OBJECTIVETo investigate the acute effects of aldosterone (ALD) on mesenteric resistance vessels in normal or heart failure (HF) rats and its mechanism.

METHODSHF model was adopted by in vivo ligation of left anterior descending coronary artery in SD rats; segments of third-order branches of mesenteric artery were isolated and dissected into about 2 mm rings for isometric force recording.

RESULTSPretreated with ALD for 10 min,phenylephrine (PE)-induced contraction of normal mesenteric artery decreased first and then increased compared to control group along with the increase of the concentration of PE while decreased in HF rats. This effect was attenuated by ALD receptor-special antagonist eplerenone partially. ALD increased Ach-induced endothelial-dependent vascular relaxation significantly compared to control group both in normal and HF rats. Pretreated with ALD and dexamethasone (DEX) for 10 min, the effects of ALD on PE-induced contraction were weakened in mesenteric artery both of normal and HF rats. And this reaction of DEX to ALD-treated mesenteric in normal rats was attenuated by RU486 partially.

CONCLUSIONALD has biphasic effect in PE-induced response on mesenteric artery of normal rats, while reduces the sensitivity of mesenteric artery to PE in HF rats. DEX attenuates the biphasic effect of ALD on artery of normal rat partially but has no significant effect on that of HF rats.

Aldosterone ; pharmacology ; Animals ; Heart Failure ; physiopathology ; Male ; Mesenteric Arteries ; drug effects ; physiology ; Phenylephrine ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Vasoconstriction ; drug effects

Objective To investigate the expression and aberrant methylation of Ptchl gene in hedgehog signal pathway in carcinogenesis of human gastric cancer.Methods The total RNA and genomic DNA were extracted from 10 human gastric carcinoma tissues,adjacent tissues(＞3 cm from cancerous tissue)and gastric cancer eell line AGS.Ptchl mRNA expression was detected by real-time quantitative reverse transcription PCR.The pattern of CpG island in Ptchl gene 5'regulation sequence was analyzed by software and its methylation extent was tested by bisulfite sequencing PCR.Results The analysis of CpG island(starting-3950 bases upstream of the Ptchl mRNAla transcription start site and ending 2050 bases downstream)revealed that there were two CpG islands in Ptchl gene 5' regulation sequence(first CpG:-1139 bp～+860 bp;second CpG:+875 bp～+1692 bp).Bisulfite sequencing PCR analysis of 19 CpG sites included in the first CpG island(-870 bp～+229 bp)showed that there was methylation present in all cell lines and the average extent of the methylation of these CpG sites was significantly higher in cancerous tissues(64%±32%,ranged 16%～100%)than that in adjacent tissues(13%±14%,ranged 0%～42%,P＜0.05).There was a negative correlation of the Ptchl methylation with its expression.Conclusion The high methylation of Ptchl gene that involves in the carcinogenesis of human gastric carcer will be a new biomarker for gastric carcer.

OBJECTIVE: To identify pathogenic variants in 5 sporadic patients and two Chinese pedigrees affected with 17-hydroxylase deficiency (17-OHD). METHODS: Peripheral blood samples were collected with informed consent. Variants of CYP17A1 gene were screened by PCR and Sanger sequencing. Suspected mutations were validated in other members of the pedigrees. RESULTS: Gene sequencing has identified a homozygous c.985_987delTACinsAA (Y329Kfs) mutation in exon 6 of the CYP17A1 gene in 4 patients and the sister of case 3. Case 1 was found to harbor compound heterozygous mutations c.1459_1467del9 (p.D487_F489del) and c.1244-3C>A. The parents and brother of cases 2 and 5 were heterozygous carriers of a c.985_987delTACinsAA(Y329Kfs) mutation. CONCLUSION Mutations of the CYP17A1 gene probably underlie the pathogenesis of 17-OHD, for which c.985_987delTACinsAA(Y329Kfs) is the most common. The c.1244-3C>A is a novel mutation. Above results have facilitated genetic counseling for the affected families.
Adrenal Hyperplasia, Congenital ; genetics ; Exons ; Female ; Humans ; Male ; Mutation ; Pedigree ; Steroid 17-alpha-Hydroxylase ; genetics

