Objective To investigate the effect of carbon monoxide(CO) on hydrogen sulfide(H_2S)/cystathionine-?-lyase system(CSE) in vascular smooth muscle cells(VSMC) of spontaneous hypertensive rats(SHR).Methods The SHR VSMC were divided into 2 groups:experimental group(hemin was added to the culture media at a concentration of 10 ?mol/L)and control group (dimethyl sulfoxide was added to the culture media at an equal volume).The content of H_2S in the cultrue media was measured by sulfide electrode method,and the CSEmRNA level was assayed by competitive reversed transcription-polymerase chain reaction(RT-PCR).Results Compared with control group,the content of H_2S in the culture media of hemin-treated SMCs was significantly lower[(16.03? 2.14) ?mol/L vs (13.31?1.74)?mol/L],and the CSEmRNA level in hemin-treated SMCs was decreased markedly(0.17?0.04 vs 0.13?0.03).Conclusion CO can down-regulate the H_2S/CSE system in SMC of SHR.

Objective To investigate the effect of nitric oxide on hydrogen sulfide/ cystathionine-?-lyase(CSE) system in the vascular smooth muscle cells of spontaneously hypertensive rats(SHR).Methods The SHR aortic smooth muscle cells(SMCs) were divided into 2 groups: sodium nitroprusside(SNP) group and control group.The content of hydrogen sulfide in the culture media was measured by sulfide electrode method,and the CSE mRNA level was assayed by competitive reversed transcription-polymerase chain reaction((RT-PCR)).Results Compared with control group,the content of hydrogen sulfide in the culture media of SNP group was significantly higher [(16.16?3.71) ?mol/L vs(22.71?4.84) ?mol/L],and the CSE mRNA level in SMCs in SNP group was lower than that of control group.Conclusion Nitric oxide exerted complicated effect on the hydrogen sulfide/CSE system in the SHR smooth muscle cells.J Appl Clin Pediatr,2006,21(3):140-141

OBJECTIVETo explore the differences of electroacupuncture (EA) on onset time and symptom improvement for treatment of different types of functional constipation.

METHODSThirty-eight cases of constipation were selected, including 9 cases of constipation-predominant irritable bowel syndrome (IBS-C), 10 cases of slow transit constipation (STC), 10 cases of pelvic floor dyssynergia (PFD) and 9 cases of inadequate defecatory propulsion (IDP). The electroacupuncture was applied at Tianshu (ST 25), Fujie (SP 14), Shenshu (BL 23), Dachangshu (BL 25) and so on in abdominal and lumbosacral area, 5 times per week, 10 times as a treatment course. The onset time, score of clinical symptoms of constipation and improvement of every symptom in each group were compared.

RESULTS(1) The onset time was (1.78 +/- 0.83) days in IBS-C type, (3.11 +/- 1.90) days in IDP type, (4.10 +/- 1.85) days in STC type and (4.30 +/- 2.00) days in PFD type, indicating statistical differences between IBS-C type and STC type, IBS-C type and PFD type (both P < 0.05). (2) Compared before the treatment, the total scores of symptoms on the onset day in each group were all improved (P < 0.05, P < 0.01), and score of IBS-C type was superior to the rest 3 types (P < 0.05, P < 0.01). (3) EA improved desire to defecate or frequency of defecation in each type (P < 0.05, P < 0.01), in which both were improved in STC type and PFD type, and the improvement of defecation frequency was more significant in STC type (P < 0.01). EA relieved unsmooth defecation or pendant-expansion feeling in each type (all P < 0.05), in which both were improved in IBS-C type (both P < 0.05). EA relieved abdominal distension and pain in IBS-C type, STC type and PFD type (all P < 0.05), while its effects were not obvious on defecation difficulty, defecation time and defecation texture (all P > 0.05).

CONCLUSIONThe electroacupuncture for treatment of 4 types of constipation is characterized by rapid onset; the improved symptoms are not identical in the electroacupuncture treatment plan for each type of defecation; the main improvement of symptoms are lied on desire to defecate and frequency of defecation, unsmooth defecation or pendant-expansion feeling and abdominal distension and pain. Meanwhile the improvements of defecation texture, defecation difficulty and defecation time were not signi-ficant.

