Zofenopril antitumor activity in mice bearing Ehrlich solid carcinoma: Modulation of PI3K/AKT signaling pathway

Angiotensin-II (AgII) is thought to be crucial for tumor growth and progression. Moreover, hydrogen sulfide (H2S) performs a controversial action in cancer pathology. Zofenopril (ZF) is an angiotensin-converting enzyme (ACE) inhibitor with H2S donating properties. Hence, this study aims at investigating the tumor suppressor activity of ZF and elucidating the involved trajectories in Ehrlich's solid tumor (EST)-bearing mice. EST was induced by the intradermal injection of Ehrlich's ascites carcinoma cells into femoral region. All parameters were assessed after 28 days post-inoculation or one-week thereafter. ZF treatment resulted in significant reduction of tumor weights with marked decrease in IL-6 and VEGF levels in serum, and tumor Ag II and CEA contents. Additionally, the administration of ZF downregulated the tumor gene expression of cyclin-D, ACE-1, and Bcl2 and upregulated the proapoptotic gene, BAX. Moreover, ZF increased CBS gene expression, which is a major contributor to cellular H2S production. In addition, ZF was able to reduce the protein expression of PI3K, pAKT, pGSK-3ß, and NFκB. Our study has provided novel insights into the possible mechanisms by which ZF may produce its tumor defeating properties. These intersecting trajectories involve the interference between PI3K/Akt and CBS signaling pathways

Cerebral infarction complicated with multiple arterial thrombosis caused by cystathionine beta-synthase gene mutation in youth: a case report / 中华神经科杂志

Hyperhomocysteinemia (HHcy) is one of the independent risk factors for youth cerebral infarction. Gene mutation of key enzymes in homocysteine metabolism is the main cause of HHcy. Few cases of cystathionine beta-synthase (CBS) compound heterozygous mutation complicated with pulmonary embolism and lower extremity artery embolism have been reported. This article reported a young cerebral infarction patient complicated with pulmonary embolism and lower extremity artery embolism, who was subsequently detected with significantly elevated blood Hcy, and finally etiologically diagnosed with CBS 833 T>C/1082C>T compound heterozygous mutation. With the treatment of folic acid, methyl cobalt amine, vitamin B 6 and anticoagulant, the blood Hcy has been gradually declined, and no new thrombotic events occurred during the follow-up period of a year.

The Role of CBS in Injury Time Estimation after Brain Contusion / 法医学杂志

OBJECTIVES@#To observe the changes of cystathionine β-synthase （CBS） expression in the cerebral cortex after brain contusion at different times.@*METHODS@#An experimental model of traumatic brain injury （TBI） in mice was established by an improved weight-drop device. Then Western blotting and immunohistochemical examination were used to detect the CBS expression in cerebral cortex around injury at different time points （1 h, 6 h, 12 h, 1 d, 2 d, 3 d, 7 d）.@*RESULTS@#The results of Western blotting revealed that the expression level of CBS was down-regulated and reached its lowest level at the 3rd days after injury, and then restored to normal level after 7 days. The results of immunohistochemistry showed that CBS was present in the normal brain cortex. CBS expression gradually decreased at the 3rd days after injury, and then restored to normal level after 7 days.@*CONCLUSIONS@#CBS has the potential to be a reference index for time estimation after brain contusion in forensic practice.

The Role of CBS in Injury Time Estimation after Brain Contusion / 法医学杂志

Objective T o observe the changes of cystathionine β-synthase (C B S ) expression in the cere-bral cortex after brain contusion at different tim es. Methods A n experim ental m odel of traum atic brain injury (T B I) in m ice w as established by an im proved w eight-drop device. T hen W estern blotting and im m unohistochem ical exam ination w ere used to detect the C B S expression in cerebral cortex around in-jury at different tim e points (1 h, 6 h, 12 h, 1 d, 2 d, 3 d, 7 d). Results T he results of W estern blotting revealed that the expression level of C B S w as dow n-regulated and reached its low est level at the 3rd days after injury, and then restored to norm al level after 7 days. T he results of im m unohistochem istry show ed that C B S w as present in the norm al brain cortex. C B S expression gradually decreased at the 3rd days after injury, and then restored to norm al level after 7 days. Conclusion C B S has the potential to be a reference index for tim e estim ation after brain contusion in forensic practice.