Objective: To establish the immortalized mouse brain microvascular pericytes model and to apply to the cerebrovascular toxicants screening study. Methods: Brain pericytes were isolated from 3 weeks of mice by tissue digestion. Immortalized pericyte cell line was constructed by infecting with LT retrovirus. Monoclone was selected to purify the immortalized pericyte cell line. The pericyte characteristics and purity were explored by immunocytochemistry. Cell proliferation was measured by using the Pomega MTS cell Proliferation Colorimetric Assay Kit. Pericytes were treated with 0, 160, 320, 640, 1 280, 2 560 μmol/L lead acetate, 0, 5, 10, 20, 40, 80 μmol/L cadmium chloride and 0, 5, 10, 20, 40, 80 μmol/L sodium arsenite in 24 hours. Cell toxicity of each group was determined by MTS assay, median lethal dose (LD₅₀) was calculated in linear regression. Results: Mouse brain pericytes were successfully isolated by tissue separation and enzyme digestion method. After immortalized by LT retroviruses, monoclone was selected and expanded to establish pericyte cell line. The brain pericytes exhibited typical long spindle morphology and positive staining for α-SMA and Vimentin. The proliferation of brain pericytes cell lines was very slowly, and the doubling time was about 48 hours. The proliferation of immortalized brain pericytes cell lines was very quickly, and the doubling time was about 24 hours. After lead acetate, cadmium chloride and sodium arsenite treatment for 24 hours respectively, gradual declines in cell viability were observed. The LD₅₀ of lead acetate was 2 025.0 μmol/L, the LD₅₀ of cadmium chloride was 36.6 μmol/L, and the LD₅₀ of sodium arsenite was 33.2 μmol/L. Conclusion The immortalized mouse brain microvascular pericyte model is established successfully by infecting with LT retrovirus, and can be applied to screen cerebrovascular toxicants. The toxicity of these toxicants to immortalized mouse brain microvascular pericyte is in sequence: sodium arsenite,cadmium chloride, lead acetate.

Objective: To investigate the effects of cerebral cavernous malformation 3 (CCM3) gene knockout on the lead exposure-induced blood-brain barrier malfunction in mice brain, and the relationship between CCM3 knockout and the Alzheimer's disease (AD). Methods: Wide type (WT) mice and CCM3⁺/- mice were divided into 4 groups, control group and lead exposed group in WT as well as CCM3^+/- mice. Lead exposed groups were treated with 0.05% lead acetate in drinking water for 12 weeks, while control group drink deionized water freely. Blood lead and brain lead levels in each group were detected by graphite furnace atomic absorption spectrometry. The brain tissue of each group was made into paraffin sections, whose morphology were observed by HE staining. The expression of Aβ_1-42 in brain tissue was detected by immunohistochemistry and the brain capillaries were labeled by VRGFR2. The protein expression of Claudin-5, ZO-1, and p-Tau was detected by Western blot. The brain tissue RNA was extracted and the relative expression of LRP-1 mRNA was detected by qRT-PCR. Results: The levels of blood lead WT (216.07±84.16) and CCM3^+/- (189.64±101.86) μg/L in lead exposed group were higher than those in control group WT (19.52±11.46) and CCM3^+/- (11.79±8.20) μg/L, the difference was statistically significant (t=4.18, P=0.006; t=3.79, P=0.016). The levels of brain lead WT (1.78±0.69) and CCM3^+/- (1.74±0.66) μg/L were higher than those in control group WT (1.06±0.87) and CCM3^+/- (0.97±0.64) μg/L, the difference was statistically significant (t=3.67, P=0.018; t=3.88, P=0.015). The HE staining showed no obvious lesions in the brain of each group of mice. The results of immunohistochemistry showed that there was no Aβ₁-42 deposition in the brain of mice in each group. The numbers of microvessels in the brain of CCM3^+/- mice in the lead exposed group were decreased. Compared with the relative expression levels of Claudin-5 (WT: 1.30±0.03, CCM3^+/-: 1.07±0.08) in control group mice brain, the relative expression of Claudin-5 (WT: 0.96±0.04, CCM3^+/-: 0.59±0.01) was decreased with statistical significance (F=199.27, P<0.001). The relative expression level of LRP-1 gene mRNA in brain of lead exposed group (WT: 0.32±0.10, CCM3^+/-: 0.06±0.01) was higher than that of unexposed group (WT:1.00±0.06, CCM3^+/-:2.12±0.18), the difference was statistically significant (F=288.29, P<0.001). The relative expression level of LRP-1 gene mRNA in brain of CCM3⁺/- mice exposed to lead was lower than that of WT mice ((0.06±0.01)vs(0.32±0.10), t=26.90, P<0.001). Conclusion The mice did not show significant AD-like lesions under low-does lead exposure, but resulted in early damage of brain blood-brain barrier and early changes of AD-like lesions in mice, with CCM3^+/- mice being sensitive to lead exposure stronger than that of WT mice, suggesting that deletion of CCM3 gene may be one of the potential risk factors for accelerating the development of AD in mice exposed to lead.