Adult ; Aged ; Constipation ; etiology ; physiopathology ; therapy ; Defecation ; Electroacupuncture ; Female ; Humans ; Irritable Bowel Syndrome ; complications ; Male ; Middle Aged ; Pelvic Floor Disorders ; complications ; Treatment Outcome ; Young Adult

Objective To investigate the effects of maternal long-term exposure to low-dose trichlorfon on the serum paraoxonase (PON) activity of pregnant mice and development of embryos. Methods Female ICR mice (n=120) were randomly divided into control group and trichlorfon groups of different doses,and were managed by intragastric injection with trichlorfon of 0,2,10 and 50mg/kg,respectively. All the mice were managed once a day for a consecutive of 27 days,and were subjected to mating. The pregnant mice were continued to be managed with trichlorfon for 3 days,and were sacrificed on day 3 of gestation. The serum PON and acetylcholinesterase (AchE) activities were detected,and the development of embryos was evaluated. Results The serum PON activity of 2,10 and 50mg/kg trichlorfon group were (14.15?1.22),(12.78?1.80) and (10.45?1.95)IU/mL,respectively,and that of 50mg/kg trichlorfon group was significantly lower than that of control group [(13.37?2.31)IU/mL] (P0.05),while the the percentage of abnormal embryos of 50mg/kg trichlorfon group had an increased tendency. Conclusion Long-term exposure to low-dose trichlorfon can inhibit serum PON and AchE activity in pregnant mice without obvious effect on the development of embryos.

Objective To explore the expression and distribution of transient receptor potential cation channel 6(TRPC6)in normal human,mice,rats' renal tissue and the mouse podocyte clone 5(MPC5)for further investigating the relationship between TRPC6 and the protei-nuria-related podocyte molecules.Methods 1.The distributions of TRPC6 in normal human,mice,rats' renal tissue and MPC5 were observed by using the immunochemistry staining.2.The mRNA expression of TRPC6 in mouse renal cortex and differentiated MPC5 was detected by using reverse transcriptase-polymerase chain reaction(RT-PCR).3.The protein expression of TRPC6 in human,mice and differentiated MPC5 was detected by using Western blotting.Results 1.In human kidney,TRPC6 showed a weak staining in glomeruli and a strong staining in renal tubules and vessels.In mice and rats' kidney,TRPC6 showed a strong staining in glomeruli and was mainly distributed along the capillary loops of glomerulus and in mesangium.The positive staining of TRPC6 was observed in MPC5,which was distributed evenly on the cell membrane in differentiated podocytes.2.The specific PCR band of TRPC6 was detected in mouse renal cortex and differentiated MPC5.3.The specific protein band of TRPC6 was detected in normal human,mice renal cortex and differentiated MPC5 with the size of 106.Conclusions The expression of TRPC6 is verified in normal human,mice and rats' kidneys,and in differentiated MPC5.These results will benefit for further exploring the relationship between TRPC6 and the proteinuria-related podocyte molecules.

Objective To explore the relationship between dynamic changes in ultra-micro-structural of pulmonary arteries and endogenous hydrogen sulfide in rats with left-right shunt.Methods Rats in shunt group were subjected to an abdominal aorta-inferior vena cava shunt to create an animal model of pulmonary artery structural remodeling. After 1 day, 3 days, 1 week, 4 weeks and 8 weeks of experiment, the ultra-micro-morphologic changes of pulmonary arteries of rats were observed under electronic microscope and H_2S concentration in serum was evaluated by modified sulfide electrode method.Results The changes of ultra-micro-structure of pulmonary arteries were progressively exacerbated, endothelial cells became swollen and large in size on 3 days, smooth muscular cells increased in size as well as the change of endothelial cells in 1 week, and they changed from contractile phenotype to synthetic phenotype in 4 weeks.Conclusions Shunt exhibited changes of ultra-micro-structure of pulmonary arteries are accompanied by the changes of endogenous H_2S. It is suggested that endogenous H_2S might play a protective role in changes of ultra-micro-structure of pulmonary artery.

