1.Clinical application analysis of transumbilical single-port laparoscopic hepatectomy
Dongdong DAI ; Tengfei QU ; Siyu MOU ; Tinglong ZHANG ; Lin XU ; Jinzhong PANG ; Puwen LIU ; Jinchuan ZHAO ; Zusen WANG
Chinese Journal of Hepatobiliary Surgery 2024;30(7):494-498
		                        		
		                        			
		                        			Objective:To explore the efficacy, safety, and feasibility of single port laparoscopic liver resection via umbilical cord, and summarize its surgical experience.Method:A retrospective analysis was conducted on 39 patients who underwent liver resection surgery at the Department of Hepatobiliary and Pancreatic Surgery, the Affiliated Hospital of Qingdao University from February 2022 to September 2023. There were 19 patients in the transumbilical single-port laparoscopic group, including 5 males and 14 females, aged (49.6±2.5) years. There were 20 patients in the multi-port laparoscopic group, including 7 males and 13 females, aged (49.9±3.1) years. The intraoperative blood loss, operation time, intestinal recovery time, postoperative hospital stay and postoperative complications were compared between the single-port group and multi-port group.Results:All 39 patients successfully completed the surgery without any additional foramen or conversion to open surgery. The operation time of the single hole group (166.3±59.0) min was longer than that of the multi-port group (123.2±48.0) min, and the difference was statistically significant ( t=2.50, P=0.020). There were no statistically significant differences in intraoperative blood loss, intestinal recovery time, postoperative hospital stay, and postoperative complications between these two groups (all P>0.05). All patients had no postoperative complications such as bleeding, infection, or bile leakage. Follow up for 3~21 months showed no recurrence of primary diseases such as hepatic hemangioma, hepatic adenoma, and intrahepatic bile duct stones. The aesthetic effect of the umbilical incision in the single orifice group was significant, and patient satisfaction was 100%. Conclusion:Umbilical single-port laparoscopic liver resection surgery is safe and feasible, with significant minimally invasive and aesthetic effects.
		                        		
		                        		
		                        		
		                        	
2.Mental health disparities in people living with human immunodeficiency virus: A cross-sectional study on physician-patient concordance and treatment regimens
Jinchuan SHI ; Zhongdong ZHANG ; Junyan ZHANG ; Yishu ZHANG ; Jiating QIU ; Fang LIU ; Daoyuan SONG ; Yanfang MA ; Lianmei ZHONG ; Hongxing WANG ; Xiaolei LIU
Chinese Medical Journal 2024;137(18):2223-2232
		                        		
		                        			
		                        			Background::Human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) can profoundly affect the mental health of the people living with HIV (PLWH), with higher rates of anxiety, depression, and sleep disturbances. The disparities in neuropsychological problems evaluated by physicians and self-assessed by patients are still unknown.Methods::A total of 5000 PLWH and 500 physicians from 167 hospitals were enrolled in this cross-sectional study from September 2022 to February 2023. 4-Item Patient Health Questionnaire (PHQ-4) was used for the evaluation of depressive issues and anxiety issues by PLWH. Each physician assessed 10 PLWH under their care for the presence of depressive or anxiety issues. The primary outcomes of this study are the concordance rates on the depressive issues and anxiety issues evaluation between physicians and PLWH. The Cohen’s kappa test was used to assess the agreement between physicians and PLWH.Results::The concordance rate for the evaluation of depressive issues is 73.84% (95% confidence interval [CI]: 72.60-75.04%), and it is significantly different from the expected rate of 80% ( P <0.001). Similarly, the concordance rate for the evaluation of anxiety issues is 71.74% (95% CI: 70.47-72.97%), which is significantly different from the expected rate of 80% as per the null hypothesis ( P <0.001). The overestimation rate by physicians on depressive issues is 12.20% (95% CI: 11.32-13.14%), and for anxiety issues is 12.76% (95% CI: 11.86-13.71%). The mismatch rate for depressive issues is 26.16% (95% CI: 24.96-27.40%), and for anxiety issues is 28.26% (95% CI: 27.02-29.53%). The underestimation rate by physicians on depressive issues is 13.96% (95% CI: 13.03-14.95%), and for anxiety issues is 15.50% (95% CI: 14.52-16.53%). For the treatment regiments, PLWH sustained on innovative treatment regimen (IR) related to a lower prevalence of depressive issues (odds ratio [OR] = 0.71, 95% CI: 0.59-0.87, P = 0.003) and a lower prevalence of anxiety issues (OR = 0.63, 95% CI: 0.52-0.76, P <0.001). PLWH switch from conventional treatment regimen (CR) to IR also related to a lower prevalence of depressive issues (OR = 0.79, 95% CI: 0.64-0.98) and a lower prevalence of anxiety issues (OR = 0.81, 95% CI: 0.67-0.99). Conclusion::Nearly one in three PLWH had their condition misjudged by their physicians. The findings underscore the need for improved communication and standardized assessment protocols in the care of PLWH, especially during the acute phase of HIV infection.
		                        		
