AIMTo study the functional activity of K(cA) on human mesenteric artery smooth muscle cells (SMC) from essential hypertension (EH) patients.

METHODSPatch clamp technique with Inside-out mode was used for recording channel activity.

RESULTS(1) K(CA) from human mesenteric artery SMC was voltage dependent. Single channel conductance was 191.4 pS and 197.7 pS in EH group and normal group respectively. The channel activity was blocked by TEA in cytoplastic side. (2) With the increasing of Ca2+ concentration in the bath solution (fron 0 to 10(-8), 10(-7), 5 x 10(-7), 10(-6) mol/L), the K(Ca) channel open probability (Po) of two groups were increased, Po of EH group increased from 0.016 to 0.023, 0.031, 0.0153, 0.094, Po of normal group increased from 0.004 to 0.023, 0.041, 0.072, 0.184. The mean open time lengthened and the mean close time shortened. (3) Po of K(Ca) of EH group was higher than that of normal group when Ca2+ concentration was 0, but it was equal with or lower than normal group at the other Ca2+ concentrations.

CONCLUSIONK(Ca) of mesenteric artery SMC from EH group has a lower Ca2+ sensitivity than that of normal person. This might be an active factor for the onset of EH.

Calcium ; metabolism ; Case-Control Studies ; Female ; Humans ; Hypertension ; metabolism ; physiopathology ; Male ; Mesenteric Arteries ; cytology ; metabolism ; Middle Aged ; Muscle, Smooth, Vascular ; cytology ; metabolism ; physiology ; Myocytes, Smooth Muscle ; metabolism ; physiology ; Patch-Clamp Techniques ; Potassium Channels, Calcium-Activated ; physiology

OBJECTIVETo identify the relationship between angiotensin-converting enzyme (ACE) gene polymorphism and heart rate variability in cerebral stroke patients.

METHODSAn insertion/deletion(I/D) polymorphism of the ACE gene was analyzed by polymerase chain reaction in 43 normal subjects, 46 patients with ischemic cerebral stroke, and 40 patients with brain hemorrhage; their heart rate variability(HRV) parameters such as time domain and power spectral component were analyzed.

RESULTSThe frequency of DD genotype and the frequency of deletion alleles in cerebral stroke groups were significantly higher than those in control groups (P<0.01), and the measured components of HRV, including total power (TP), low frequency power (LF), high frequency power (HF), LF/HF, and choas, were higher in the patients with the ACE DD genotype when compared with those in the patients with the ACE II, ID genotypes; there was significant difference in effective rate (P<0.05).

CONCLUSIONThe above related parameters of HRV were correlated with heritability, suggesting that the cerebral stroke patients with the ACE DD genotype are at high risk of cerebrogenic cardiac autonomic nerve function disturbances.

Aged ; Female ; Genotype ; Heart Rate ; Humans ; Male ; Middle Aged ; Peptidyl-Dipeptidase A ; genetics ; Polymorphism, Genetic ; Stroke ; genetics ; physiopathology

OBJECTIVETo study regulation of Ca(2+)-activated K(+) channels (KCa) of mesenteric artery smooth muscle cell (SMC) from 21 old patients with essential hypertension (EH) by endothelin-1 (ET-1) and prostagl E(1) (PGE(1)).

METHODSMesenteric artery branch from EH was digested by enzyme. Patch clamp technique was used to pull cell-attached and inside-out patches on mesenteric artery SMC from EH and the normotensive patients respectively. The signal channel open probability (Po), open dwell-time (To) and close dwell-time (Tc), open channel number per patch were recorded. After adding Ca(2+) (10(-8) approximately 10(-6) mol/L), ET-1(2 approximately 8 x 10(9) mol/L) and PGE(1) (10, 20, 40, 100, 200, 400 nmol/L) to cytoplasm respectively. The parameters above were observed again.

RESULTSCompared to that of normotensive patients, the activities of KCa channels of patients with EH was higher. After adding Ca(2+) to cytoplasm,the Po of KCa channels in normotensive patients increased significantly. But it was few changes in EH group. KCa channels has dual reaction to ET-1 in normotensive patients. We have found no statistics difference when ET-1 present on KCa channels of EH cases. Whereas PGE(1) can affect KCa channels current and channels kinetic significantly in side-out patches. The Po of KCa channels increased. The To protracted and the Tc curtailed in EH.

CONCLUSIONSThe activities of KCa channels of patients with EH increased significantly. but the sensitive to Ca(2+) decreased. ET-1 were few effect to KCa channels. The PGE(1) can activated KCa channels of patients with EH.

