.64), respectively. The relationship between sleep duration and hypertension appeared U-shaped. Compared with 6 to 8 hours sleep duration, both sleep duration<6 hours with OR(95% CI) of 1.27(1.12-1.43) or >8 hours with OR(95% CI) of 1.20(1.05-1.38) was associated with hypertension. Conclusion:Both poor sleep quality, longer or shorter sleep duration were responsible for increased risk of cognitive impairment in older Chinese.
7.Study on the Quick Daily Check for Medical Electron LINAC.
Yuanhua CHEN ; Juan LI ; Weigang HU ; Xu HAN ; Haiying ZHANG ; Zhiqiang WU
Chinese Journal of Medical Instrumentation 2019;43(2):143-145
This study presents an electronic portal imaging devices (EPIDs) based on daily check tool for Linac that is usable for different cancer centers.Several images of open rectangle fields were acquired with EPID and the key items of daily Linac check were derived from the obtained images using an in-house developed automatic analysis software.The experiment results showed that each parameter calculated by this tool is as reliable as the corresponding result measured by the commercial quality assurance devices and its measuring efficiency is much higher.
Electronics, Medical
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Electrons
;
Particle Accelerators
;
Phantoms, Imaging
;
Radiometry
;
Software
8.Small incision through the tragus for the treatment of sagittal condylar fracture
LIU Yanjie ; LI Gang ; WANG Junlin ; SONG Yong ; CHEN Yucong ; QIN Yuanhua
Journal of Prevention and Treatment for Stomatological Diseases 2018;26(7):460-463
Objective:
The purpose of this study was to a new operative approach for sagittal condylar fractures via a preauricular small incision-based technique and to examine the effectiveness of this approach.
Methods:
Fifteen patients (19 sides) with sagittal condylar fractures were included in the study. The incision length was approximately 4 cm through the tragus, exposing the superficial temporal vessels, which was then pulled forward. Next, the deep temporal superficial fascia was cut, and the surface of the zygomatic arch and the articular capsule of the temporomandibular joint were exposed. Joint capsule incision was performed, with mandibular condylar fracture fixation under direct vision. We followed up with the patients postoperatively for 6 months with clinical and radiographic examinations.
Results :
All patients had 1 week postoperation before being discharged, during which 2 cases of mild facial paralysis (with lateral temporal level Ⅱ facial paralysis, with lateral temporal branch level Ⅲ facial paralysis and level Ⅱ zygomatic branch of facial nerve paralysis after treatment) were observed, after given nerve nutrition agents, 2 cases returned to normal within 3 months. No patient exhibited a postoperative delayed fistula infection or other serious complications. Intraoperative occlusion relationships recovered well, and postoperative CTs suggested that the fracture ends and condyles were in good condition. The occlusion relationship was normal for 3 months after surgery, with a degree of opening greater than 30 mm, no play in the joints and no oblique openings being observed, and reexamination 6 months after the surgery revealed no obvious scars.
Conclusion
This surgical method involves a small incision and clear anatomic structures and avoids damage to the facial nerve. This method provides better surgical vision for treatment of sagittal condylar fractures, is safe and convenient, and deserves clinical recommendation.
9.Clinical Observation of Kangfuxin Liquid in the Treatment of Radioactive Dermatitis of Nasopharyngeal Carcinoma Patients
Zhiping FENG ; Yuanhua SONG ; Zhiyong DENG ; Yaoxiong XIA ; Chao LIU ; Fukun CHEN ; Chuanzhou YANG
China Pharmacy 2018;29(10):1392-1395
OBJECTIVE:To observe clinical efficacy and safety of Kangfuxin liquid in the treatment of radioactive dermatitis of nasopharyngeal carcinoma patients. METHODS:A total of 73 nasopharyngeal carcinoma patients with radioactive dermatitis in the Third Affiliated Hospital of Kunming Medical University during Feb. 2016 to Feb. 2017 were divided into control group (36 cases) and observation group (37 cases) according to random number table. Both groups received routine treatment as Methylrosanilinium chloride solution,Compound cod liver oil and zinc oxide ointment for external use. Observation group was additionally given gauze soaked with Kangfuxin liquid on affected area,3 times a day on the basis of routine treatment. Both groups were treated for 4 weeks. Clinical efficacies of 2 groups were compared,and RTOG classification and dermatitis area were compared before and after treatment. The occurrence of ADR was recorded. RESULTS:There was statistical significance in the total response rate between observation group (94.6%) and control group (97.2%)(P<0.05). Before and after treatment,there were no RTOG Ⅳ patients in 2 groups. Before treatment,there was no statistical significance in the proportion of RTOG 0-Ⅲpatients or dermatitis area between 2 groups (P>0.05). After treatment,RTOG classification of observation group was improved significantly, the proportion of grade 0 patients was significantly higher than control group, that of gradeⅠpatients was significantly lower than control group,with statistical significance(P<0.05). There was no statistical significance in the proportion of RTOG 0-Ⅲpatients of control group before and after treatment(P>0.05). Two,four weeks after treatment,dermatitis areas of 2 groups were decreased significantly;dermatitis area 4 weeks after treatment was significantly smaller than 2 weeks after treatment;observation group was significantly smaller than control group,with statistical significance(P<0.05). No obvious ADR was found in 2 groups during treatment. CONCLUSIONS:Additional use of Kangfuxin liquid in the treatment of radioactive dermatitis of nasopharyngeal carcinoma patients can effectively relieve symptoms and reduce dermatitis area with good safety.