Polymorphisms in methylenetetrahydrofolate reductase and cystathionine beta-synthase in oral cancer - a case-control study in southeastern Brazilians / Polimorfismos em metilenotetrahidrofolato redutase, cistationina beta-sintase no câncer de boca - um estudo de caso-controle no Sudeste brasileiro

ABSTRACT INTRODUCTION: Oral squamous cell carcinoma (OSCC) is a serious public health problem, due to its high mortality rate and worldwide rising incidence. OSCC susceptibility is mediated by interactions between genetic and environmental factors. Studies suggest that genetic variants encoding enzymes involved in folate metabolism may modulate OSCC risk by altering DNA synthesis/repair and methylation process. OBJECTIVE: The goals of this study were to evaluate the association of three genotypic polymorphism (MTHFR C677T, MTHFR A1298C and CBS 844ins68) and oral cancer risk in southeastern Brazilians and evaluate the interactions between polymorphisms and clinical histopathological parameters. METHODS: This case-control study included 101 cases and 102 controls in the state of Espírito Santo, Brazil. MTHFR genotyping was done by PCR-RFLP (polymerase chain reaction - restriction fragment length polymorphism) and CBS genotyping by PCR (polymerase chain reaction) analysis. RESULTS: MTHFR C677T polymorphism was associated with lymph node involvement. Genotype CT + TT acted as a protective factor. MTHFR A1298C AC + CC genotype was associated with tumor differentiation, and possibly with a better prognosis. In risk analysis, no correlation was observed between genotypes and OSCC. CONCLUSION: We concluded that MTHFR C677T, MTHFR A1298C and CBS 844ins68 polymorphisms were not associated with OSCC risk in southeastern Brazilians; however, we suggest a prognosis effect associated with MTHFR C677T and A1298C polymorphisms in OSCC.

Resumo Introdução: O carcinoma espinocelular oral (CECO) trata-se de um importante problema de saúde pública, devido à elevada taxa de mortalidade e incidência crescente em todo o mundo. A susceptibilidade ao CECO é mediada por interações entre fatores genéticos e ambientais. Estudos sugerem que as variantes genéticas que codificam as enzimas envolvidas no metabolismo do folato podem modular o risco de CECO, alterando a síntese/reparação do DNA e o processo de metilação. Objetivo: Os objetivos deste estudo foram avaliar a associação de três polimorfismos genotípicos (MTHFR C677T, MTHFR A1298C e CBS 844ins68) e o risco de câncer oral em brasileiros da região Sudeste, e avaliar as interações entre polimorfismos e parâmetros clínico-histopatológicos. Método: Este estudo de caso-controle incluiu 101 casos e 102 controles no estado do Espírito Santo, Brasil. A genotipagem do polimorfismo MTHFR foi realizada por PCR-RFLP (Reação de Polimerase em Cadeia - Polimorfismo no Comprimento de Fragmento de Restrição) e a do CBS por análise da PCR (Reação de Polimerase em Cadeia). Resultados: O polimorfismo MTHFR C677T foi associado ao envolvimento de gânglios linfáticos. O genótipo CT + TT atuou como um fator protetor. O genótipo MTHFR A1298C AC + CC foi associado à diferenciação do tumor e, possivelmente, a um prognóstico melhor. Na análise de risco, a correlação entre os genótipos e o CECO não foi observada. Conclusão: Concluímos que os polimorfismos MTHFR C677T, MTHFR A1298C e CBS 844ins68 não estão associados ao risco de CECO nos brasileiros da região Sudeste; no entanto, sugerimos um efeito prognóstico associado aos polimorfismos MTHFR C677T e A1298C em CECO.