		                        		
		                        		
		                        	
3.Effect of Dahuang Xiezhuo Prescription on Inflammation and SOCS3/TLR4 Pathway in Rats with Chronic Renal Failure
Xin XIN ; Zhenhua WU ; Miao TAN ; Fei GAO ; Fengwen YANG ; Suzhi CHEN ; Qian ZHANG ; Jinchuan TAN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(7):95-104
		                        		
		                        			
		                        			ObjectiveTo observe the intervention effect of Dahuang Xiezhuo prescription (DHXZ) on inflammation and suppressor of cytokine signaling 3 (SOCS3)/Toll-like receptor 4 (TLR4) pathway in rats with chronic renal failure (CRF), and to explore its molecular mechanism in alleviating renal inflammatory response. MethodThe 90 male SD rats, 15 were randomly selected as sham group, and the remaining 75 were used as modeling group to replicate CRF rat model by 5/6 nephrectomy. After successful modeling, the rats were randomly divided into model group, DHXZ low-, medium-, high-dose groups (6.825, 13.65, 27.3 g·kg-1) and Niaoduqing Granules group (2.6 g·kg-1). The drug intervention groups received corresponding drugs by gavage for 8 consecutive weeks. After administration, hematoxylin-eosin (HE) staining and Masson staining were used to observe the morphological changes of rat renal tissue, and blood creatinine (SCr), blood urea nitrogen (BUN) and blood uric acid (UA) were tested. Enzyme-linked immunosorbent assay (ELISA) was performed to detect the serum contents of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and C-reactive protein (CRP). The mRNA expressions of SOCS3 and TLR4 in renal tissue were detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), and the protein expressions of SOCS3, TLR4, nuclear transcription factor (NF-κB) and myeloid differentiation factor (MyD88) were detected by Western blot. Immunohistochemistry was used to determine the protein expressions of NF-κB, MyD88, NOD-like receptor protein 3 (NLRP3) and melanoma deficiency factor 2 (AIM2). ResultCompared with the sham group, the model group had a significant inflammatory response in renal tissue, and an increase in blood SCr, BUN, UTP, IL-6, TNF-α and CRP (P<0.05). The protein and mRNA expressions of SOCS3 in renal tissue of rats in the model group were lower while the protein expressions of TLR4, NF-κB, MyD88, NLRP3 and AIM2 and the mRNA expression of TLR4 were higher than those in the sham group (P<0.05). Compared with the model group, DHXZ and Niaoduqing granules groups presented markedly reduced inflammatory response in renal tissue and decreased blood SCr, BUN, UTP, IL-6, TNF-α and CRP (P<0.05). Additionally, DHXZ and Niaoduqing granules up-regulated the protein and mRNA expressions of SOCS3 in renal tissue while down-regulated the protein expressions of TLR4, NF-κB, MyD88, NLRP3 and AIM2 and the mRNA expression of TLR4 (P<0.05). ConclusionDHXZ can reduce the release and expression of inflammatory factors, inhibit the inflammatory response and improve renal function, and the mechanism may be related to the regulation of SOCS3/TLR4 signaling pathway. 
		                        		
