Objective To study the effects of high matrix metalloproteinase 9 (MMP 9 ) on the biological behaviors of fibroblasts,in order to clarify the possible mechanisms in the wound healing of diabetic foot.Methods Establish the cell model of skin fibroblast with high expression of MMP 9 by high glucose (22.0 mmol/L) and high homocysteine (100 μmol/L) co-culture.Control group was incubated with normal glucose (5.5 mmol/L).Realtime PCR,ELISA and gelatin zymography were used to detect the MMP 9 mRNA,protein expression and activity of MMP 9.Flow cytometry,CCK-8,ELISA assay,scratch test and transwell were used to detect cell proliferation,viability,collagen (hydroxyproline) secretion,horizontal migration and vertical migration of cells.Results Data were expressed as (x- ± s).Differences were considered significant if their P value was ＜ 0.05.Results The expression of MMP 9 mRNA,protein levels and the activity of MMP 9 in high MMP 9 group were 6.05 folds,4.12 folds and 1.58 folds higher than those in control group (P ＜ 0.01,respectively).The proportion of S phase cells,proliferation index,cell viability,collagen (hydroxyproline) secretion,6 h horizontal migration rate and the number of vertical migration cells in high MMP9 group decreased by 29.8 ％,18.1％,23.3 ％,68.7 ％,45.0 ％ and 21.4 ％ than those in control group (P ＜ 0.01,respectively ). Conclusions Fibroblast with high expression of MMP 9 have decreased proliferation,activity,collagen secretion and reduced migration,which suggests that MMP 9 may inhibit the biological behaviors of fibroblasts.

Objective:To investigate the correlation between body composition and cardiovascular disease (CVD) in patients with chronic kidney disease (CKD).Methods:CKD patients who were hospitalized in the Department of Nephrology of Chongqing General Hospital from January 2017 to December 2019 and had complete clinical biochemical data were divided into CKD patients with CVD and CKD patients without CVD according to their medical history and corresponding auxiliary examinations. Clinical data were collected and anthropometric measurements were conducted. Skeletal muscle index (SMI), appendage lean mass/height 2, total body fat (TBF), visceral adipose tissue (VAT), bone mineral capacity, bone mineral density and et al, were measured by dual-energy X-ray absorptiometry. T test, U test and Chi-square test were used for statistical analysis. Logistic regression was used to analyze the relationship between body composition and CVD. Results:A total of 604 CKD patients were included in this study, including 560 patients (92.7%) with CKD stage 3, 44 patients (7.3%) with CKD stage 4, and 180 CKD patients with CVD (29.8%), 424 CKD patients without CVD (70.2%). Compared with CKD patients without CVD, the proportion of men, the proportion of hypertension, the proportion of diabetes, age, duration of CKD, systolic blood pressure, blood uric acid, waist to hip ratio and waist circumference were higher (all P<0.05), while low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol, and estimated glomerular filtration rate (eGFR) were lower in CKD patients with CVD (all P<0.05). In terms of body composition, SMI ( t=-11.964, P<0.001) and body mass index ( t=-4.462, P<0.001) in CKD patients with CVD were significantly lower than those in CKD patients without CVD, but VAT ( t=3.089, P=0.002) and TBF ( t=5.177, P<0.001) in CKD patients with CVD were significantly higher. After adjusting for confounders such as age, CKD duration, hypertension history, diabetes history, LDL-C, body mass index, eGFR, TBF, etc. by multivariate logistic regression analysis, the risk of CKD patients suffering from CVD increased significantly with the decrease of SMI [with SMI high tertile (36.37%-50.80%) as reference, SMI middle tertile (28.23%-36.31%): OR=1.49, 95% CI 1.24-1.71, P=0.003; SMI low tertile (15.28%-28.19%): OR=2.17, 95% CI 1.79-2.62, P<0.001], and VAT was not found to be associated with the risk of CVD in CKD patients ( P>0.05). Conclusion:Reduction of SMI is independently associated with CVD in CKD patients.