Aged ; Alprostadil ; pharmacology ; Cells, Cultured ; Endothelin-1 ; pharmacology ; Female ; Humans ; Hypertension ; metabolism ; physiopathology ; In Vitro Techniques ; Male ; Mesenteric Arteries ; cytology ; Middle Aged ; Muscle, Smooth ; metabolism ; physiopathology ; Patch-Clamp Techniques ; Potassium Channels, Calcium-Activated ; drug effects ; metabolism

OBJECTIVETo determine the molecular basis of late onset ornithine transcarbamylase (OTC) deficiency in a Chinese family of Han nationality and the exon sequences of OTC gene of this patient.

METHODSPolymerase chain reaction-single strand conformation polymorphism and direct sequencing were used to identify the mutation type.

RESULTSA missense mutation E122G in the conserved residue of exon 4 was identified which is unreported before.

CONCLUSIONThe E122G mutation in human OTC gene may cause late onset OTC deficiency.

Age of Onset ; Base Sequence ; Child, Preschool ; DNA ; chemistry ; genetics ; DNA Mutational Analysis ; Family Health ; Fatal Outcome ; Female ; Humans ; Male ; Models, Molecular ; Mutation, Missense ; Ornithine Carbamoyltransferase ; chemistry ; genetics ; Ornithine Carbamoyltransferase Deficiency Disease ; enzymology ; genetics ; pathology ; Pedigree ; Polymorphism, Single-Stranded Conformational ; Protein Structure, Secondary

OBJECTIVETo develop a real-time fluorescent PCR protocol suitable for the routine screening of AZFc/DAZ microdeletions on the Y chromosome in azoospermic and oligozoospermic male infertility patients.

METHODSA set of real-time fluorescent PCR was established. Eighty-seven azoospermic and ligozoospermic patients undergoing ICSI in the IVF center and 30 azoospermic men undergoing testicular biopsy in the clinic of urology surgery were screened for AZFc/DAZ microdeletions of Y chromosome.

RESULTSEleven cases (9.4%) of AZFc/DAZ microdeletions were found in 117 cases of azoospermic and oligozoospermic patients by screening of realtime fluorescent PCR. Four cases (6.6%) were found in 61 oligozoospermic patients, and 7 cases (12.5%) were found in 56 azoospermic patients.

CONCLUSIONThe real-time fluorescent PCR protocol presented in this study is an easy and reliable method for detection of AZFc/DAZ microdeletions on the Y chromosome, which yields identical results to those of the multiplex PCR.

Chromosome Deletion ; Chromosomes, Human, Y ; Deleted in Azoospermia 1 Protein ; Fluorescence ; Humans ; Infertility, Male ; genetics ; Male ; Polymerase Chain Reaction ; methods ; RNA-Binding Proteins ; genetics

OBJECTIVETo develop a multiplex PCR protocol, which could be suitable for routine screening of microdeletions on the Y chromosome in azoospermic and oligozoospermic male infertility patients.

METHODSFive multiplex sets were established. Eighty-seven azoospermic and oligozoospermic patients undergoing intracytoplasmic sperm injection (ICSI) in the in vitro fertilization (IVF) center and 30 azoospermic men undergoing testicular biopsy in the clinic of Urology Surgery were screened for microdeletions of Y chromosome.

RESULTSA total of 19 (16.2%) cases of microdeletions were found in 117 azoospermic and oligozoospermic patients by screening of Y chromosome microdeletions. Of these, 11 cases (18.0%) were found in 61 oligozoospermic patients, and 8 cases (14.3%) were found in 56 azoospermic patients.

CONCLUSIONThe multiplex PCR protocol presented in this study is an easy-to-do and reliable method for detecting microdeletions on the Y chromosome. Routine screening of microdeletions on the Y chromosome for azoospermic and oligozoospermic patients is essential.

Chromosome Deletion ; Chromosomes, Human, Y ; genetics ; Female ; Genetic Testing ; methods ; Humans ; Infertility, Male ; diagnosis ; genetics ; Male ; Polymerase Chain Reaction

OBJECTIVETo study the outcome of hepatitis C virus (HCV) vertical transmitted infants.

METHODSThirteen HCV vertical infected infants were followed up for 10 years. HCV antibody and HCV RNA in the blood samples from them were tested using second generation HCV antibody EIA kits and RT-PCR, respectively.

RESULTSAmong the 13 infants, one developed clinical hepatitis C, and serum HCV antibody and HCV RNA could be detected for 7 and 8 years, respectively. Three were subclinical hepatitis C, serum HCV antibody continued to be positive for 12 months (2 infants) and 24 months (1 infant), respectively, and serum HCV RNA turned to be negative at the 24th month (2 infants) and the 60th month (1 infant), respectively. Nine were HCV insidious infection, whose serum HCV antibody and HCV RNA turned to be negative in 12 months. During the eight to ten years, there was no infants with anti-HCV or HCV RNA positive again.

CONCLUSIONSIt is rarely happened that vertical transmitted HCV induce chronic HCV carrying state and chronic viral hepatitis, and most of the infected infants have good outcome.

