1.Radon concentration and related employees’ chromosome aberration and micronuclei in subways in a city of Zhejiang Province
Zhiqiang XUAN ; Xinyu HUANG ; Zhongjun LAI ; Jiadi GUO ; Xiaoji HAO ; Shunfei YU ; Yiyao CAO ; Donghang WANG ; Duo ZHANG ; Jihua NIE
Journal of Environmental and Occupational Medicine 2024;41(9):1025-1031
		                        		
		                        			
		                        			Background Excessive radon exposure is considered the second risk factor for lung cancer. Since the opening of the subway in a city of Zhejiang Province, the exposure level of radioactive gas radon in subway stations and its impact on occupational health have become one of the important issues of public concern. Objective To monitor the radon concentration of subways in a city in Zhejiang Province and explore the effect of radon exposure on chromosome aberration and micronuclei in the working population. Methods A total of 55 vehicle control rooms of 55 stations affiliated to two different subway lines in a city were measured for one year; the 110 ticket offices and 55 security checkpoints from the same 55 stations were measured from 16 March to 14 June. The radon concentrations were compared by job types, subway lines, and seasons referring to Measurement methods for determination of radon in environmental air (HJ 1212-2021). Peripheral blood lymphocyte chromosome aberration and micronucleus analyses were conducted in 165 subway workers from monitoring sites for three different job types, then the influencing factors were analyzed. The detection methods were adopted from the standards of Test and assessment of chromosomal aberrations on occupational health examinations for radiation workers (GBZ/T 248-2014) and Standard for the method of micronucleus detection in lymphocytes on occupational health examination for radiation workers and exposure dose estimation (GBZ/T 328-2023). Results The radon concentration range of the target subways in Zhejiang Province was 10-320 Bq·m−3, all lower than the national limit (≤400 Bq·m−3). The differences in radon radioactivity levels among different lines, job types, and time segments were statistically significant (P<0.05). The rates of chromosomal aberration and micronucleus formation among the 165 subjects were 0.224% and 0.024%, respectively. There were significant differences in the rates of chromosome aberration and micronuclei among different jobs (vehicle control room, ticket office, security checkpoint) (P<0.05), but the abnormal rates were lower than the limits of the corresponding national standard. No significant correlation was found between jobs and chromosomal aberrations or micronuclei (P>0.05). Chromosome aberration and micronuclei varied by age, subway station seniority, and smoking (P<0.05). No effect of the above factors on chromosome aberration and micronuclei was observed by logistic regression (P>0.05). Conclusion The radon concentration in the target subway system is at a normal level. The rates of chromosomal aberration and micronucleus formation vary by jobs, but both are lower than the corresponding national limits. Therefore, radon exposure has not yet caused outstanding health impact on the subway workers.
		                        		
		                        		
		                        		
		                        	
2.Analysis of the current situation of occupational health among radiation workers in some areas of Zhejiang Province, China
Xiaoji HAO ; Zhiqiang XUAN ; Jiadi GUO ; Zhongjun LAI ; Donghang WANG ; Shunfei YU
Chinese Journal of Radiological Health 2024;33(5):549-554
		                        		
		                        			
		                        			Objective To investigate the current situation of occupational health of radiation workers in Zhejiang Province, China, and to provide a theoretical basis for the improvement of standards and management systems for occupational health examinations of radiation workers. Methods Data of occupational health examination cases were collected from occupational health examination institutions, including basic information, type of physical examination (pre-job, on-job, off-job), and health examination results. The differences in occupational health examination results among different groups were analyzed, and the factors influencing occupational health of radiation workers were analyzed using a logistic regression model. Results A total of 
		                        		
		                        	
3.Analysis of the current situation of occupational health among radiation workers in some areas of Zhejiang Province, China
Xiaoji HAO ; Zhiqiang XUAN ; Jiadi GUO ; Zhongjun LAI ; Donghang WANG ; Shunfei YU
Chinese Journal of Radiological Health 2024;33(5):549-554
		                        		
		                        			
		                        			Objective To investigate the current situation of occupational health of radiation workers in Zhejiang Province, China, and to provide a theoretical basis for the improvement of standards and management systems for occupational health examinations of radiation workers. Methods Data of occupational health examination cases were collected from occupational health examination institutions, including basic information, type of physical examination (pre-job, on-job, off-job), and health examination results. The differences in occupational health examination results among different groups were analyzed, and the factors influencing occupational health of radiation workers were analyzed using a logistic regression model. Results A total of 
		                        		