Objective:To explore the correlation between dietary fiber intake and intestinal barrier function in patients with irritable bowel syndrome with diarrhea (IBS-D).Methods:IBS-D patients were recruited from May 2019 to October 2019 at the clinic of gastroenterology department in China-Japan Friendship Hospital and healthy controls (HCs) were recruited by advertisement. Clinical manifestations, psychological status and quality of life were assessed using standardized questionnaires. A food frequency questionnaire was used to assess dietary habits in the preceding year. Serum diamine oxidase (DAO) was measured via ELISA.Results:64 IBS-D patients and 35 HCs were enrolled, with no significant difference in sex ratio, age and BMI between the two groups. Second to health concern, food avoidance was the dominant impacting factor for quality of life in IBS-D patients. The intake frequency of dietary fiber was decreased in IBS-D patients, and the intake frequencies of dietary fiber-rich foods were significantly lower in IBS-D patients ( P < 0.01 for tubers, P = 0.002 for vegetables, P = 0.019 for fruits and P = 0.045 for legumes). On the other hand, the intake frequencies of processed meat ( P < 0.01), greasy food ( P = 0.009), barbecued food ( P = 0.002) and animal offal ( P = 0.003) were significantly higher in IBS-D patients compared with HCs, indicating the increased intake frequency of fat. Multivariate logistic regression showed that tubers might reduce the risk of IBS-D ( OR = 0.409,95% CI: 0.232 to 0.722, P = 0.002). The frequency of abdominal pain was positively associated with the intake frequency of greasy food in IBS-D patients. Serum DAO was measured in 37 IBS-D patients and 27 HCs. IBS-D patients had significantly higher serum DAO than HCs ( 77.0 [55.3, 100.6] μg/L vs 42.5 [28.0, 58.2] μg/L, P < 0.01). Among all the participants with serum DAO test results, the level of DAO was negatively associated with the intake frequencies of tubers, vegetables and fruits while positively associated with the intake frequencies of processed meat and barbecued food. Conclusions:Food avoidance was an important impacting factor for quality of life in IBS-D patients. IBS-D patients might have insufficient dietary fiber intake and excessive fat intake. Tubers could possibly reduce the risk of IBS-D. The decreased intake frequency of dietary fiber might have a role in intestinal barrier dysfunction in IBS-D patients.