Expressions of hydrogen sulfide and its synthases in bladder cancer cell lines and their effects / 吉林大学学报(医学版)

Objective:To explore the expressions of endogenous hydrogen sulfide (H2 S)and its synthases cystathionine beta synthase (CBS)and cystathionine gamma lyase (CSE)in the cell lines of normal bladder and bladder cancer,and to clarify their mechanism in the development of bladder cancer.Methods:The bladder cancer cell lines (5637,T24,UM-UC-3,EJ)and human bladder epithelial cell line SV-HUC-1 were selected.The expressions of CBS and CSE in bladder cancer and normal cell lines were analyzed by Western blotting assay and the productivities of H2 S in cell lines were detected by sensitive sulphur electrode assay.The EJ cells were selected based on the previous experimental results and divided into groups as follows:① 10 μmol· L-1 NaHS group, 50 μmol·L-1 NaHS group,100 μmol·L-1 NaHS group and control group.After drug treatment,the cell survival rate was measured by MTT assay at 24 and 48 h.② 5 μg·L-1 cisplatin group,cisplatin (5 μg·L-1 )+ NaHS (100 μmol·L-1 )group and control group.After medicine treatment,the cell survival rate was measured by MTT assay and the cell apoptotic rate was detected by flow cytometry at 48 h. Results:Compared with the normal bladder cells (SV-HUC-1),the expression levels of CBS and CSE and the productivity of H2 S in the bladder cancer cell lines (5637,T24,UM-UC-3 and EJ)were increased obviously (P <0.05 or P <0.01).Compared with control group,exogenous H2 S promoted the cell proliferation of EJ cells.The cell survival rates were increased with the increase of drug dose (P <0.05),which showed a dose-dependent effect.The cell survival rates were increased with the prolongation of time (P <0.05),which showed a time-dependent effect.After medicine treatment,compared with cisplatin group,the cell viability in cisplatin+NaHS group was increased (P <0.05)and the apoptotic rate was decreased (P <0.05).Conclusion:Endogenous H2 S and its synthases CBS and CSE have an increased expression level in bladder cancer cell lines compared with the normal bladder cells.H2 S can enhance the proliferation of bladder cancer cells and decrease the apoptosis induced by cisplatin.

Effects of exogenous H2S on H2S concentration and cystathionine β-synthase expression in hippocampus in a rat model of vascular dementia / 国际脑血管病杂志

Objective To investigate the effect of exogenous hydrogen sulfide (H 2 S) on H2 S concentration and cystathionine β-synthase (CBS) expression in hippocampus in a rat model of vascular dementia (VaD). Methods A rat model of VaD was induced by using the modified four -vessel occlusion. The rats were divided into sham operation, model, low -dose and high-dose NaHS groups using the random number table method. They were further redivided into one day, seven -day, and 30-day subgroups according to the time after modeling. After modeling respectively, NaHS 30 μmol/kg and 100 μmol/kg were injected intraperitoneally every day in the low -dose and high-dose NaHS groups. The normal saline was injected intraperitoneally every day in the sham operation group and the VaD model group. Morris water maze test was used to evaluate the learning and memory ability of the rats. The expression of CBS in hippocampus was detected by real-time fluorescent polymerase chain reaction. Western Blotting was used to detect expression of CBS protein in hippocampus. Results Morris water maze test showed that the escape latencies of the model group, low -dose and high-dose NaHS groups were prolonged significantly compared with the sham operation group (P <0.05); the times of crossing the platform were decreased significantly compared with the model group (P <0.05); and the escape latencies were shortened significantly in the low -dose and high-dose NaHS groups compared with the model group ( P <0.05). The H2 S content in hippocampus was decreased significantly in the model group, low -dose and high-dose NaHS groups compared with the sham operation group, but the low -dose and high-dose NaHS group was significantly higher than that in the model group (all P <0.05). The expression of CBS mRNA and protein in the model, low -dose and high-dose NaHS groups was significantly lower than that of the sham operation group (all P < 0.05), and there was no significant difference between the low -dose and high-dose NaHS groups and the model group. Conclusions Exogenous H2 S may improve the learning and memory ability of the VaD rats. It may be associated with the increased H2 S content in hippocampus. However, it has no effect on CBS expression.