		                        	
4.To explore the effect of A2AR on endotoxin-induced inflammatory injury of Caco-2 intestinal epithelial cells from the perspective of autophagy
Yu CHEN ; Xiaoji WANG ; He LI
Acta Universitatis Medicinalis Anhui 2024;59(9):1636-1642
		                        		
		                        			
		                        			Objective To explore the effect and mechanism of adenosine A2A receptor(A2AR)on lipopolysac-charide(LPS)-induced inflammatory injury of Caco-2 intestinal epithelial cells.Methods Caco-2 cells were di-vided into control group,LPS group(treated with 10 μg/ml LPS for 12 h),A2AR agonist(CGS21680)group(pretreated with 50 nmol/L CGS21680 for 10 min),CGS21680+LPS group(pretreated with 50 nmol/L CGS21680 for 10 min,treated with 10 μg/ml LPS for 12 h),cell viability was determined using CCK-8 assay,the secretion levels of tumor necrosis factor-α(TNF-α),interleukin-1 β(IL-1 β)and interleukin-6(IL-6)in cell su-pernatant of each group were determined using ELISA.mRNA expression levels of TNF-α,IL-1 β and IL-6 in cells of each group were detected by real-time fluorescence quantitative PCR,the protein expression levels of microtubule associated light chain protein 3(LC3)-Ⅱ/LC3-Ⅰ and autophagy associated protein(Beclin1)in cells of each group were analyzed using Western blot analysis.Caco-2 cells were then divided into control group,LPS group(pretreated with 50 nmol/L CGS21680 for 10 min),CGS21680+LPS group(pretreated with 50 nmol/L CGS21680 for 10 min,treated with 10 μg/ml LPS for 12 h),CGS21680+LPS+Rapa group(pretreated with 50 nmol/L CGS21680 for 10 min,treated with 10 μg/ml LPS and 5 μmol/L Rapa for 12 h),cell viability was deter-mined using CCK-8 assay,the secretion levels of TNF-α,IL-1 β and IL-6 in cell supernatant of each group were determined using ELISA.Results Compared with the control group,the viability of Caco-2 cells in LPS group sig-nificantly decreased(P<0.05),the levels of TNF-α,IL-1 β and IL-6 in supernatant significantly increased(P<0.05),the mRNA relative expressions of TNF-α,IL-1β,IL-6 in cells significantly increased(P<0.05),the LC3-Ⅱ/LC3-Ⅰ ratio and the relative expression of Beclin1 protein were significantly up-regulated(P<0.05).Compared with LPS group,the viability of Caco-2 cells in CGS21680+LPS group significantly increased(P<0.05),the levels of TNF-α,IL-1 β and IL-6 in supernatant significantly decreased(P<0.05),the mRNA rela-tive expression levels of TNF-α,IL-1β,IL-6 in cells were significantly down-regulated(P<0.05),and the ratio of LC3-Ⅱ/LC3-Ⅰ and the relative expression of Beclin1 protein were significantly down-regulated(P<0.05).In addition,compared with CGS21680+LPS group,the viability of Caco-2 cells in CGS21680+LPS+Rapa group significantly decreased(P<0.05),and the levels of TNF-α,IL-1 β and IL-6 in the supernatant also significantly increased(P<0.05).Conclusion A2 AR agonist can reduce the inflammatory injury of Caco-2 intestinal epithe-lial cells induced by LPS and improve cell viability,which may be related to its inhibition of autophagy.
		                        		
		                        		
		                        		
		                        	
5. Application value of real-time virtual sonography in diagnosis and treatment of complicated hepatolithiasis
Yan XIAO ; Lei ZHOU ; Wei CHENG ; Shengtao HUANG ; Yaling LIU ; Guoguang LI ; Jianhui YANG ; Xiaoji PAN ; Xiangying WANG
Chinese Journal of Digestive Surgery 2020;19(1):99-105
		                        		
		                        			 Objective:
		                        			To investigate the application value of real-time virtual sonography(RVS)in the diagnosis and treatment of complicated hepatolithiasis.
		                        		
