Th17/Treg cell expression in children with primary nephritic syndrome and the effects of ox-LDL on Th17/Treg cells.

To investigate the role of T-helper cells/Treg (Th17/Treg) and morbidity factors related to primary nephritic syndrome (PNS) in children, as well as the influence of ox-low density lipoprotein (ox-LDL) on Th17/Treg expression in children with PNS. To clarify the pathogenesis of PNS in children, 50 children with PNS treated in our hospital were enrolled in the study group. Additionally, 20 healthy children who came to our hospital for physical examination during the same period were enrolled in the control group. Th17 and Treg cells in children belonging to the two groups were detected by flow cytometry; the numbers of Th17/Treg cells in peripheral blood mononuclear cells at different concentrations of ox-LDL were detected simultaneously. Ox-LDL can affect the number of Th17/Treg cells in peripheral blood mononuclear cells, and both cell types decreased with increasing concentration of ox-LDL, with the numbers being significantly lower in the control group. However, the decrease in the number of Th17 cells was statistically insignificant (P > 0.05), whereas the decrease in Treg cells was more obvious and statistically significant (P < 0.05). The effect of ox-LDL the number of Treg cells was stronger than that on Th17 cells. We concluded that the imbalance of Th17/Treg cells influenced by high and low ox-LDL concentrations in children with PNS might be the immunological basis of the disease.

Association of the VRK2 gene rs3732136 polymorphism with schizophrenia in a Northwest Chinese Han population.

Previous studies have found that the vaccinia related kinase 2 gene (VRK2) polymorphism was associated with schizophrenia (SCZ) in the worldwide population. This association was further supported by VRK2 mRNA expression patterns and brain structure variations. Here, we analyzed four single nucleotide polymorphisms (SNPs) of the VRK2 gene in a total population of 893 samples, consisting of 360 patients with SCZ and 533 healthy controls of Han Chinese descent using the SNPscan method. Single SNP, haplotype, and gender-specific association analyses were performed. We found that rs3732136 was significantly associated with SCZ (P = 0.042; odds ratio = 1.25; 95% confidence interval = 1.01-1.55). Further genotype and haplotype association analyses suggested a similar pattern. Our data provide preliminary evidence that the VRK2 gene might play a major role in the development of SCZ in the Northwest Chinese Han population.

Dopamine receptor D1 but not D3 essential for morphine-induced conditioned responses.

Recent studies indicate the involvement of dopamine receptors D1 and D3 in the regulation of locomotor stimulant and conditioned responses to morphine in mice. Moreover, expression of brain-derived neurotrophic factor (BDNF) may be modulated by D1 and D3 receptor activities in the nucleus accumbens (NAc) and prefrontal cortex (PFC). However, the underlying interactions between D1 and D3 receptors and BDNF in the expression of behavioral responses controlled by drug-associated cues have not yet been fully elucidated. In this study, we used dopamine receptor mutant mice to explore the roles of the D1 and D3 receptors in locomotion and morphine-induced place preference; furthermore, we investigated the effects of morphine on BDNF expression in the NAc and PFC of the mouse brain. Our results show that D1 receptor but not D3 receptor mutant mice had decreased sensitivity to acute morphine-induced (10 mg/kg) locomotion (D1: 3814.82 ± 319.9 cm vs D3: 8089.64 ± 967.4 cm). Furthermore, D1 receptor mutant mice did not acquire morphine-conditioned place preference (D1: -18.3 ± 59.9, D3: 217.7 ± 64.1) and showed decreased BDNF expression in the NAc (D1: 0.33 ± 0.07 fold, D3: 2.21 ± 0.18 fold) and PFC (D1: 0.74 ± 0.15 fold, D3: 1.68 ± 0.22 fold) compared with wild-type and D3 receptor mutant mice. These findings suggest that the D1 receptor is necessary for the induction of cue-associated morphine seeking and modulates locomotor habituation processes in response to acute morphine. The dopamine receptor D1 but not the D3 is also critical for morphine-induced BDNF expression in the NAc and PFC.