		                        		
		                        		
		                        	
4.Mechanism of Salvianolate in Inducing Autophagy in Podocytes of Rats with Membranous Nephropathy via AMPK/SIRT1/PGC-1α Signaling Pathway
Yao ZHANG ; Fei GAO ; Miao TAN ; Fengwen YANG ; Suzhi CHEN ; Meifang REN ; Guodong YUAN ; Jinchuan TAN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(8):203-213
		                        		
		                        			
		                        			ObjectiveTo observe the effect of salvianolate on the protein expressions of adenosine monophosphate (AMP)-activated protein kinase (AMPK), silent information regulator 1 (SIRT1) and peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α), autophagy and apoptosis in kidney tissue of rats with membranous nephropathy (MN), and to explore its possible molecular mechanism against MN. MethodEighty male SD rats were randomly divided into normal group, model group, benazepril hydrochloride group (10 mg·kg-1), and salvianolate low-, medium-, and high-dose groups (16.7, 33.3 and 66.7 mg·kg-1). The rats were modeled by injection of cationized bovine serum albumin (C-BSA) into the tail vein. After successful modeling, rats in the administration groups were given corresponding doses of drugs for 4 consecutive weeks, and then 24-hour urine, serum and kidney tissue were collected for the detection of 24-hour urinary protein (UTP), blood urea nitrogen (BUN), serum creatinine (SCr), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), C reactive protein (CRP), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), and malondialdehyde (MDA). The pathological changes of kidneys were observed by light microscope, electron microscope and immunofluorescence. Western blot was used to detect the protein expressions of phospho-AMPK (p-AMPK), AMPK, phospho-SIRT1 (p-SIRT1), SIRT1 and PGC-1α in rat kidney tissue. The protein expressions of autophagy-specific gene (Beclin-1), microtubule-associated protein 1 light chain 3 (LC3) Ⅱ, ubiquitin-binding protein (p62), B cell lymphoma (Bcl-2), Bcl-2-associated X (Bax), and cysteine aspartic protease-7 (Caspase-7) in rat kidney tissue were determined by immunohistochemistry (IHC). ResultCompared with the conditions in the normal group, the levels of UTP, IL-6, TNF-α, CRP and MDA in the model group were increased (P<0.05) while the levels of SOD and GSH-Px were decreased (P<0.05), and there was no difference in BUN and SCr. Compared with the model group, the administration groups had lowered UTP, IL-6, TNF-α, CRP and MDA (P<0.05) while elevated SOD and GSH-Px (P<0.05). It could be seen from hematoxylin and eosin (HE) staining, Masson staining, immunofluorescence and electron microscopy that the pathological damage of rat kidney tissue in the model group was significant, but after treatment with benazepril hydrochloride and salvianolate, the pathological damage of kidney cells was gradually improved. The expressions of p-AMPK/AMPK, p-SIRT1/SIRT1, PGC-1α, Bcl-2, Beclin-1 and LC3Ⅱ in rat kidney in the model group were lower than those in the normal group (P<0.05) while the expressions of Bax, Caspase-7 and p62 were higher (P<0.05). Compared with the model group, benazepril hydrochloride group and salvianolate groups had an up-regulation in the expressions of p-AMPK/AMPK, p-SIRT1/SIRT1, PGC-1α, Bcl-2, Beclin-1 and LC3Ⅱ in the kidney (P<0.05) while a down-regulation in the expressions of Bax, Caspase-7 and p62 (P<0.05). ConclusionThe protective effect of salvianolate on the kidneys of MN rats may be related to the activation of AMPK/SIRT1/PGC-1α signaling pathway, the up-regulation of autophagy and the reduction of apoptosis. 
		                        		
		                        		
		                        		
		                        	
5.Effect of Dahuang Xiezhuo Prescription on Renal Tubular Function in Patients with IgA Nephropathy Complicated with Hyperuricemia
Qian ZHANG ; Fengwen YANG ; Meng SHI ; Jinchang LIU ; Gang WANG ; Meifang REN ; Meijiao ZHAO ; Jingyu MAO ; Jinchuan TAN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(12):135-141
		                        		