		                        			Methods:
		                        			The retrospective and descriptive study was conducted. The clinical data of 10 patients with complicated hepatolithiasis who were admitted to Hunan Provincial People′s Hospital between October 2017 and March 2018 were collected. There were 3 males and 7 females, aged from 40 to 69 years, with an average age of 57 years. Patients received abdominal color Doppler ultrasound examination, magnetic resonance cholangiopancreatography, and upper abdominal spiral computed tomography (CT) thinly scanning + enhanced examination. Data of CT examination were imported into RVS. RVS was used to locate hepatolithiasis, relationship between stones and vessels, anatomy of bile ducts and vessels in hepatic hilus. Surgical methods included RVS-guided hilar cholangiotomy, biliary stricturoplasty, bilateral hepatojejunostomy, hepatic segmentectomy (lobectomy), and hepatolithotomy. Observation indicators: (1) surgical and postoperative situations; (2) typical case analysis; (3) follow-up. Follow-up using outpatient examination was performed to detect residual stones up to June 2019. Measurement data with skewed distribution were represented as 
		                        		
		                        	
6. LncRNA HULC promots HCC growth by downregulating miR-29
Liru ZHU ; Jianli FENG ; Xiaoji LIU ; Junmei WANG
Chinese Journal of Oncology 2019;41(9):659-666
		                        		
		                        			 Objective:
		                        			To explore the effects of lncRNA HULC on hepatocellular carcinoma (HCC) growth by down-regulating miR-29.
		                        		
		                        			Methods:
		                        			The expression levels of HULC and miR-29 in HCC tissues and cells were detected by real-time quantitative PCR (RT-qPCR), and the correlation analysis was performed. After HCC cells were transfected with HULC overexpressed plasmid or siRNA, the expressions of miR-29 and its target gene SETDB1 were determinate by RT-qPCR. According to the bioinformatic prediction of the miR-29 binding site in the HULC sequence, the report gene plasmids were constructed. The HCC cells were co-transfected with miR-29 mimics or miR-29 inhibitor, and the HULC targeted regulation of miR-29 was verified by dual luciferase reporter assay. The effect of miR-29 on the HULC-mediated proliferation in HCC cells was detected by cell count kit 8 (CCK-8) experiment. Expression of tumor proliferation antigen Ki-67 was detected by RT-qPCR.The Hep3B cells were inoculated in mice and miR-29 mimics and miR-29 negative control (NC) further injected into the lesions. The tumor volume was observed, and the expressions of tumor proliferation antigen ki-67 in tumor tissues were detected by immunohistochemical staining.
		                        		
		                        			Results:
		                        			The expression of HULC was significantly up-regulated while the expression of miR-29 was significantly down-regulated in HCC tissues and cells (
		                        		
		                        	
7.LncRNA HULC promots HCC growth by downregulating miR?29
Liru ZHU ; Jianli FENG ; Xiaoji LIU ; Junmei WANG
Chinese Journal of Oncology 2019;41(9):659-666
		                        		