Polymorphisms in the endothelial nitric oxide synthase gene associated with recurrent miscarriage.

Endothelial nitric oxide synthase (eNOS) is an enzyme that influences placental human chorionic gonadotropin production during gestation. Previous studies have indicated an association between eNOS activity, implantation, and maintenance of pregnancy, but proposed associations between polymorphisms of the eNOS gene and recurrent miscarriage (RM) are controversial. To identify markers contributing to the genetic susceptibility to RM, we examined the potential association between RM and 8 single nucleotide polymorphisms (SNPs; rs1799983, rs2070744, rs11771443, rs3918188, rs2853796, rs7830, rs1541861, and rs2853792) of the eNOS gene using the MassARRAY system (Sequenom, USA). The enrolled participants included 192 RM patients and 201 women with normal fertility as controls. The results showed that rs1799983 at exon 7 of the eNOS gene was significantly associated with RM (genotype: chi-square = 15.071, P = 0.001; allele: chi-square = 6.250, P = 0.016). Another significant association was observed for rs11771443 in the promoter (genotype: chi-square = 6.259, P = 0.044; allele: chi-square = 7.076, P = 0.008). Furthermore, strong linkage disequilibrium was observed in 3 blocks (D' > 0.9), and significantly fewer T-T-G haplotypes (chi-square = 5.981, P = 0.015) residing in block 1 were found in RM patients. These findings point to a role for eNOS gene polymorphisms in RM in the Chinese Han population and may be informative for future genetic or neurobiological studies of RM.

Nine polymorphic STR loci in the HLA region in the Shaanxi Han population of China.

A large number of microsatellite genetic markers have been identified in the human leukocyte antigen (HLA) region. We investigated genetic polymorphism of the nine short tandem repeat (STR) loci (D6S276, MOGCA, D6S265, MIB, D6S273, G51152, TAP1CA, RING3CA, and D6S291) in the HLA region in the Shaanxi Han population. Using a fluorescence-labeled multiplex-PCR STR typing method, 6-13 alleles were detected in these nine STR loci in 150 unrelated Han Chinese from the region of Shaanxi, China. The distributions of the genotypes at these nine loci were in Hardy-Weinberg equilibrium. We conclude that these nine STR loci have a high level of genetic polymorphism; they would be useful for population genetic studies, pre-transplantation HLA typing, forensic and paternity testing, etc.

A study on the relationship between TCTA tetranucleotide polymorphism of the HPRT gene and primary hyperuricemia.

We examined polymorphism of the TCTA tetranucleotide sequence in the 3rd intron of the hypoxanthine-guanine phosphoribosyltransferase (HPRT) gene in the Han population of Ningxia Province in China. We also looked for a possible relationship between STR polymorphism in the 3rd intron of the HPRT gene and primary hyperuricemia. We used Chelex-100 to extract DNA, then PCR, PAGE and silver staining for allele genotyping and DNA sequencing to obtain the distribution of the alleles. We found, for the first time, that there is high STR polymorphism in the 3rd intron of the HPRT gene. We detected 5 STR alleles in this intron in the Han population of Ningxia Province, with 15 genotypes in females; significant differences were observed in the distribution of alleles and genotypes between control and patient groups for both males and females. Alleles of the TCTA repeat in the 3rd intron of the HPRT gene were found to be associated with primary hyperuricemia; consequently, these alleles may be considered risk factors for primary hyperuricemia.

Polymorphism G861C of 5-HT receptor subtype 1B is associated with heroin dependence in Han Chinese.