The relationship between expression of endogenous cystathionine beta-synthase and hydrogen sulfide in rectal mucosa tissue and the activeness of ulcerative colitis / 中国医师进修杂志

Objective To study the expression of endogenous cystathionine-β-synthase (CBS) and hydrogen sulfide in patients with ulcerative colitis (UC), and explore their possible role in the pathogenesis of UC. Methods Thirty patients with active period UC (active period UC group), 30 patients with remission period UC (remission period UC group) and 30 healthy controls (control group) were selected, and SABC method was used to observe the localization of CBS in rectal mucosal tissues. The optical density value of CBS was analyzed with image analysis systems. The relative expression of CBS mRNA in the rectal mucosa tissue was detected by reverse transcriptase-polymerase chain reaction (RT-PCR), and the expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was detected as an internal reference. The expression level of mRNA was detected by semi quantitative analysis with gel imaging analysis system, and the results were expressed as the ratio of the target bands to the GAPDH absorbance. The serum level of hydrogen sulfide was detected. Results The serum level of hydrogen sulfide, optical density value of CBS and relative expression of CBS mRNA of rectal mucosal tissues in active period UC group were significantly higher than those in remission period UC group and control group: (90.13 ± 3.12) μmol/L vs. (50.34 ± 2.34) and (48.13 ± 2.75) μmol/L, 0.433 ± 0.037 vs. 0.295 ± 0.064 and 0.214 ± 0.026, 1.532 ± 0.134 vs. 1.031 ± 0.107 and 0.986 ± 0.067, and there were statistical differences (P 0.05). Conclusions The abnomal expression of CBS and hydrogen sulfide in aactive period UC may play an important role in the pathogenesis of UC.

Association between cystathionineβsynthase gene T833C polymorphism and essential hypertension in Xinjiang Kazakh population and Han population / 天津医药

Objective To investigate the association between plasma homocysteine (Hcy) levels and cystathionineβsynthase (CBS) T833C gene polymorphism with essential hypertension in Xinjiang Kazakh and Han populations. Methods A total of 239 Kazak patients with hypertension (Kazak EH group), 206 Kazak people with normal blood pressure (Kazak con?trol group), 256 Han patients with hypertension (Han EH group) and 206 Han people with normal blood pressure (Han con?trol group) were selected for the study. Amplification refractory mutation system(ARMS) was used to analyze the polymor?phism of CBS gene T833C,TT,TC and CC genotypes and the various sites of T,C allele frequencies in four groups. In the meantime, the Hcy level and related biochemical indices were detected using automatic biochemical analyzer. Results The plasma Hcy levels were significantly higher in Kazak EH group and Han EH group than those of Kazak control group and Han control group (P0.05).Conclusion The Cystathionineβsynthase gene of T833C polymorphism may be associated with essential hypertension in Kazak people in Xinjiang, but no such association in Han population in Xinji?ang. The mechanism may be related to the altered metabolism of Hcy induced by CBS mutation.

Expression,purification and identification of I278T-mutant human cystathionine βsynthase / 国际检验医学杂志

Objective To investigate the expression and purification I278T-mutant human cystathionineβsynthase(CBS) in E . coli .Methods Site-directed mutagenesis by overlap extension using the polymerase chain reaction (PCR) was employed to construct mutant plasmids pGEX4T-1-CBS(I278T) ,which was induced and expressed in a medium containing 3% ethanol ,purified by affinity chromatography to obtain mutated CBS (I278T) protein .The activity ,UV-visible absorption spectroscopy ,protein particle size and Zeta potential of the purified protein were measured .Results Plasmid pGEX4T-1-CBS(I278T) was successfully constructed .The yield ,the specific activity and activity recovery of purified mutant CBS (I278T ) protein were 2 .3 mg/L ,21 .4 U/mg and 22 .6% .S-adenosylmethionine(AdoMet) with final concentration of 1 mmol/L showed no activation toward mutant CBS (I278T) protein .Ac-cording to UV-visible absorption spectroscopy analysis ,purified mutant CBS(I278T) had characteristic absorption peaks at 429 nm and 550 nm for heme-binding proteins .Protein average particle size was 7 .5 -10 .1 nm ,mainly in the form of tetramers ,and Zeta potential was - 16 .3 mV .Conclusion The methods of expression ,purification and identification of I278T-mutant human cystathionineβsynthase in E .coli were successfully established .