		                        			
		                        			ObjectiveTo observe the effect of Dahuang Xiezhuo prescription on the clinical symptoms, blood uric acid, and renal tubular function of patients with immunoglobulin A (IgA) nephropathy in stages 1-2 of chronic kidney disease (CKD) complicated with hyperuricemia (HUA). MethodSixty patients with IgA nephropathy in stages 1-2 of CKD complicated with HUA of spleen and kidney deficiency and combined turbidity and blood stasis syndromes were randomly divided into an observation group and a control group, with 30 cases in each group. The patients in the control group received basic treatment, i.e., losartan potassium tablets 50-100 mg/time, once per day, and sodium bicarbonate tablets 0.5 g/time, three times per day by oral administration, combined with low-salt, low-fat, and low-purine diet. The patients in the observation group received Dahuang Xiezhuo prescription on the basis of basic treatment, one dose per day, twice a day in the morning and evening with warm water. Both groups were treated for two months. The total scores of traditional Chinese medicine(TCM)syndrome, blood pressure, 24 h urinary protein (24 h UTP), blood urea nitrogen (BUN), serum creatinine (SCr) [glomerular filtration rate (eGFR) was calculated by CKD-epidemiology collaboration (CKD-EPI) formula], serum uric acid (SUA), and renal tubular function indexes [urinary α1-microglobulin (α1-MG), urinary β2-microglobulin (β2-MG), urinary kidney injury molecule-1 (KIM-1), and neutrophil gelatinase-associated lipocalin (NGAL)] of the two groups before treatment and two months after treatment were recorded. The clinical efficacy of the two groups was evaluated two months after treatment. ResultAfter 2 months of treatment,the total effective rate in the observation group was 81.48%(22/27),higher than 50.00%(14/28) in the control group(χ2 =6.661,P<0.05). The total scores of TCM syndrome, 24 h UTP, and SUA in the observation group and the observation group were lower than those before treatment (P<0.05), and compared with the control group after treatment, the observation group decreased more significantly (P<0.05). After treatment, the blood pressure in the observation group and the observation group was lower than that before treatment (P<0.05), and there was no significant difference in blood pressure between the two groups after treatment. After treatment, the levels of urinary α1-MG, β2-MG, KIM-1, and NGAL in the two groups were lower than those before treatment (P<0.05), and the observation group was lower than the control group after treatment (P<0.05). There were no significant inter-group and intra-group differences in BUN, SCr, and eGFR levels before and after treatment. There were no obvious abnormalities in blood routine, liver function, and electrolytes before and after treatment in the two groups, and no adverse reactions such as allergies occurred. ConclusionDahuang Xiezhuo prescription can effectively improve the clinical symptoms of IgA nephropathy with HUA (CKD1-2) patients with spleen and kidney deficiency and combined turbidity and blood stasis syndromes, reduce blood uric acid level, alleviate renal tubular injury, and protect the kidney. The curative effect is better than that of basic treatment. 
		                        		
		                        		
		                        		
		                        	
6.A nested case-control study on association between self-reported occupational sulfur dioxide exposure and hypertension
Guoxiu SHI ; Li ZHANG ; Yanli LIU ; Xiaofei ZHANG ; Kang LYU ; Qin SHI ; Chun YIN ; Feng KANG ; Yana BAI ; Shan ZHENG
Journal of Environmental and Occupational Medicine 2022;39(8):856-862
		                        		