		                        			
		                        			Objective To explore the effects of lncRNA HULC on hepatocellular carcinoma (HCC) growth by down?regulating miR?29. Methods The expression levels of HULC and miR?29 in HCC tissues and cells were detected by real?time quantitative PCR ( RT?qPCR ), and the correlation analysis was performed. After HCC cells were transfected with HULC overexpressed plasmid or siRNA, the expressions of miR?29 and its target gene SETDB1 were determinate by RT?qPCR. According to the bioinformatic prediction of the miR?29 binding site in the HULC sequence, the report gene plasmids were constructed.The HCC cells were co?transfected with miR?29 mimics or miR?29 inhibitor, and the HULC targeted regulation of miR?29 was verified by dual luciferase reporter assay. The effect of miR?29 on the HULC?mediated proliferation in HCC cells was detected by cell count kit 8 ( CCK?8) experiment. Expression of tumor proliferation antigen Ki?67 was detected by RT?qPCR.The Hep3B cells were inoculated in mice and miR?29 mimics and miR?29 negative control (NC) further injected into the lesions. The tumor volume was observed, and the expressions of tumor proliferation antigen ki?67 in tumor tissues were detected by immunohistochemical staining. Results The expression of HULC was significantly up?regulated while the expression of miR?29 was significantly down?regulated in HCC tissues and cells (P<0.01). The level of HULC was negatively correlated with miR?29 in tumor tissues (r=-0.754, P<0.01) and HCC cells ( r=-0.865, P<0.05).The in vitro experiments showed that, compared with the blank control group, the expression of miR?29 in HULC overexpressed Huh7 cells was significantly reduced, while the mRNA level of miR?29 target gene SETDB1 was increased ( P<0.01). The expression of miR?29 was significantly increased in HULC deleted Hep3B cells, while the mRNA expression of SETDB1 was decreased (P<0.01). Double luciferase reporter gene assay showed that miR?29 mimics significantly inhibited the luciferase activity of Hep3B cells transfected with HULC wide type ( psi?HULC?WT) plasmid but had no effect on Hep3B cells transfected with mutant plasmid ( psi?HULC?Mut). However, the miR?29 inhibitor antagonized the inhibitory effect of miR?29 mimics on luciferase activity of psi?HULC?WT (P<0.01).Cell proliferation experiments showed that, compared with the control group, the proliferation ability of miR?29 mimics overexpressed Huh7 cells was significantly reduced.After 24, 48 and 72 hours of treatment, the proliferation rates of Huh7 cells in the HULC overexpressed group were (43.87± 3.82)%, (83.45±7.46)% and (123.34±8.67)%, respectively, significantly higher than (13.45±1.77)%, (23.54±1.37)% and (38.21±2.09)% of control group (P<0.05).After treatment for 48 and 72 hours, the proliferation rates of miR?29 mimics transfected Huh7 cells were ( 57.10 ± 1.94)% and ( 73.76± 3.46)%, respectively, significantly lower than (83.45±7.46)% and ( 123.34±8.67)% of control group ( P<0.05). After treatment for 48 and 72 hours, the proliferation rates of Huh7 cells transfected with miR?29 mimics and miR?29 inhibitor group were (76.45± 3.24)% and ( 89.37± 4.37)%, respectively, significant higher than (57.10%±1.94)% and ( 73.76 ± 3.46)% of the control group ( P<0.05). After 48 h transfection, the expression of Ki?67 in Huh7 transfected with miR?29 mimics was significantly inhibited compared with the control group (P<0.01). However, the expression of Ki?67 mRNA was increased in Huh7 cells transfected with miR?29 inhibitor (P<0.01). The results of in vivo experiments showed that the tumor volumes of the control group, miR?29 mimics group and miR?29 mimics + miR?29 inhibitors group were ( 504.0± 19.6) mm3, (310.0±24.3) mm3 and (483.7±21.2) mm3, respectively. Injection of miR?29 mimics reduced while miR?29 inhibitor promoted tumorigenesis ability of Huh7 in nude mice (P<0.01). The immunohistochemical staining showed that the average optical density values of Ki?67 protein in tumor tissues of the control group, miR?29 mimics group and miR?29 analogue+miR?29 inhibitor group were 0.65±0.08, 0.36±0.07 and 0.56±0.06, respectively. The expression level of Ki?67 protein in miR?29 mimics group was significantly reduced ( P<0.01) while increased in the miR?29 mimics+miR?29 inhibitor group ( P<0.01). Conclusion LncRNA HULC promotes HCC growth by down?regulating miR?29.
		                        		
		                        		
		                        		
		                        	
8.LncRNA HULC promots HCC growth by downregulating miR?29
Liru ZHU ; Jianli FENG ; Xiaoji LIU ; Junmei WANG
Chinese Journal of Oncology 2019;41(9):659-666
		                        		