The serotoninergic (5-HT) system regulates neuronal activity in broad brain regions, and appears to be particularly important for modulating behavioral and physiological functions such as mood, emotion, sleep and appetite. Central 5-HT deregulation may be involved in many neuropsychological disorders, which include substance abuse and addiction. Previous studies suggest that genetic polymorphisms in some 5-HT receptor genes may relate to heroin dependency. Here we examined potential association between heroin dependence and four single nucleotide polymorphisms (SNPs) of 5-HT receptors (A-1438G and T102C of HTR(2A), and G861C and A1180G of HTR(1B)) in a cohort of Han Chinese. Participants included 303 heroin-dependent subjects who were recruited into the Methadone Maintenance Treatment (MMT) Program in the Xi'an Mental Health Center, and 300 healthy controls. The resulting data yielded a significantly higher frequency of the HTR(1B) G allele with G861C among the heroin-dependent subjects relative to controls (p=0.001 after Bonferroni correction). Further genotype and clinical phenotype correlation study of the G861C carriers showed that the amount of heroin self-injection was higher in patients with the GG genotype relative to CC and CG genotypes (p<0.01). These findings point to a role for HTR(1B) polymorphism in heroin dependence among Han Chinese, and may be informative for future genetic or neurobiological studies on heroin dependence.

Association between heroin dependence and prodynorphin gene polymorphisms.

Dynorphin peptides and k-opioid receptor are important in the rewarding effects of drugs of abuse such as heroin. This study examined potential association between heroin dependence and four single nucleotide polymorphisms (SNPs) of prodynorphin (PDYN) gene (rs35286281 in promoter region and rs1022563, rs2235749, rs910080 in 3'UTR). Participants included 304 heroin-dependent subjects and 300 healthy controls. Genotype, allele frequencies and difference between groups were analyzed by HaploView 4.0 and SPSS 11.5 software. The analysis indicated a significant higher frequency of the PDYN 68bp VNTR (rs35286281) H allele in heroin-dependent subjects than in controls (p=0.002 after Bonferroni correction). Strong linkage disequilibrium was observed between rs1022563, rs2235749 and rs910080 polymorphism (D'>0.9). Significantly more TCT haplotypes were found in heroin-dependent patients than in the controls (p=0.006 after Bonferroni correction). We found significant pointwise correlation of these three variants (rs1022563, rs2235749 and rs910080) with heroin dependence. These findings support the important role of PDYN polymorphism in heroin dependence, and may guide future studies to identify genetic risk factors for heroin dependence.

Dopamine D1 but not D3 receptor is critical for spatial learning and related signaling in the hippocampus.

Experimental evidence suggests the involvement of the brain dopaminergic system in learning and memory processes, although the associated molecular mechanism has yet to be fully characterized. Memory formation occurs via a number of signaling pathways associated with activation of many synaptic plasticity-related proteins, including the N-Methyl-D-aspartic acid (NMDA) receptor, Ca(2+)/calmodulin-dependent protein kinase II (CaMKII), mitogen-activated protein kinases (MAPKs) and the cAMP-response element binding protein (CREB). To evaluate the roles of dopamine D(1) and D(3) receptors in spatial learning and memory and underlying molecular events, we have used genetically modified mice carrying either the D(1) or D(3) receptor gene mutations to explore the intracellular signaling pathways using Morris water maze (MWM) tasks. We show that D(1) receptor mutant mice do not acquire spatial memory and do not show hippocampal activation of extracellular signal-regulated kinase (ERK) compared to wild-type mice. D(3) receptor mutant mice exhibit apparent normal learning abilities in the MWM test and normal activation of MAPK signaling. Furthermore, activation of the NMDA receptor R1 subunit (NR1), CaMKII and CREB in the hippocampus is also significantly lower in D(1) receptor mutant mice compared to wild-type and D(3) receptor mutant mice. These results suggest that dopamine D(1) but not D(3) receptor is critical for spatial learning. D(1) receptor-mediated signaling, associated with activation of NR1, CaMKII, ERK and CREB, is highly involved in the encoding of spatial learning and memory.

Distribution of human leukocyte antigen alleles and haplotypes in Oroqen and Ewenki nationality minority in Inner Mongolia Autonomous Region of China.