		                        			
		                        			Background Current evidence on whether occupational sulfur dioxide (SO2) exposure affects the risk of hypertension is still limited, and the research results of the effect of environmental SO2 exposure on risk of hypertension remain inconsistent. Objective To analyze the association between self-reported occupational exposure to SO2 and the risk of hypertension, and the potential dose-response relationship between the years of exposure to SO2 and the risk of hypertension. Methods Based on the Jinchang cohort, a nested case-control study design was adopted. A total of 841 newly diagnosed hypertension patients were followed up as the case group, and the control group was selected with 1∶1 individual matching based on non-occupational factors and occupational factors, respectively. The former matching conditions included age ±2 years old, same gender, working age ±2 years, and home address in the same sub-district. The latter was limited to working in the same workshop on the basis of the former conditions. Finally, the former included 717 controls and the latter included 488 controls. A unified questionnaire was used to collect general demographic characteristics, lifestyle habits, history of diabetes, family history of hypertension, and information on occupational exposure to SO2 (self-reported history of occupational exposure to SO2 and years of exposure to SO2). Conditional logistic regression model was used to analyze the association between occupational exposure to SO2 and hypertension, and the dose-response relationship between the years of SO2 exposure and the risk of hypertension. Results In the nested case-control study matching with the non-occupational factors, the OR of hypertension in workers with self-reported occupational exposure to SO2 was 2.39 (95%CI: 1.68-3.39); while when matching with the occupational factors, the OR of hypertension in workers with self-reported occupational exposure to SO2 was 1.48 (95%CI: 1.04-2.12). The results of the dose-response relationship showed that as the SO2 exposure years increased from 1-9 years, 10-19 years, 20-29 years, and 30 years and above, in the nested case-control study matching with non-occupational factors, the ORs of hypertension were 1.85 (95%CI: 0.68-5.08), 1.46 (95%CI: 0.58-3.67), 1.64 (95%CI: 1.00-2.67), and 4.95 (95%CI: 2.63-9.31), respectively; in the nested case-control study matching with occupational factors, the ORs of hypertension were 0.98 (95%CI: 0.40-2.41), 1.84 (95%CI: 0.72-4.70), 1.37 (95%CI: 0.82-2.29), and 2.44 (95%CI: 1.37-4.35), respectively. The two dose-response relationships were positive by χ2 trend test (Ptrend<0.05). Conclusion Self-reported occupational exposure to SO2 is associated with the risk of hypertension in the study population, and the hypertension risk increases with the increase of SO2 exposure years.
		                        		
		                        		
		                        		
		                        	
7.Single Cell Traction Force Measured by Foldable Microplates
Lijun ZHAO ; Chenyan WANG ; Quanyou ZHANG ; Di HUANG ; Jinchuan HOU ; Weiy CHEN
Journal of Medical Biomechanics 2022;37(2):E287-E291
		                        		
		                        			
		                        			 Objective To fabricate a foldable microplate for single cell culture and establish finite element model of the folding microplate, so as to calculate traction force of single cells during contraction in three-dimensional (3D) state.Methods The folding angle of the microplate casued by cell traction force was calculated. Then the relation between bending moment and folding angle as well as the relation between traction force and bending moment were derived by using finite element simulation, so as to realize the characterization of traction force for singel cell in 3D state.Results The folding angles of the microplate with HSF and MC3T3-E1 cells in 3D state were 73°-173° and 49°-138°, respectively. The single cell traction forces of HSF and MC3T3-E1 cells were 55-210 nN and 52-161 nN, respectively.Conclusions The proposed method for measuring traction force of single cells in 3D state by fabricating the foldable microplate for single cell culture will provide some references for further development of calculating traction forces in 3D cell adhesion, spreading and migration. 
		                        		
		                        		
		                        		
		                        	
8.Modified Shengjiangsan Inhibits Necroptosis by Mediating RIP1/RIP3/MLKL Signaling Pathway and Reduces Renal Fibrosis in Rats with Diabetic Nephropathy
Ruijing SONG ; Xinxin ZHANG ; Fei GAO ; Miao TAN ; Jinchuan TAN
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(17):33-42
		                        		