Objective To investigate the effects of S100B on the expressions of dopamine receptors and the synthesis,metabolism of neurotransmitters dopamine,5-hydroxytryptamine which are related to the abnormal motor coordination of Parkinson's disease (PD).Methods The hS100B transgenic mice were established.The mice were divided into S100B transgenic group (TG,n =14),S100B knockout group (KG,n =14) and the non-transgenic control group (CG,n =14).The motor coordination ability of mice was measured by the Rota-rod test.The expressions of dopamine D1 receptor (D1DR),dopamine D2 receptor (D2DR),tyrosine hydroxylase (TH) and phosphorylated TH at Ser19,Ser31,Ser40 in brain tissue were detected by reverse transcription polymerase chain reaction and Western blotting.The levels of Tyr,levodopa,dopamine,homovanillic acid,Trp,5-hydroxytryptamine and 5-hydroxyindoleacetic acid in mesencephalon were measured by high performance liquid chromatography with fluorescence detection.Results Compared with CG,the motor coordination ability of mice (s) showed progressive decline in TG (3 months:4.60±0.30vs4.25±0.21,q =5.194;6 months:4.52±0.31 vs4.07±0.22,q =6.139;9 months:4.43 ± 0.25 vs 3.60 ± 0.18,q =13.484;all P ＜ 0.05),the expressions of D2DR mRNA and protein decreased (1.34 ± 0.13 vs 0.48 ± 0.07,q =21.578;1.05 ± 0.15 vs 0.69 ± 0.10,q =8.063,both P＜0.05) and phosphorylated TH at Serl9 and Ser40 increased (0.95 ±0.10 vs 1.14-0.13,q =4.972;0.94 ± 0.12 vs 1.17 ± 0.14,q=5.382,both P＜ 0.05),the levels of levodopa,dopamine and homovanillic acid were elevated (87.04 ± 11.77 vs 115.28 ± 16.80,q =4.764;56.66 ± 9.87 vs 72.96 ± 11.02,q=3.923;26.58 ± 8.11 vs 38.65 ± 6.67,q=3.981,all P＜ 0.05),the leve1 of 5-hydroxytryptamine was reduced (925.50 ± 74.26 vs 637.87 ± 56.76,q =11.084,P ＜ 0.05),the ratios of homovanillic acid/dopamine and 5-hydroxyindoleacetic acid/5-hydroxytryptamine increased (0.45 ± 0.05 vs 0.54±0.08,q =3.325;0.94±0.07 vs 1.42±0.12,q =12.367,both P＜0.05) in the brain of TG at the age of 9 months old.There was no significant difference of detection indexes between KG and CG.Conclusions S100B plays an important role in the development of PD and the brain-specific S100B transgenic mice can be used to investigate the function of S100B gene on the development of PD.

Results There was no significant difference in the distribution of genotypes and allele frequencies of HSP 70-1 and HSP70-hom between case group and control group (P>0.05), but there was significant difference in the distribution of genotypes and allele frequencies of HSP70-2 between the two groups(P<0.05).There was no significant difference in the distribution of genotypes and allele frequencies of HSP70-1,HSP70-2 and HSP70-hom between Hui and Han people (P>0.05)or in the distribution of genotypes of HSP 70-2 between males and females in case group, but there was significant difference in the distribution of allele frequencies of HSP 70-2 in case group .There was no significant difference in the distribution of genotypes and allele frequencies of HSP 70-1 and HSP70-hom between males and females in case or control group .Logistic analysis confirmed that the WC, TG, TC, LDL-C, SBP, family history of diabetes and G allele were risk factors of T 2DM. Conclusion GG genotype and G allele of HSP 70-2 (+1267 ) SNP may be genetic markers for the susceptibility of type 2 diabetes.There is no significant difference in the distribution of genotypes and allele frequencies of HSP 70-1,HSP70-2 and HSP70-hom between Hui and Han nationalities .Family history of diabetes, LDL-C, TC, and G allele of HSP70-2 (+1267) SNP are the main risk factors of T2DM while HDL-C is a protective factor.

BACKGROUND:The high level of matrix metalloproteinase 9 (MMP9) is thought to slow down the healing of diabetic foot ulcers. Whether it can influence the biological behaviors of skin fibroblasts and affect wound healing is stillunclear. The present study aimed to observe changes in the biological behaviors of rat dermal fibroblasts induced by high expression of MMP9 and to clarify the possible mechanisms of wound healing for diabetic foot. METHODS:A cellmodel of skin fibroblast with high expression of MMP9 was established by co-culture of high glucose (22.0 mmol/L) and homocysteine (100 μmol/L). A control group was incubated with normal glucose (5.5 mmol/L). Realtime PCR, ELISA and gelatin zymography were used to detect the MMP9 mRNA, protein expression and activity of MMP9. Flow cytometry, CCK-8, ELISA assay, scratch test and transwellwere used to detect cellproliferation, viability, collagen (hydroxyproline) secretion, horizontal migration and vertical migration of cells. The data were expressed as mean±SD. P value less than 0.05 was considered statistically significant. RESULTS:The expression of MMP9 mRNA, protein levels and the activity of MMP9 were much higher in the high MMP9 group than in the control group (7.05±1.02 vs. 1.00±0.00, 206.9±33.6 pg/mL vs. 40.4±5.9 pg/mL, and 1.47±0.13 vs. 0.57±0.12, respectively,P<0.01). The proportion of S-phase cells, proliferation index, cellviability, collagen (hydroxyproline) secretion, horizontal migration rate and the number of vertical migration cells were lower in the high MMP9 group than in the control group (P<0.01). CONCLUSION:Fibroblasts with a high expression of MMP9 decreased proliferation, activity, secretion and migration of collagens, suggesting that MMP9 may inhibit the biological behaviors of fibroblasts.