Child ; Child, Preschool ; Female ; Hepatitis C ; transmission ; Hepatitis C Antibodies ; blood ; Humans ; Infant ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; Pregnancy ; Time Factors

BACKGROUNDThe initial classic classification of duplex kidney into complete (two ureters) and incomplete ("Y" shaped ureter) types are based on the ureter status. At the meantime, the features of the upper and lower moieties of duplex kidney were very crucial for appropriate procedure of hemi-nephrectomy, which was most commonly used for addressing the issues caused by a duplex kidney; and recently more applications of laparoscopy were used. In this study, we aimed prudently to propose a new classification based on the features of the upper and lower moieties of duplex kidney.

METHODSSixty-five children with 83 duplex kidneys were reviewed retrospectively. Based on kidney morphology found in CT urography and surgical findings, duplex kidney was classified into five types.

RESULTSThe first was the appendant type (36/83) and its feature was that the mini upper moiety was located on top of the lower one, with a visualized shallow groove between them. The ureter was dilated with an ectopic orifice or ureterocele. The second was the embedded type (13/83), the feature of which was that mini upper moiety located in the interior top of the lower one within the same capsule. The upper ureter was dilated with an ectopic orifice or ureterocele. The third was the hydronephrosis type (12/83). The severe hydronephrotic upper moiety was almost as big as the lower moiety. The upper ureter was severely dilated and circuitous with an ectopic orifice. The forth was the dual-poor type (2/83). The two moieties were all very small with "Y" shaped ureters and ectopic orifices. The last was the dual-well type (20/83). The upper moiety was almost the same size as the lower one, without apparent dilation of "Y" shaped or double ureters.

CONCLUSIONBased on kidney morphology, duplex kidney can be mainly classified into five types which can be depicted by CT urography prior to management and can provide an aid in selecting a successful course of surgical correction.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Kidney ; abnormalities ; diagnostic imaging ; Kidney Diseases ; diagnosis ; diagnostic imaging ; Male ; Radiography ; Retrospective Studies

OBJECTIVETo explore the possibility of creating a rat new scar model by inserting gelatin sponge into rat excisional wounds.

METHODSTwo full-thickness wounds were created in each of total 49 SD rats. In the Experimental group (n = 19), a regular incisional wound (1 cm) was created on the left side, and an excisional wound of 1.0 cm x 0.2 cm was created on the right side with a gelatin sponge inserted. In control 1 group (n = 15), an excisional wound with sponge insertion was created on both sides of rats. In control 2 group (n = 15), two excisional wounds were created on both sides, and only one side wounds were inserted with a sponge. Animals were sacrificed at various time points for different examinations.

RESULTSThe wound/scar width increased 4 - 11 times in inserted wounds than in regular incisional wounds (P < 0.01), with an obvious delay of epithelialization. No difference in wound/scar width was found in both sides of wounds of control 1 group at various locations. In contrast to the linear scar of sponge-inserted wounds, contracted and irregular scar was found in non-inserted wounds of control 2 group.

CONCLUSIONSGelatin sponge insertion can create a thick linear scar in rat wounds, and thus provides a new model for scar research.

Animals ; Cicatrix ; pathology ; Dermatologic Surgical Procedures ; Disease Models, Animal ; Gelatin Sponge, Absorbable ; Male ; Rats ; Rats, Sprague-Dawley ; Skin ; pathology ; Suture Techniques ; Wound Healing

OBJECTIVETo study the effect of Itk down regulation on Jurkat cell proliferation and inflammatory cytokines production, and provide useful data for Itk as an attractive target for potential drugs.

METHODSThree shRNAs against different region of Itk were constructed and cotransfected with pEGFP-C1-hItk. The shRNA, which can knock down Itk, was selected and packed into lentivirus. After Jurkat cells were transfected with shRNA lentivirus, the change of Itk protein expression, cell proliferation and cytokines production was observed.

RESULTSItk mRNA was reduced about 55% in Jurkat cells transfected with Itk-shRNA1, compared with that in control cells shRNAnon (P < 0.05). Knocking down Itk expression had a profound inhibitory effect on Jurkat cell proliferation. In addition, there was a substantial decrease in level of cytokines, such as IL-2, IL-5, IL-10 and IFN-gamma, produced by cell transfected with Itk-shRNA1.

CONCLUSIONKnocking down Itk expression can inhibit Jurkat cell proliferation and inflammatory cytokines production.

Animals ; Cell Proliferation ; Cytokines ; genetics ; immunology ; Down-Regulation ; Gene Knockdown Techniques ; Humans ; Interleukin-10 ; genetics ; immunology ; Interleukin-2 ; genetics ; immunology ; Interleukin-5 ; genetics ; immunology ; Jurkat Cells ; cytology ; enzymology ; immunology ; Mice ; Protein-Tyrosine Kinases ; genetics ; immunology