		                        			
		                        			ObjectiveTo observe the mechanism of modified Shengjiangsan in necroptosis and renal fibrosis of rats with diabetic nephropathy based on receptor-interacting protein (RIP)1/RIP3/mixed lineage kinase domain-like protein (MLKL) signaling pathway. MethodSeventy-five SD rats were randomly divided into a model group, a normal group, three high, medium, and low-dose modified Shengjiangsan groups (4.365, 8.73, 17.46 g·kg-1), and an irbesartan group (0.013 5 g·kg-1). After 4 weeks of intragastric administration, the levels of 24 h urine protein (UTP), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) of rats in each group were determined, as well as the changes in degree of renal pathology. Real-time quantitative polymerase chain reaction (Real-time PCR) and immunohistochemistry were used to detect the mRNA and protein expression levels of IL-1β, TNF-α, monocyte chemoattractant protein-1 (MCP-1), transforming growth factor-β1 (TGF-β1), and nuclear factor kappa B (NF-κB) in kidney tissues of rats. Western blot assay was used to detect the expression levels of key proteins in the RIP1/RIP3/MLKL signaling pathway. ResultAs compared with the normal group, the renal interstitial fibrosis in the model group was obvious, and the 24 h UTP, IL-1β, TNF-α levels were significantly increased (P<0.05). In the model group, the mRNA and protein expression levels of IL-1β, TNF-α, MCP -1, TGF-β1, and NF-κB in the kidney tissues were significantly increased (P<0.05), and protein expression levels of RIP1, RIP3, p-MLKL, and MLKL were significantly increased (P<0.05). Compared with the model group, all modified Shengjiangsan groups and the irbesartan group improved the levels of renal interstitial fibrosis in rats to varying degrees. As compared with the model group, the 24 h UTP levels in all modified Shengjiangsan groups and the irbesartan group were decreased to varying degrees (P<0.05), the content of IL-1β and TNF-α in the serum were decreased (P<0.05), the mRNA and protein expression levels of IL-1β, TNF-α, MCP-1, TGF-β1, and NF-κB in renal tissues was down-regulated (P<0.05), and the protein expression levels of RIP1, RIP3, p-MLKL, and MLKL were down-regulated (P<0.05). ConclusionModified Shengjiangsan ameliorates renal injury of rats with diabetic nephropathy, and the mechanism may be related to the down-regulation of the RIP1/RIP3/MLKL signaling pathway, the prevention of renal tissue necroptosis, and the inhibition of renal fibrosis. 
		                        		
		                        		
		                        		
		                        	
9.The Combined Effect of Dyslipidemia on the Incidence of Type 2 Diabetes: A Prospective Cohort Study in Northwest of China.
Min Zhen WANG ; Tian DAI ; Shan ZHENG ; Cheng YU ; Miao XIA ; Hong Yan YANG ; De Sheng ZHANG ; Chun YIN ; Ya Fei JIN ; Ning CHENG ; Ya Na BAI
Biomedical and Environmental Sciences 2021;34(10):814-818
10.Effects of Outdoor Temperature on Blood Pressure in a Prospective Cohort of Northwest China.
Shan ZHENG ; Min Zhen WANG ; Zhi Yuan CHENG ; Feng KANG ; Yong Hong NIE ; Xiu Ying MI ; Hai Yan LI ; Lan JIN ; Ya Wei ZHANG ; Ya Na BAI
Biomedical and Environmental Sciences 2021;34(2):89-100
		                        		
		                        			Objective:
		                        			The relationship between outdoor temperature and blood pressure (BP) has been inconclusive. We analyzed data from a prospective cohort study in northwestern China to investigate the effect of outdoor temperature on BP and effect modification by season.
		                        		
		                        			Methods:
		                        			A total of 32,710 individuals who participated in both the baseline survey and the first follow-up in 2011-2015 were included in the study. A linear mixed-effect model and generalized additive mixed model (GAMM) were applied to estimate the association between outdoor temperature and BP after adjusting for confounding variables.
		                        		
		                        			Results:
		                        			The mean differences in systolic blood pressure (SBP) and diastolic blood pressure (DBP) between summer and winter were 3.5 mmHg and 2.75 mmHg, respectively. After adjusting for individual characteristics, meteorological factors and air pollutants, a significant increase in SBP and DBP was observed for lag 06 day and lag 04 day, a 0.28 mmHg (95% 
		                        		
		                        			Conclusions
		                        			This study demonstrated a significant negative association between outdoor temperature and BP in a high-altitude environment of northwest China. Moreover, BP showed a significant seasonal variation. The association between BP and temperature differed by season and individuals' demographic characteristics (age, gender, BMI), unhealthy behaviors (smoking and alcohol consumption), and chronic disease status (CVDs, hypertension, and diabetes).
		                        		
		                        		
		                        		
		                        			Adult
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		                        			Blood Pressure/physiology*
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		                        			China/epidemiology*
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		                        			Environmental Exposure/statistics & numerical data*
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		                        			Female
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		                        			Humans
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		                        			Male
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		                        			Middle Aged
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		                        			Prospective Studies
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		                        			Risk Factors
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		                        			Seasons
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		                        			Temperature
		                        			
		                        		
		                        	
            
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