Objective To evaluate the short-term efficacy and safety of sevelamer hydrochloride in treating maintenance hemodialysis (MHD) patients with hyperphosphemia.Methods A multicenter,open-labeled,self-control study was performed.Phosphate binders were discontinued during a two-week washout period.Patients with more than 1.78 mmol/L serum phosphorus after two-week washout period were eligible for the trial.The dose was adjusted every two weeks as necessary to achieve serum phosphorus control. Sevelamer hydrochloride was administered to 138 MHD patients for 10 weeks and a second two-week washout period followed.Results A total of 111 from 138 patients fulfilled the whole 14-week study. Mean serum phosphorus and calcium-phosphate products starte to decline after two-week sevelamer hydrochloride treatment. By the end of 10-week sevelamer hydrochloride treatment, mean serum level of phosphorus [(1.85±0.50) vs (2.57±0.54) mmol/L,P＜0.01],calcium-phosphate product [(4.16± 1.72) vs (5.79 ± 1.50) mmol2/L2,P＜0.01 ] and low density lipoprotein [(1.64±0.76) vs (2.31 ±0.87) mmol/L,P＜0.01] were significantly decreased,while the adjusted serum level of calcium and serum intact parathyroid hormone kept steady.Both serum phosphorus and calcium-phosphrus product increased after the second washout period, but the levels were still lower as compared to pre-treatment [(2.26±0.71) vs (2.57±0.54) mmol/L; (5.12±1.63) vs (5.79±1.50) mmol2/L2,P＜0.01].Of the 138 patients involved,214 episodes in 106 patients and 121 episodes in 89 patients were reported as adverse events and adverse drug reaction respectively. Gastrointestinal symptoms,of which most were mild or moderate,happened to 68.1％ (94/138) patients. Conclusions Sevelamer hydrochloride can control serum phosphorus and reduce the levels of calcium-phosphorus product and cholesterol.Slight gastrointestinal symptoms like constipation are common during the treatment.

AIMTo explore the time-dependent changes of endogenous hydrogen sulfide system at the early stage of pulmonary hypertension induced by high pulmonary flow in rats.

METHODSEighty male SD rats, whose weight ranged 140 - 160 g, were randomly divided into control group (n = 40) and shunt group (n = 40). Rats in shunt group were subjected to an abdominal aorta-inferior vena cava shunt to create an animal model of high pulmonary flow. After 1 d, 3 d, 1 week, 4 week and 8 weeks of experiment, systolic pulmonary artery pressure (SPAP) of each rat, the H2S of rat lung tissue and CSEmRNA of rat lung tissue were evaluated, respectively.

RESULTSSPAP increased significantly as compared with those in control group in 1 week and 8 weeks of experiment. In contrast to control group, the H2S of rat lung tissue increased significantly on 3 d and in 4 weeks, respectively. Meanwhile, in contrast to control group, relative amount of CSE mRNA of lung tissues elevated significantly on 3 d and in 4 weeks, respectively. Moreover, SPAP and the H2S of rat lung tissue, the CSE mRNA of rat lung tissue correlated negatively in 1 week, 4 weeks and 8 weeks of experiment.

CONCLUSIONAnimal model of rats with high pulmonary blood flow exhibited pulmonary hypertension. Lung tissue H2S and CSE mRNA of rats exhibited double peaks within 8 weeks. These results revealed that endogenous H2S system might be relevant with the development of pulmonary hypertension induced by high pulmonary blood flow, and probably, it played a protective role in the regulation of pulmonary hypertension, especially, at its early stage.

Animals ; Hydrogen Sulfide ; metabolism ; Hypertension, Pulmonary ; metabolism ; physiopathology ; Lung ; blood supply ; metabolism ; physiopathology ; Male ; Pulmonary Artery ; metabolism ; physiopathology ; Rats ; Rats, Sprague-Dawley

OBJECTIVE: To set up the method for analyzing HLA-B gene polymorphism with PCR-RFLP, and to gain population data among northern Chinese Hans of HLA-B's restricted fragments after NlaIII digestion, and to achieve application in forensic medicine practice. METHODS: Sample DNA was extracted by the phenol/chloroform extraction method, 943 bp-long fragments containing HLA-B exon 2 and 3 were got by PCR. The endonuclease NlaIII was applied to cut the PCR products into polymorphic fragments shorter than 943bp, then PAGE and silver staining were used to detect the digestion results, finally the digestion sites were assured by DNA sequencing. RESULTS: Along 943bp-long PCR products, 14 length-different fragments, 20 kinds of fragment combinations were got and 6 cutting site were observed after NlaIII digestion. CONCLUSION HLA-B gene was highly polymorphic among Chinese northern Hans. Even with only one endonuclease, 14 restricted fragments were got and the PIC was great. Such a HLA-B PCR-RFLP analysis will have values in forensic medicine applications.
Asian People/genetics* ; Base Sequence ; China/ethnology* ; DNA/isolation & purification* ; Exons ; Forensic Medicine/methods* ; Gene Frequency ; HLA-B Antigens/genetics* ; Humans ; Polymerase Chain Reaction/methods* ; Polymorphism, Restriction Fragment Length