The frequencies of the human leukocyte antigen alleles HLA-A,-B, DRB1 and the A-B, A-DRB1, B-DRB1, A-B-DRB1 haplotypes were investigated through means of PCR-based reverse line-strip sequence specific oligonucleotide hybridization on 108 Oroqen and 104 Ewenki nationality unrelated healthy individuals from the Inner Mongolia Autonomous Region of China. A total of thirteen different HLA-A alleles, 21 different HLA-B alleles and 13 different HLA-DRB1 alleles were detected in the Oroqen ethnic group and the most frequent HLA alleles found were A*24(35.65%), B*15(17.92%), and DRB1*09(17.59%), respectively. The common HLA-A-B-DRB1 haplotypes were A*24-B*40-DRB1*09(5.09%), A*24-B*48-DRB1*12(2.78%) and A*24-B*51-DRB1*04(2.78%); and the HLA-A*33-B*58, A*30-B*13, A*01-B*37, A*33-DRB1*03, A*01-DRB1*10, A*30-DRB1*07, B*37-DRB1*10, B*58-DRB1*03, B*38-DRB1*08, B*13-DRB1*07 were significant positive linkage disequilibrium in the Oroqen nationality group. In total, 14 different HLA-A alleles, 27 B alleles and 12 DRB1 alleles were found in Ewenki nationality group, and the most frequent HLA alleles found were A*24(24.49%), B*40(17.35%), and DRB1*04(14.80%), respectively. The common HLA-A-B-DRB1 haplotypes were A*33-B*58-DRB1*03(6.25%), A*01-B*51-DRB1*11(2.88%) and A*24-B*40-DRB1*09(2.88%); the HLA-A*33-B*58, A*29-B*44, A*03-B*52, A*33-DRB1*03, A*29-DRB1*07, A*24-DRB1*09, B*58-DRB1*03, B*08-DRB1*03, B*46-DRB1*09 were significant positive linkage disequilibrium in Ewenki nationality group. The distribution of HLA A,-B, DRB1, alleles haplotypes frequencies and phylogenetic tree indicated that the Oroqen and Ewenki population groups belongs to northern group of China, together as a group cluster.

Pharmacological treatment for heart failure: a view from the brain.

Systolic heart failure is a feed-forward phenomenon with devastating consequences. Impaired cardiac function is the initiating event, but central nervous system mechanisms activated by persistent altered neural and humoral signals from the periphery play an important sustaining role. Animals with experimentally induced heart failure have neurochemical abnormalities in the brain that, when manipulated, profoundly affect sympathetic drive, volume regulation, and cardiac remodeling--critical determinants of outcome. This brief review explores recent studies that provide a strong rationale for the development of pharmaceutical agents that target central nervous system abnormalities in heart failure.

Central mineralocorticoid receptor blockade improves volume regulation and reduces sympathetic drive in heart failure.

The mineralocorticoid (MC) receptor antagonist spironolactone (SL) improves morbidity and mortality in patients with congestive heart failure (CHF). We tested the hypothesis that the central nervous system actions of SL contribute to its beneficial effects. SL (100 ng/h for 28 days) or ethanol vehicle (VEH) was administered intracerebroventricularly or intraperitoneally to rats with CHF induced by coronary artery ligation (CL) and to SHAM-operated controls. The intracerebroventricular SL treatment prevented the increase in sodium appetite and the decreases in sodium and water excretion observed within a week of CL in VEH-treated CHF rats. Intraperitoneal SL also improved volume regulation in the CHF rats, but only after 3 wk of treatment. Four weeks of SL treatment, either intracerebroventricularly or intraperitoneally, ameliorated both the increase in sympathetic drive and the impaired baroreflex function observed in VEH-treated CHF rats. These findings suggest that activation of MC receptors in the central nervous system plays a critical role in the altered volume regulation and augmented sympathetic drive that characterize clinical heart failure.

Progression of heart failure after myocardial infarction in the rat.