Effects of fluorosis on levels of cystathionine beta-synthase and hydrogen sulfide in rats / 中华地方病学杂志

Objective To observe the effects of fluorosis on the levels of endogenous cystathionine beta-synthase (CBS) and hydrogen sulfide (H2S) in rats.Methods According to body weight,forty-eight Sprague-Dawley rats (body weight 105-180 g) were divided into three groups by a random number table(16 rats in each group,half male).Fluorine contents of the feed in control group,low-fluoride group and high-fluoride group were 9.80,15.40 and 23.80 mg/kg.After 6 months of fluorine exposure,the fluorine contents of urine and bone were determined by the method of fluorine ion-selective electrode ; H2S levels in serum and brain and the activity of CBS in brain were detected by methylene blue; and protein expression of CBS was detected by Western blotting.Results Compared with control group,dental fluorosis was found in rats of low-fluoride and high-fluoride groups.The differences of fluorine contents of urine and femur were statistically significant between groups(F =65.16,67.93,all P ＜ 0.05).The urinary and femoral fluorine in low-fluoride groups [(5.25 ± 0.45)mg/L,(1 196.54 ± 72.78)mg/kg] and high-fluoride groups[(13.17 ± 0.98)mg/L,(2 656.61 ± 170.12)mg/kg] were higher than those of control groups [(3.64 ± 0.20)mg/L,(870.71 ± 71.51)mg/kg,all P ＜ 0.05],and the increases were in a dose-dependent fashion(all P ＜ 0.01).The differences of H2S contents in serum and brain were statistically significant(F =4.83,1 456.13,all P ＜ 0.05).The H2S content in serum was higher in high-fluoride group [(17.64 ± 2.38) μ mol/L] than that of the control group [(10.29 ± 0.74) μ mol/L,P ＜ 0.01].The H2S contents in brain were higher in the low-fluoride [(364.74 ± 2.06)μmol/L] and high-fluoride groups [(513.43 ± 4.18) μmol/L] than those of the control group[(314.94 ± 0.72)μmol/L,all P ＜ 0.01],and the increase was in a dose-dependent fashion (P ＜ 0.01).The difference of CBS activity was statistically significant between groups (F =760.63,P ＜ 0.01).The CBS activities were lower in low-fluoride [(438.90 ± 2.83) mmol· kg-1· min-1] and high-fluoride groups [(529.83 ± 2.37)mmol· kg-1· min-1] than those of the control group [(596.33 ± 2.75) mmol · kg-1· min-1,all P ＜ 0.01],whereas the protein expression of CBS in brain in high-fluoride group (1.49 ± 0.08) was higher than that of the control group (1.19 ± 0.06,P ＜ 0.05).Conclusion Chronic fluorosis can affect the levels of endogenous CBS and H2,S,and the increases are in a dose-dependent fashion in addition to CBS activity.

Effect of sevoflurane pretreatment on expression of cystathionine β-synthase and heme oxygenase-1 during myocardial ischemia-reperfusion injury in rats / 中华麻醉学杂志

Objective To investigate the effect of sevoflurane pretreatment on the expression of cystathionine β-synthase(CBS)and heme oxygerase-1(HO-1)during myocardial ischemia-reperfusion(I/R)injury in rats.Methods Thirty adult male SD rats,weighing 180-220 g,were randomly divided into 3 groups(n =10each):sham operation group(group S),I/R group and sevoflurane group(group Sev).Myocardial I/R injury was produced by ligation of the left anterior descending branch of coronary artery for 30 min followed by 2 h reperfusioo.In group Sev,sevoflurane was inhaled before ischemia,the end-tidal concentration was 1.5％-1.7％ and myocardial I/R was produced 60 min later.The rats were sacrificed at 2 h of reperfusion and myocardial tissues were taken for determination of the contents of MDA,GSH,H2S and CO,SOD activity and expression of CBS mRNA and HO-I mRNA.The ultrastructure of myocardium was examined with electron microscope.Results Compared with group S,the contents of MDA,H2S and CO were significantly increased,the expression of CBS mRNA and HO-1 mRNA was up-regulated,and SOD activity and GSH content were significantly decreased in groups I/R and Sev(P ＜ 0.05).Compared with group I/R,the contents of MDA,H2S and CO were significanfly decreased,the expression of CBS mRNA and HO-1 mRNA was down-regulated,SOD activity and GSH content were significantly increased in group Sev.The damage to mitochondrial structure induced by I/R was mitigated by pretreatment with sevoflurane.Conclusion The mechanism by which sevoflurane pretreatment reduces myocardial I/R injury is related to down-regulation of the expression of CBS and HO-1 and decrease in the activities in rats.