Objective To investigate the effects of anti-HLA donor-specific antibodies(DSA)and desensitization for DSA+patients on engraftment of haploidentical hematopoietic stem cell transplantation(haplo-HSCT).Methods The patients who underwent haplo-HSCT and examinations for HLA antibodies and DSA in our department from March 2017 to July 2023 were recruited in this study.The effects of desensitization measure on engraftment in the DSA+patients after haplo-HSCT were analyzed.Results Among the 70 patients who underwent haplo-HSCT and test for HLA antibodies,15(21.4％)patients were DSA positive,including 7(46.7％)cases of strong positive,3(20.0％)cases of moderate positive,and 5(33.3％)cases of weak positive.The median duration for neutrophil implantation was significantly extended in the DSA+patients than the negative patients(P=0.027).For the 6 patients developed graft failure(GF),4 were DSA+which was statistically higher than the DSA-patients(P=0.025).Multivariate regression analysis showed that DSA was an independent factor affecting GF(HR=9.273,95％CI:1.505～57.124,P=0.016).Among the 10 patients(7 strong positive and 3 moderate positive DSA)received desensitization therapy,4 patients received combination desensitization,with a 100％rate of successful transplantation,and 6 received single desensitization,with 4(66.7％)experiencing GF,so the GF rate was obviously lower in the combination than the single desensitization(P=0.008).Conclusion In haplo-HSCT patients,DSA is an important factor leading to implantation delay and GF.While,single desensitization treatment has limited efficacy.In combined DSA desensitization therapy,the decrease of antibody titer should be dynamically monitored to ensure the successful implantation of stem cells and reduce GF rate.

Objective To explore the relationship between non-high density lipoprotein cholesterol(non-HDL-C)level and leptomeningeal collateral circulation in patients with acute middle cerebral artery occlusion.Methods A total of 85 patients with first-onset acute cerebral infarction with middle cerebral artery M1 segment occlusion were enrolled.According to the results of DSA,LMC circulation was assessed by American Society of Interventional and Therapeutic Neuroradiology/Society of Interventional Radiology Collateral Circulation Assess-ment System.All patients were assigned to better LMC circulation group(score 2～4,n = 30)and worse LMC circulation group(score 0～1,n = 55),and the levels of non-HDL-C were compared between the two groups.Results The levels of LDL-C and non-HDL-C in worse LMC circulation group were significantly higher than those of the better LMC circulation group(P = 0.026,P = 0.010).non-HDL-C was an independent risk factor for the worse LMC circulation(OR = 3.019,95%CI:1.053～8.658,P = 0.04).LMC circulatory score of patients was negatively correlated with the levels of non-HDL-C level(r =-0.228,P = 0.036).The AUC of non-HDL-C predicted for the worse LMC circulation was 0.638(95%CI:0.521～0.755,P = 0.036).Conclusions non-HDL-C in patients with acute cerebral infarction was significantly related to worse LMC circulation,and was a risk factor for worse LMC circulation.It is suggested that the higher expression of non-HDL-C could be used to predict worse LMC circulation as a serological indicator.

OBJECTIVE: To explore the feasibility of radical resection for cancer patients complicated with coronavirus disease 2019 (COVID-19). METHODS: The management and clinical outcome of a sigmoid cancer patient with COVID-19 were analyzed. RESULTS: The inflammation indicators and fever of this patient were effectively controlled and the lung lesions remained stable after active anti-viral treatment, then the radical colorectomy was performed after the viral negative conversion for twice. CONCLUSIONS The case indicates that radical resection can be performed in SARS-CoV-2 patients with twice-negative SARS-CoV-2 nucleic acid testing results.
Betacoronavirus ; isolation & purification ; Colonic Neoplasms ; complications ; surgery ; Coronavirus Infections ; complications ; therapy ; Disease Management ; Humans ; Pandemics ; Pneumonia, Viral ; complications ; therapy ; Treatment Outcome

OBJECTIVE: To explore the feasibility of surgical treatment for cancer patients complicated with corona virus disease 2019 (COVID-19). METHODS: The management and clinical outcome of a sigmoid cancer patient with COVID-19 were analyzed. RESULTS: The inflammation indicators and fever of this patient were effectively controlled and the lung lesions remained stable after active anti-viral treatment, then the radical colorectomy was performed after the viral negative conversion for twice. CONCLUSIONS The case indicates that it may feasible to undergo radical tumor surgery for cancer patients with COVID-19 after the virus nucleic acid testing turns negative and more studies are needed to confirm this conclusion.
Antiviral Agents ; therapeutic use ; Betacoronavirus ; Clinical Laboratory Techniques ; Colonic Neoplasms ; complications ; surgery ; virology ; Coronavirus Infections ; complications ; diagnosis ; drug therapy ; Fever ; Humans ; Nucleic Acid Amplification Techniques ; Pandemics ; Pneumonia, Viral ; complications ; diagnosis ; drug therapy