This study examined the early neurohumoral events in the progression of congestive heart failure (CHF) after myocardial infarction (MI) in rats. Immediately after MI was induced by coronary artery ligation, rats had severely depressed left ventricular systolic function and increased left ventricular end-diastolic volume (LVEDV). Both left ventricular function and the neurohumoral indicators of CHF underwent dynamic changes over the next 6 wk. LVEDV increased continuously over the study interval, whereas left ventricular stroke volume increased but reached a plateau at 4 wk. Plasma renin activity (PRA), arginine vasopressin, and atrial natriuretic factor all increased, but with differing time courses. PRA declined to a lower steady-state level by 4 wk. Six to 8 wk after MI, CHF rats had enhanced renal sympathetic nerve activity and blunted baroreflex regulation. These findings demonstrate that the early course of heart failure is characterized not by a simple "switching on" of neurohumoral drive, but rather by dynamic fluctuations in neurohumoral regulation that are linked to the process of left ventricular remodeling.

Neurohumoral regulation in ischemia-induced heart failure. Role of the forebrain.

Congestive heart failure (CHF) is characterized by neurohumoral excitation. Increased sympathetic drive and activation of the reninangiotensin-aldosterone system (RAAS), with vasoconstriction and volume retention, are hallmarks of the CHF syndrome. Treatment strategies have targeted the peripheral influences of these two systems, but have not addressed the central mechanisms that drive them. We monitored the development of CHF following coronary ligation in adult Sprague-Dawley rats. Left ventricular dysfunction characteristic of CHF was confirmed by echocardiography, and the CHF syndrome was validated by measurements of circulating hormones, sodium appetite, thirst, renal sodium and water retention, and renal sympathetic nerve activity (RSNA). In CHF rats, neuronal activity in the hypothalamic paraventricular nucleus (PVN), which mediates downstream effects of forebrain circumventricular organs, was increased and was inhibited by blocking components of the RAAS at the forebrain level. Forebrain (AV3V) lesions and intracarotid (forebrain directed) injections of agents (captopril, losartan, spironolactone) that block RAAS substantially attenuated the behavioral and physiological manifestations of CHF. Intravenous losartan and captopril, in doses that lower arterial pressure, increased RSNA. These findings demonstrate an important role for RAAS-activated forebrain mechanisms in CHF and suggest that the central neural mechanisms driving sympathetic nerve activity and volume retention may persist and promote the progression of CHF despite treatments directed toward the peripheral influences of RAAS.

[Effect of electrical stimulation of afferent renal nerve on arterial blood pressure, heart rate and vasopressin in rabbits].

The effect of electrical stimulation of afferent renal nerve (ARN) on cardiovascular response, the synthesis and release of vasopressin were studied in rabbits. During the course of the experiment, the pathway of ARN to central nerve system was also analyzed. The results showed that electrical stimulation of ARN elicited significantly decrease of mean arteral blood pressure and heart rate as well as inhibition of cervical sympathetic nerve activity. In the event of the above physiological changes, the AVP concentration in supraoptic nucleus (SON), paraventricular nucleus (PVN) and plasma was increased, but that in hypophysis was decreased. Injection of sodium nitropruside (SNP) or AVPa indicated that increase of AVP release was due directly to stimulation of ARN. Nodose ganglionectomy or transversal section of spinal cord at different levels suggested that the main afferent pathway of ARN to higher level of central nerve system entered into spinal cord at T5-L2.

Expression of the calpastatin gene segment during spermiogenesis in human testis: an in situ hybridization study.

Serum obtained from an infertile woman contained antibodies that agglutinate human sperm. The antibodies interacted with a sperm protein with an estimated M(r) of 17.5 kD. The cDNA coding the 17.5-kD protein was isolated from a human testis lambda gt11 expression library and identified as a segment of the calpastatin gene. Single-stranded 35S-labeled RNA probes were prepared from the calpastatin cDNA segment. Using the techniques of in situ hybridization, the calpastatin mRNA was located in spermatids of human testis. The results support a previous observation that the calpastatin segment is produced during spermiogenesis and suggest that transcription of the calpastatin gene occurred during the postmeiotic haploid stage of spermatogenesis.