Effect of aminooxoacetic acid on focal cerebral ischemia injury in rats / 中华麻醉学杂志

ObjectiveTo evaluate the effect of aminooxyacetic acid on focal cerebral ischemia injury in rats.MethodsEighty healthy male SD rats aged 2.5 month weighing 250-280 g were randomly divided into five groups( n ＝ 16 each):sham operation group(group S),cerebral ischemia group(group Ⅰ),aminooxoacetic acid low,medium and high dose groups(groups AL,AM and AH ).Focal cerebral ischemia was induced by occlusion of middle cerebral artery using a nylon thread with rounding tip which was inserted into right internal carotid artery in groups I,AL,AM and AH.Intraperitoneal amincoxoacetic acid 25,50 and 100 μmol/kg were administered at 3 h of ischemia in groups AL,AM and AH respectively,while equal volume of normal saline 1 ml/kg were injected in groups S and I.Neurological function was assessed and scored (0＝ no deficit,4＝ unable to move,unconscious) in 8 rats at 21 h after aminooxyacetic acid administration in each group.The animals were then sacrificed and the brains were removed for determination of the cystathionine beta-synthase (CBS) activities in cortex,hippocampus and striatum corpora.The other eight rats of each group were sacrificed at 21 h after amincoxoacetic acid administration for determination of the cerebral infarct volume.ResultsCompared with group S,the neurological deficit scores and the CBS activities in cortex and hippocampus were significantly increased,the infarct volumes were significantly enlarged in group Ⅰ ( P < 0.05).Compared with group Ⅰ,the neurological deficit scores and the CBS activities in cortex and hippocampus were significantly decreased,the cerebral infarct volumes were significantly reduced in groups AM and AH (P < 0.05).There was no significant difference in the above-mentioned variables between groups AL and Ⅰ.ConclusionAminooxoacetic acid can reduce focal cerebral ischemia injury by decreasing CBS activity and reducing H2 S production in rats.

Niveles de homocisteína y polimorfismos de los genes de la MTHFR y la CBS en pacientes colombianos con trombosis venosa superficial y profunda / Homocysteine levels and polymorphisms of MTHFR and CBS genes in Colombian patients with superficial and deep venous thrombosis

Introducción. Se produce trombosis cuando en el sistema hemostático se desequilibran los mecanismos procoagulantes, anticoagulantes y fibrinolíticos, y se forman coágulos dentro de los vasos sanguíneos. Los factores de riesgo para el desarrollo de esta enfermedad pueden ser adquiridos o genéticos, polimorfismos o mutaciones en genes que conducen a hiperhomocisteinemia o que están comprometidos en las vías de coagulación. Objetivos. Analizar, en una población colombiana con diagnóstico de trombosis venosa el perfil lipídico, los niveles de glucosa y homocisteína, y calcular las frecuencias alélicas y genotípicas de los polimorfismos c.677C>T del gen de la metilen-tetra-hidrofolato reductasa (MTHFR) y c. 699C>T, c.1080 C>T, c.844ins68 del gen de la cistationina betasintasa (CBS).Materiales y métodos. Se estudiaron 33 pacientes con sus respectivos controles. Las pruebas bioquímicas se realizaron por métodos colorimétricos y de inmunoensayo. Se utilizó la técnica de fragmentos de longitud polimórfica para la identificación de los polimorfismos mencionados. El estudio de asociación se hizo mediante la prueba de de ji al cuadrado. Resultados. Se confirmó el papel de algunos factores de riesgo ya establecidos para el desarrollo de enfermedad trombótica venosa y se encontró un efecto protector del polimorfismo CBS c.699C>T para el riesgo de hipercolesterolemia con diferencia estadísticamente significativa en el grupo de los casos al compararlo con los controles. Por otra parte, se encontró una tendencia estadística que podría indicar un efecto protector del polimorfismo 844ins68 para el desarrollo de enfermedad trombótica venosa. Conclusiones. No se encontraron diferencias estadísticamente significativas en los niveles de homocisteína entre el grupo de casos y de controles. Sin embargo, la variabilidad en las concentraciones plasmáticas fue mayor en los casos.