OBJECTIVEPulmonary hypertension (PH) is a common complication of congenital heart defects with a left-to-right shunt characterized by high pulmonary blood flow. Pulmonary vascular structural remodeling (PVSR) is the pathological basis of PH. However, the pathophysiologic features and mechanisms responsible for PH and PVSR induced by increased pulmonary blood flow have not been fully understood. The present study was designed to explore the possible effect and mechanism of hydrogen sulfide (H(2)S) on the regulation of PVSR induced by high pulmonary flow in rats.

METHODSThirty-two male SD rats, weighing 120 - 140 g, were randomly divided into shunt group (n = 8), shunt + NaHS group (n = 8), control group (n = 8) and control + NaHS group (n = 8). Rats in shunt group and shunt + NaHS group were subjected to an abdominal aorta-inferior vena cava shunt to create an animal model of high pulmonary flow. Rats in the control and control + NaHS groups underwent the same experimental protocol as mentioned above except for the shunt procedure. Rats in the shunt + NaHS and control + NaHS groups were intraperitoneally injected with NaHS at 56 micromol/(kgxd), and rats in the shunt and control groups were injected with the same volume of physiological saline. After 11 weeks of experiment, rats were sacrificed and lung tissues were obtained. The percentage of muscularized artery (MA) was calculated. The changes in relative medial thickness (RMT) in small pulmonary arteries and median pulmonary arteries were examined. Proliferative cell nuclear antigen (PCNA), extracellular signal-regulated kinase (ERK1) and phosphorylation extracellular signal-regulated kinase (P-ERK1) protein expression were examined by Western blot, and at the same time, PCNA protein expression by pulmonary artery smooth muscle cells was observed by immunohistochemistry.

RESULTSAfter 11 weeks of shunt, compared with control group, the percentage of MA increased significantly (25.12 +/- 2.26 vs 14.42 +/- 3.41, P < 0.05), and RMT in small pulmonary arteries and median pulmonary arteries increased significantly in rats of shunt group (23.6 +/- 3.5 vs 12.6 +/- 2.1, 24.8 +/- 1.9 vs 13.5 +/- 2.2, P < 0.05 for all). PCNA protein expression in small and median pulmonary arteries increased significantly (0.49 +/- 0.04 vs 0.39 +/- 0.07, 0.46 +/- 0.08 vs 0.36 +/- 0.05, P < 0.01 for all), and the ratio of PERK/ERK1 protein expression of pulmonary arteries increased significantly (P < 0.01) in rats of shunt group compared with those of control group. After the administration of exogenous H(2)S donor, NaHS, for 11 weeks, in contrast to rats in shunt group, the percentage of MA decreased significantly (21.5 +/- 2.0 vs 25.1 +/- 2.3, P < 0.05), and RMT in small and median pulmonary arteries decreased significantly (20.2 +/- 2.8 vs 23.6 +/- 3.5, 20.8 +/- 3.1 vs 20.8 +/- 3.1, P < 0.05 for all) in rats of shunt + NaHS group. PCNA protein expression in small and median pulmonary artery smooth muscle cells decreased significantly (0.32 +/- 0.06 vs 0.49 +/- 0.04, 0.29 +/- 0.07 vs 0.46 +/- 0.08, P < 0.01 for all), and the ratio of PERK/ERK1 protein expression of pulmonary arteries decreased significantly (P < 0.01) in rats of shunt + NaHS group compared with that of shunt group.

CONCLUSIONH(2)S may play a regulatory role in pulmonary vascular structural remodeling induced by high pulmonary blood flow via mitogen-activated protein kinase (MAPK)/ERK signal transduction pathway.

Animals ; Hydrogen Sulfide ; pharmacology ; Hypertension, Pulmonary ; pathology ; physiopathology ; Lung ; pathology ; Male ; Pulmonary Artery ; drug effects ; physiopathology ; Rats ; Rats, Sprague-Dawley