[Reversal of malignant phenotype of human hepatoma cells by antisense c-ets-2, c-myc and N-ras].

A recombinant retroviral vector was constructed which expressed antisense RNA of c-ets-2, c-myc and N-ras. The pseudotype virus was packaged and rescued by transfection in PA317 cells and used to infect human hepatoma cell line SMMC-7721. After selection with G418, resistant colonies were obtained. Stable integration of retrovirus in infectants was shown by Southern hybridization of genomic DNA and the presence of antisense RNA was detected by RNA dot blot hybridization. It was demonstrated that the antisense RNAs did inhibit the growth of human SMMC-7721 hepatoma cells. The ability to form colony in soft agar and tumorigenicity in nude mice of SMMC-7721 were significantly suppressed by the antisense RNAs. The result implicates the potential value in future cancer gene therapy.

Calpastatin gene in human testis.

Antibodies present in a serum obtained from an infertile woman interacted with a 17.5 kD glycoprotein (BS-17 component) extracted from human sperm by Western blot. Polyclonal antibodies were raised against the BS-17 component and used to identify positive staining clones from a human testis lambda gt11 expression library. The cDNA encoding the BS-17 component was isolated and its nucleotide sequence determined. The BS-17 cDNA contained 758 nucleotides with an open reading frame of 558 nucleotides encoding a polypeptide consisting of 186 amino acids. The BS-17 cDNA showed 99.7% homology in 758 nucleotides overlap with the 3' terminus of the gene coding calpastatin and 99.5% identity in 186 amino acid overlap with the carboxyl terminus of calpastatin. The BS-17 component of human sperm corresponds to the carboxyl terminus of calpastatin. This conclusion is supported by the finding that the polyclonal antibodies also interacted with a 84 kD protein corresponding to the M(r) of calpastatin.

Fertility studies with antisperm antibodies.

Serum obtained from an infertile subject possessed antibodies that interacted with a human sperm glycoprotein with an estimated M(r) of 17,550 and pI of 5.65 containing 17.7% neutral hexoses and designated as the BS-17 component. Polyclonal antibodies raised against the BS-17 antigen blocked the capacity of human sperm to fertilize zona-free hamster ova in vitro; however, the antibodies did not influence the binding of human sperm to zone-free ova or alter the motility of human sperm. The antibodies inhibited the capacity of mouse sperm to fertilize ova upon in vivo insemination. The BS-17 antigen was detected in human, rat, mouse, rabbit, and hamster sperm by an immunocytochemical method, using polyclonal anti-BS-17 antibodies. Intense staining occurred over the surface of the acrosomal region of all mammalian sperm. The results suggest that the production of anti-BS-17 antibodies contribute to infertility by preventing the capacitation of sperm and/or by blocking the ability of capacitated sperm to fertilize the egg.

[Modulatory role of the cerebellar cortex during the classically conditioned nictitating membrane response in rabbit].

Classical conditioning of the rabbit nictitating membrane(NM) response was accomplished by standard delay procedure of paired tone (CS) and airpuff (US) stimuli. Upon reaching a criterion of 90% conditioned responses (CRs) in 3 consecutive blocks, an electrolytic lesion was made at the hemicerebellar VI cortical lobule on the training side. Results showed that the learned response of the dentate-interpositus (D-I) nuclei and the conditioned NM response were completely or almost completely eliminated without affecting the unconditioned response. However, cerebellar cortical lesion performed on animals being over 1 week overtrained produced no effect on CRs and the learned response of the D-I nuclei. Meanwhile, the spontaneous activity of the D-I nuclei during consolidation reduced in frequency as a result of the cortical lesion at the early stage of learning. Our findings indicate that the cerebellar cortex plays a modulatory role in the generation of the conditioned NM response and the development of the learned response of the D-I nuclei at an early stage of learning. With consolidation, the D-I nuclei become free of control from the cerebellar cortex and serve as the essential component of the memory trace for this model system.