		                        			
		                        			Objective To explore the effects of lncRNA HULC on hepatocellular carcinoma (HCC) growth by down?regulating miR?29. Methods The expression levels of HULC and miR?29 in HCC tissues and cells were detected by real?time quantitative PCR ( RT?qPCR ), and the correlation analysis was performed. After HCC cells were transfected with HULC overexpressed plasmid or siRNA, the expressions of miR?29 and its target gene SETDB1 were determinate by RT?qPCR. According to the bioinformatic prediction of the miR?29 binding site in the HULC sequence, the report gene plasmids were constructed.The HCC cells were co?transfected with miR?29 mimics or miR?29 inhibitor, and the HULC targeted regulation of miR?29 was verified by dual luciferase reporter assay. The effect of miR?29 on the HULC?mediated proliferation in HCC cells was detected by cell count kit 8 ( CCK?8) experiment. Expression of tumor proliferation antigen Ki?67 was detected by RT?qPCR.The Hep3B cells were inoculated in mice and miR?29 mimics and miR?29 negative control (NC) further injected into the lesions. The tumor volume was observed, and the expressions of tumor proliferation antigen ki?67 in tumor tissues were detected by immunohistochemical staining. Results The expression of HULC was significantly up?regulated while the expression of miR?29 was significantly down?regulated in HCC tissues and cells (P<0.01). The level of HULC was negatively correlated with miR?29 in tumor tissues (r=-0.754, P<0.01) and HCC cells ( r=-0.865, P<0.05).The in vitro experiments showed that, compared with the blank control group, the expression of miR?29 in HULC overexpressed Huh7 cells was significantly reduced, while the mRNA level of miR?29 target gene SETDB1 was increased ( P<0.01). The expression of miR?29 was significantly increased in HULC deleted Hep3B cells, while the mRNA expression of SETDB1 was decreased (P<0.01). Double luciferase reporter gene assay showed that miR?29 mimics significantly inhibited the luciferase activity of Hep3B cells transfected with HULC wide type ( psi?HULC?WT) plasmid but had no effect on Hep3B cells transfected with mutant plasmid ( psi?HULC?Mut). However, the miR?29 inhibitor antagonized the inhibitory effect of miR?29 mimics on luciferase activity of psi?HULC?WT (P<0.01).Cell proliferation experiments showed that, compared with the control group, the proliferation ability of miR?29 mimics overexpressed Huh7 cells was significantly reduced.After 24, 48 and 72 hours of treatment, the proliferation rates of Huh7 cells in the HULC overexpressed group were (43.87± 3.82)%, (83.45±7.46)% and (123.34±8.67)%, respectively, significantly higher than (13.45±1.77)%, (23.54±1.37)% and (38.21±2.09)% of control group (P<0.05).After treatment for 48 and 72 hours, the proliferation rates of miR?29 mimics transfected Huh7 cells were ( 57.10 ± 1.94)% and ( 73.76± 3.46)%, respectively, significantly lower than (83.45±7.46)% and ( 123.34±8.67)% of control group ( P<0.05). After treatment for 48 and 72 hours, the proliferation rates of Huh7 cells transfected with miR?29 mimics and miR?29 inhibitor group were (76.45± 3.24)% and ( 89.37± 4.37)%, respectively, significant higher than (57.10%±1.94)% and ( 73.76 ± 3.46)% of the control group ( P<0.05). After 48 h transfection, the expression of Ki?67 in Huh7 transfected with miR?29 mimics was significantly inhibited compared with the control group (P<0.01). However, the expression of Ki?67 mRNA was increased in Huh7 cells transfected with miR?29 inhibitor (P<0.01). The results of in vivo experiments showed that the tumor volumes of the control group, miR?29 mimics group and miR?29 mimics + miR?29 inhibitors group were ( 504.0± 19.6) mm3, (310.0±24.3) mm3 and (483.7±21.2) mm3, respectively. Injection of miR?29 mimics reduced while miR?29 inhibitor promoted tumorigenesis ability of Huh7 in nude mice (P<0.01). The immunohistochemical staining showed that the average optical density values of Ki?67 protein in tumor tissues of the control group, miR?29 mimics group and miR?29 analogue+miR?29 inhibitor group were 0.65±0.08, 0.36±0.07 and 0.56±0.06, respectively. The expression level of Ki?67 protein in miR?29 mimics group was significantly reduced ( P<0.01) while increased in the miR?29 mimics+miR?29 inhibitor group ( P<0.01). Conclusion LncRNA HULC promotes HCC growth by down?regulating miR?29.
		                        		
		                        		
		                        		