Introduction. Thrombosis develops when the hemostatic system is incorrectly activated due to the unbalance between procoagulant, anticoagulant and fibrinolytic mechanisms allowing the formation of a clot within a blood vessel. The risk factors of this pathology can be acquired or can be genetic. Objectives. To analyze in a Colombian population with diagnosis of venous thrombosis, lipid profile, glucose and homocystein levels, to calculate the alleles and genotypic frequencies of polymorphisms c.699 C>T, c.1080 C>T, c.844ins68 of the cystathionine ß synthase and the c.677 C>T of the methylenetetrahydrofolate reductase (MTHFR) genes. Materials and methods. Thirty three patients and their controls were studied. The biochemical test was carried out by colorimetric methods and immunoassay. In this survey we used the restriction fragments longitude polymorphism (RLFP) technique to identify the polymorphisms mentioned. The association study was performed through the chi square test. Results. We confirmed that gene alterations increase risk for pathology; we found statistically significant differences in the group with hypercholesterolemia in presence of the polymorphism c.699 C>T in the CBS gene, showing a protective effect in the individuals carrying this genetic variation. Likewise, we found a statistical trend for an eventual protective effect of the CBS c.844ins68 polymorphism to venous thrombotic disease. Conclusions. There were not any statistically significant differences in homocystein levels between cases and controls; nevertheless, the variability in the plasma concentrations was greater in the group of cases.

Gaseous signal molecule hydrogen sulfide and penile erection / 第二军医大学学报

Endogenous hydrogen sulfide is the third gaseous signal molecule after nitric oxide and carbon monoxide; it is synthesized through cystathionine- β-synthase and cystathionine-γ-cleavage pathway. Hydrogen sulfide can directly react with the KATP channel, reduce extracellular Ca 2+ influx, and work together with testosterone, nitric oxide, carbon monoxide, relaxing corpus cavernosum smooth muscle and improving diastolic function of blood vessels. It can also inhibit vascular smooth muscle cell proliferation, promote apoptosis, and improve vascular remodeling. Therefore, in-depth study of H2 S in the regulating penile erection can provide a new way for the treatment of erectile dysfunction.

Volatile Sulfur Compounds as a Predictor for Esophagogastroduodenal Mucosal Injury

BACKGROUND/AIMS: Halitosis is a symptom that bothers patients more socially than medically and its pathogenic mechanisms are unclear and treatment armamenterium is limited. Clinicians generally ignored active interventions. Since halitosis is closely associated with volatile sulfur compounds (VSCs), we used a Halimeter and gas chromatography to measure VSCs in patients with Helicobacter-pylori (H. pylori)-associated gastric diseases. METHODS: We categorized 72 patients with H. pylori infection into two groups based on their endoscopic findings: a non-erosive mucosal group (NE, n=24) and an erosive mucosal group (E, n=48). Halitosis was objectively assessed by applying either a Halimeter to breath air or gas chromatography to gastric juice. Simultaneously, the expression of VSC-generating enzyme was measured with reverse-transcriptase PCR using mRNA isolated from biopsy tissues. RESULTS: The levels of VSCs in exhaled breaths or aspirated gastric juices differed significantly between the NE and E groups (p<0.00001), suggesting that VSCs might reflect eroded epithelial damage induced by H. pylori infection. The expressions of cystathionine beta-synthase (CBS) and cystathionine gamma-lyase (CSE) were broadly consistent with the degree of mucosal injury. CONCLUSIONS: Erosive changes in esophagogastroduodenal mucosa were strongly correlated with increased VSC levels, suggesting that halitosis might result from H. pylori-associated erosive lesions.