Objective To describe the clinical characteristics of a patient with Gitelman syndrome,and to identify the associated SLC12A3 gene mutations.Methods A suspected case of teenager-onset Gitelman syndrome was observed in our hospital.It was further confirmed by clinical manifestations and auxiliary examination.In addition,direct sequencing for the exons of SLC12A3 gene and CLCNKB gene region was conducted to identify the probable disease-associated mutations.Results The case showed characteristics of hypokalemia,hypomagnesemia,and low level of urinary calcium and onset by age of 18.By excluding the possibilities of long-term use of thiazide diuretics,laxatives,chronic vomiting and diarrhea,he was finally diagnosed as a case of Gitelman syndrome.Furthermore,by Sanger direct sequencing,2 coding variations were identified in SLC12A3 gene region,including T304M and L488P.L488P was a new heterozygous mutation.Conclusion Detection of SLC12A3 gene mutation could facilitate the diagnosis of Gitelman syndrome and improve prognosis.

Heroin can be metabolized easily in body and the mail metabolites are 6-MAM, morphine and so on. At present, there are urine, blood, hair and so on as specimens for detection, while the analytical technology conclude TLC, GC, HPLC, GC/MS, LC/MS, IA, CE etc. In this paper, these technologies used for heroin's metabolites were viewed in order to provide some reference to the study in relative field.
Chromatography, High Pressure Liquid ; Forensic Medicine ; Gas Chromatography-Mass Spectrometry ; Hair/chemistry* ; Heroin/metabolism* ; Heroin Dependence/metabolism* ; Humans ; Morphine/analysis* ; Morphine Derivatives/analysis* ; Substance Abuse Detection/methods*

OBJECTIVE: To confirm whether formaldehyde disturb detecting carbon monoxide in blood. To give an evidence that can be used for detecting carboxyhemoglobin more accurately in carbon monoxide posioning appraises. METHODS: Blood samples came from carbon monoxide poisoning and the health were collected. Regular methods for detecting carboxyhemoglobin were used. Observing and comparing the detection results between which were spiked with methanal and no spiked one were performed. RESULTS: Methanal will affect the result of following experiments such as heating, adding NaOH, absorbed by PdCl2 and spectrophotometry. CONCLUSION The samples which contaminated by formaldehyde couldn't be used for detecting carboxyhemoglobin.
Carbon Monoxide/blood* ; Carbon Monoxide Poisoning/diagnosis* ; Carboxyhemoglobin/analysis* ; Forensic Medicine ; Formaldehyde/pharmacology* ; Humans ; Spectrophotometry/methods* ; Temperature

Objective To study the level of serum lipids and body-fat content of high-fat diet induced pbesity rats(DIO).explore the relationship between intracellular caleium and ventrieufar arrhythmia.Methods Sixty male Sprague-Dawley(SD)rats were divided into control group(15)and experiment group(45),high-fat diet was administrated for 12 weeks to established obesity model,15 rats were selected into obesity group according to their body weight gain.The standard 2-lead electrocardiograph was used to detect the incidence and scores of arrhythmia induced by barium chloride(BaCl2,0.1 mg/kg)for 1 hour on every 8 rats from different groups respectively.Body-fat content.the level of serum total cholesterol(TC),triglyceride(TG),low density lipopmtein cholesterol (LDL-C) and high density lipopmtein cholesterol (HDL-C)were measured.The epididymal(EP),retroperitoneal (RE) and mesenteric(ME)white adipose pads was measured to obtain the body fat content.Single ventricular myrocytes of rats were isolated by enzymatic dissociation.The confocal laser scanning microscope was used to record basic intraceUular calcium level([Ca2+]i).Results The body-fat content in obesity group[(7.71±0.74)%] was significantly higher than control group[(4.69±0.37)%](t＝3.650,P＜0.05).The level of serum TC,TGand LDL-C were significantly higher(t＝3.801,2.778,3.536.P＜0.05) in obesity group[(1.26±0.04),(0.58±0.10),(0.51±0.04)mmol/L]than those in control group[(0.92±0.08),(0.29+0.03),(0.31±0.04) mmol/L].The level of serum HDL-C wa8 decreased gradually from control group[(0.53±0.05)mmol/L] toobesity group[(0.52±0.02)mmol/L],but there waft no significant difference between them(t＝0.186,P＞0.05).The incidence of arrhythmia induced by BaCl2(0.1 mg/kg)in obesity group was significantly higher than controlgroup(X2＝5.333,P＜0.05),and the scores of arrhythmla was increased in obesity group(2.5±0.6)too.The fluorescence intensity standing for[Ca2+]i was increased significantly(t＝2.409,P＜0.05)from obesity group(247.96±20.03)to control group (174.25±23.13).Conclusion As the free cytosolic calcium begin to accumulate,the arrhythmia morbidity is increased in obesity rats.

Primary angiitis of the central nervous system is a rare and difficult entity. Here we represented the clinical and pathological features of a patient with little response to steroid before definite diagnosis. The 50-year-old male had a fluctuating disease course for more than 3 years. He presented visual disorders, seizure, cognitive impairment, hypersomnia, unsteady gait, dysphasia, dysphagia, and incontinence. Magnetic resonance imaging showed multiple, supratentorial and infratentorial abnormal signals, while cerebrospinal fluid and cerebral angiography were normal. Magnetic resonance spectrum showed a decrease of N-acetyl-aspartate. Brain biopsy revealed nongranulomatous lymphatic vasculitis with reactive gliosis, cicatrization, demyelination and focal hemorrhages.
Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Vasculitis, Central Nervous System ; diagnosis