		                        	
9.Morphological assessment of sulfur mustard-induced acute lung injury in rats through different routes.
Shuangshuang ZHU ; Xiaoji ZHU ; Jian ZHAO ; Yuxu ZHONG ; Hua YANG ; Tao WANG ; Peng JI ; Ling-Yan JU ; Meihong WANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2015;33(9):685-688
OBJECTIVETo establish an animal model of sulfur mustard (SM)-induced acute lung injury in rats through different routes and compare the morphological changes in lung tissue and cells.
METHODSOne hundred and thirty-six male rats were selected and randomly divided into 5 groups, namely peritoneal cavity SM group (n=32), trachea SM group (n=32), peritoneal cavity propylene glycol group (n=32), trachea propylene glycol group (n=32), and normal control group (n=8). The rats in peritoneal cavity SM group were injected intraperitoneally with diluted SM (0.1 ml, 8 mg/kg), and the rats in trachea SM group were injected intratracheally with diluted SM (0.1 ml, 2 mg/kg). Once the rats were sacrificed at 6, 24, 48, and 72 h after SM treatment, morphological changes in lung tissue and cells were observed by light and electron microscopy.
RESULTSIn the peritoneal cavity SM group, the epithelial cells of bronchioles maintained intact with increased exudate and bleeding in alveolar cavity and large areas of pulmonary consolidation under the light microscope. In the tracheal SM group, focal ulcer formed in the epithelial cells of bronchioles with increased exudate and bleeding in alveolar cavity, partial pulmonary consolidation, and compensatory emphysema in peripheral alveolar space under the light microscope. The alveolar interval areas were widened obviously in both groups in a time-dependent manner. Under the electron microscope, we observed local loss of cellular membrane in type I alveolar epithelium, broken or lost microvilli in cells of typeⅡalveolar epithelium and fuzzy mitochondrial crista as well as the appearance of ribosome detached from rough endoplasmic reticulum in both two groups. Compared with those in the trachea SM group and the control group, the ratio of the alveolar septum average area to the visual field area in the peritoneal cavity SM group at 6, 24, 48, and 72 h was significantly higher (P<0.05).
CONCLUSIONThe lung tissue injury through the intraperitoneal route is more severe than that through the tracheal route, while focal ulceration of bronchioles epithelial cells appears in the case of tracheal route. The degree of injury increases over time in both groups, and the cellular damage is approximately the same in both groups.
Acute Lung Injury ; chemically induced ; pathology ; Animals ; Disease Models, Animal ; Lung ; pathology ; Male ; Mustard Gas ; toxicity ; Peritoneum ; Pulmonary Alveoli ; pathology ; ultrastructure ; Rats ; Trachea
10.Correlation between liver damage degree and clinical manifestations in patients with pulmonary tuberculosis after chemotherapy
Ying SUN ; Runpu LI ; Xiaoji WANG
Journal of Clinical Hepatology 2015;31(7):1110-1113
		                        		
		                        			
		                        			 ObjectiveTo explore the correlation between the degree of liver damage and clinical manifestations in patients with pulmonary tuberculosis after chemotherapy. MethodsThis study included 3620 new smear-positive pulmonary tuberculosis patients treated with first-line anti-tuberculosis drug in the Second Central Hospital of Baoding from January 2008 to January 2014, and the follow-up study was carried out to observe medication use and side effects of anti-tuberculosis drug treatment. Comparison of categorical data was made by chisquare test. ResultsA total of 1225 patients (33.8%) exhibited clinical manifestations related to liver injury. The most common clinical manifestation was nausea and vomiting (72.9%), followed by fatigue (37.8%), rash (31.5%), abdominal distension and diarrhea (281%), fever (14.2%), anorexia (3.8%), and other manifestations (2.0%). The nausea and vomiting usually first appeared and were followed by abdominal distension and diarrhea. Of all patients, 243 cases (6.7%) suffered from liver damage and 109 cases (3.0%) had moderate to severe liver damage. Of the patients with clinical manifestations, 171 cases (14.0%) had liver damage and 74 cases(60%) suffered from moderate to severe liver damage. Compared with the patients without clinical manifestations, the relative risks of liver damage and moderate to severe liver damage were 4.643 [95% confidence interval (CI)=3.035-4.856] and 4.134 (95% CI=2.817-4.351), respectively, in the patients with clinical manifestations. The patients with fatigue, nausea and vomiting, rash, abdominal distension and diarrhea, anorexia, and other manifestations had higher risk of liver damage and moderate to severe liver damage than those without clinical manifestations (P<0.05) and the patients with anorexia showed the highest risk. ConclusionOne third of patients with pulmonary tuberculosis have liver injury-related clinical manifestations after chemotherapy. Patients with fatigue, nausea and vomiting, rash, abdominal distension and diarrhea, anorexia, and other manifestations are more susceptible to liver damage and moderate to severe liver damage. 
		                        		
		                        		
		                        		
		                        	
            

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