Recombinant adeno-associated virus mediated gene transfer in a mouse model for homocystinuria

Homocystinuria is a metabolic disorder caused by a deficiency of cystathionine b-synthase (CBS). The major clinical symptoms of this disease are mental retardation, lens dislocation, vascular disease with life-threatening thromboembolisms, and skeletal deformities. The major treatments for CBS deficiency include pharmacologic doses of pyridoxine or dietary restriction of methionine. There is currently no effective long-term treatment to lower the elevated plasma levels of homocysteine. However, gene therapy could be an effective novel approach for the treatment of homocystinuria. A recombinant adeno- associated virus vector carrying human CBS cDNA (rAAV-hCBS) was constructed and administered to CBS-/- mice by intramuscular (IM) and intraperitoneal (IP) injections. Serum homocysteine concentrations significantly decreased in treated mice compared with age-matched controls two weeks after treatment. The treated CBS-/- mice had life spans 3-7 days longer compared with untreated CBS-/- mice. In CBS-/- mice treated with rAAV-hCBS via IP injection, the vector was detected in all organs examined including liver, spleen, and kidney, and CBS gene expression was observed by immunohistochemical staining in the liver. These results indicate the efficacy of gene delivery and demonstrate the possibility of gene therapy mediated by AAV gene transfer in this mouse model of homocystinuria.

Association between MTHFRC677T,CBS844ins68 polymorphism and young ischemic stroke / 吉林大学学报(医学版)

Objective To investigate the genetic association of MTHFRC677T between CBS844ins68 polymorphism and young ischemic stroke.Methods The polymorphism of MTHFRC677T and CBS844ins68 in 98 stroke patients and 116 control subjects were examined and analyzed by using PCR-RFLP.Results The frequencies of two alleles were as follows: T allele 54.09%,C allele 45.91% in patients and T 37.93%,C 62.07% in controls(?2=11.18,P

Alteration of hydrogen sulfide/cystathionine-?-synthase system in rats with recurrent febrile seizures / 北京大学学报(医学版)

Objective: To study the alteration of hydrogen sulfide (H_2S)/ cystathionine-?-synthase (CBS) system during recurrent febrile seizures (FS) in the hippocampus of developing rats. Methods: The rats were randomly divided into control group (n=8) and hyperthermia-treated group (n=22). Which was subdivided into FS group (n=8) and H group(no seizure occurred, n=9) according to whether seizures occurred. The plasma level of H_2S was detected by the spectrophotometer. The expression levels of CBS gene and protein were examined by in situ hybridization and immunohistochemistry respectively. Results: The plasma levels of H_2S were increased significantly in FS group compared with those of control group or H group. The expression levels of CBS gene and protein were enhanced in FS group compared with those of control group or H group. Conclusion: The expression levels of H_2S/ CBS system were up-regulated during recurrent FS.

Plasma Homocysteine Concentration and Genotype Variation of Enzymes as Risk Factors in Patients with Coronary Artery Disease

BACKGROUND AND OBJECTIVES: Increased plasma homocysteine(tHcy) has been implicated as an independent risk factor for coronary artery diseas(CAD), but the relationship has not been firmly established. Present study aimed to determine the difference of plasma homocysteine between patients with CAD and normal control, and to identify the relation between plasma homocysteine and genotype variation of its metabolic enzymes, and serological characteristics. METHODS: Plasma homocysteine, fasting and post-methionin loading, folate and vitamin B12 were measured among 149 patients and 80 control subjects. Both group consisted of those younger than 65 years. Frequencies of prevalent mutations of enzymes involved in homocysteine metabolism, cytosine to thymidine transition (C(677)T) of methylentetrahydrofolate reductase (MTHFR) was determined by polymerase chain reaction (PCR) in 85 patients and 47 control. RESULT: There was no significant difference in homocysteine level between patients and control group (fasting tHcy; 10.4 +/- 3.6 vs 11.4 +/- 8.4 ng/ml, post-methionine loading tHcy; 18.8 +/- 4.9 vs 17.2 +/- 9.5 ng/ml, p> 0.05 respectively). Genotype frequency of MTHFR C(677)T was similar between two groups. Plasma homocysteine level did not appear to vary with genotypes of MTHFR both in patients and control subjects. Multiple linear regression analysis identified smoking as the most significant factor affecting plasma homocysteine level, followed by age, MTHFR genotype, obesity, and folate level. CONCLUSION: Homocysteine concentration was not different between controls and patients with CAD. Significant variation of homocysteine level according to genetypic polymorphism of metabolism enzymes was not observed. On multiple linear regression, several factors were identified to be related to homocysteine level, including MTHFR genotype. Further study is warranted to clarify the significance of homocysteine in the development of CAD.