10.VEGF165 transfected endothelial progenitor cells mediated by lentivirus alleviated ALI in rats
Zhaohui HE ; Huiwei HE ; Yuanhua LU ; Zhi CHEN ; Fanghua XU ; Rongsheng WANG ; Chunli YANG
Chinese Critical Care Medicine 2017;29(11):1015-1020
Objective To investigate the protective effects of vascular endothelial growth factor-165 (VEGF165) transfected the endothelial progenitor cells (EPCs) mediated by lentivirus on acute lung injury (ALI) in rats. Methods The mononuclear cells from the male Sprague-Dawley (SD) rats were isolated and cultured to get the EPCs for study. The lentivirus vector carrying the human VEGF165 gene was constructed. According to the random number table method, 90 male SD rats were divided into ALI model group, phosphate buffer solution (PBS) group, EPCs treatment group, none transfected EPCs treatment group and VEGF165 transfected EPCs treatment group, and the rats in each group were subdivided into 4, 12 and 48 hours subgroups, with 6 rats in each subgroup. The rat model of ALI was reproduced by intravenous injection of oleic acid (0.15 μL/g). Then each treatment group was given PBS, EPCs, none transfected EPCs and VEGF165 transfected EPCs respectively with the same volume of 0.2 mL. For the groups with cells, about 1×106cells were contained. Abdominal aortic blood and lung tissue were harvested at 4, 12 and 48 hours. Arterial blood gas analysis was performed. The lung wet/dry weight ratio (W/D) was calculated. The expressions of induced nitric oxide synthase (iNOS), endothelin-1 (ET-1) and VEGF165 were determined by enzyme-linked immunosorbent assay (ELISA). After dyed with hematoxylin-eosin (HE), the lung tissue pathology was observed and the lung injury score was performed. Results Compared with the ALI model group, the arterial partial pressure of oxygen (PaO2) in EPCs, none transfected EPCs and VEGF165 transfected EPCs treatment groups was significantly increased from 4 hours, and lung W/D, expressions of iNOS and ET-1 were significantly decreased, and VEGF165 expression was significantly increased. Compared with the EPCs treatment group, the increase in PaO2, the decrease in lung W/D and expressions of iNOS and ET-1, and the increase in VEGF165 expression in VEGF165 transfected EPCs treatment group were more significant [4 hours: PaO2(mmHg, 1 mmHg = 0.133 kPa) was 82.84±10.69 vs. 72.34±9.36, lung W/D ratio was 4.83±0.23 vs. 5.55±0.37, iNOS (ng/mg) was 8.77±1.10 vs. 14.84±1.34, ET-1 (ng/mg) was 103.41±5.66 vs. 153.08±5.12, VEGF165 (ng/mg) was 130.56±12.16 vs. 83.03±5.95; 12 hours: PaO2(mmHg) was 91.67±6.81 vs. 78.5±8.81, lung W/D ratio was 4.44±0.35 vs. 5.32±0.25, iNOS (ng/mg) was 7.23±0.24 vs. 14.04±1.18, ET-1 (ng/mg) was 91.98±3.52 vs. 125.99±7.55, VEGF165 (ng/mg) was 164.49±5.71 vs. 96.61±6.12]; individual parameters reached valley value or peak value at 48 hours [lung W/D ratio was 4.26±0.30 vs. 4.89±0.15, iNOS (ng/mg) was 5.79±0.85 vs. 12.72±1.10, ET-1 (ng/mg) was 74.53±7.10 vs. 108.33±5.84, VEGF165 (ng/mg) was 237.43±10.79 vs. 134.24±11.99, all P < 0.05]. Over time, lung tissue injury in each group was gradually increased, and the lung injury score was gradually increased. The lung injury score at 48 hours in the EPCs, none transfected EPCs and VEGF165 transfected EPCs treatment groups were lower than that in the ALI model group. Compared with the EPCs treatment group, the VEGF165 transfected EPCs treatment group had a lower score at 48 hours (8.50±1.05 vs. 10.50±1.05, P < 0.05). Conclusion The transplantation of EPCs which were transfected with VEGF165 mediated by lentivirus could obviously improve the oxygen pressure, reduce the lung water seepage, decrease the iNOS and ET-1 expressions in lung tissue, and had obvious protective effects on ALI.


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