Objective To explore wbether there is an association between the single nucleotide polymorphisms (SNPs) of phosphodiestierase-4A (PDE4A) and bipolar disorder in Han population.Methods A casecontrol association study was done in this study,432 bipolar disorder patients(patients group),and 569 age and gender-matched controls(control group)were recruited.Real-time fluorescent quantitative polymerase chain reaction (PCR) was used to detect genotypes and alleles distributive frequency of rs1051738 and rs7256672 of 432 bipolar disorder patients and 569 normal people.Results The PDE4A rs1051738 genotypes and alleles frequency distribution between patients group(AA,AC,CC genotypes:12％,8％,80％,A,C alleles:16％,84％) and control group (AA,AC,CC genotypes:2％,19％,79％,A,C alleles:12％,88％) showed statistically significance (P＜ 0.01).The frequency of PDE4A rs7256672 genotypes and alleles distribution between patients group(GG,GT,TT genotypes:18％,50％,32％,G,T alleles:43％,57％) and control group (GG,GT,TT genotypes:20％,48％,32％,G,T alleles:45％,55％)showed no significant difference(P=0.620,P=0.446).Conclusion The PDE4A rs1051738 polymorphism may be responsible for an increase in susceptibility to bipolar disorder,indicating that it maybe a functional site with marked significance to bipolar disorder.

Objectives To investigate the role of angiogenic T cells (Tang) and endothelial progenitor cells (EPC) in the pathogenesis of preeclampsia. To explore the relationship between Tang and EPC. Methods From Mar 2013 to Aug 2014, 40 patients diagnosed preeclampsia (PE) and delivered in Hunan Provincial People′s Hospital. A total of 20 of them were defined as the mild preeclampsia group and the other 20 cases were recruited as the severe preeclampsia group. And 24 healthy pregnant women wererecruited as the control group. The percentage of Tang and EPC in peripheral blood mononuclear cells (PBMC) were determinated by flow cytometry between 28 and 40 gestational weeks. Results (1) There was no significant difference in the age, pre-pregnancy body mass index(Pre-BMI) or gestational age among the three groups (P>0.05). The differences of blood pressure among the three groups were statistically significant (P<0.05). The gestational week at delivery, the birthweight of the neonates and the 1 minute Apgar score in the severe preeclampsia group were lower than those in the mild preeclampsia group and the control group, with statistically significant differences (P<0.05). The morbidity of neonatal asphyxia in the severe preeclampsia group was 35%(7/20);and in the mild preeclampsia group it was 5%(1/20), with statistically significant difference( P<0.05). (2) The percentage of Tang in maternal peripheral blood was(52.7 ± 8.0)%, (47.5 ± 8.8)% and (45.5 ± 8.7)% in the control group, the mild preeclampsia group and the severe preeclampsia group, respectively. The difference among the three groups was significant (F=4.248,P<0.05), and SNK q analysis showed there was significant difference between the control group and the severe preeclampsia group(P<0.05).While there was no statistically significant difference between the mild and the severe preeclampsia group, nor between the control group and the mild preeclampsia group(P>0.05). (3) The percentage of EPC in maternal peripheral blood was (0.16±0.07)%, (0.09±0.07)%and (0.08±0.05)%in the control group, the mild and the severe preeclampsia group, respectively. Analysis of variance showed that difference among the three groups was significant (F=9.351, P<0.05). The percentage of EPC in the mild or the severe preeclampsia group was significantly higher than that of the control group(P<0.05). (4) There was no statistically significant correlation between the Tang level and the EPC level in the control group ( r=-0.325, P>0.05). In the preeclampsia group (including mild and severe cases), there was positive correlation between the Tang level and EPC level (r=0.667, P<0.01). The positive correlation between Tang level and EPC level were proved respectively in the mild preeclampsia group (r=0.803, P<0.01) and the severe preeclampsia group (r= 0.520, P<0.05). Conclusions The number of Tang had some correlation with the pathogenesis of preeclampsia. The percentage of Tang had positive correlation with the level of EPC in women with preeclampsia. Tang might have some influence on the change of EPC′ level. Tang together with EPC were likely to contribute to the angiogenesis in preeclampsia.

AIM To study if the activated T lymphocytes induced by hepatocellular carcinoma (HCC) rejection antigenic peptide SLIVHLNEV (mutant peptide of C met) pulsed dendritic cells (DCs) can provide protection against tumor challenge in nude mice HCC model and in vitro . METHODS SLIVHLNEV was eluted from HCC cell line HHCC by mild acid buffer and was identified by the mass spectrometry technique. SLIVHLNEV pulsed DCs isolated from HLA A2+ ruptured spleen were co cutured with isogeneic T lymphocytes. Cytotoxicity was tested using lactate dehydrogenase (LDH) release assay. The induced cytotoxic T lymphocytes (CTLs) were implanted into nude mice in order to protect them against transplanted HHCC tumor cells. The inhibition effect of CTLs on the growth of implanted tumor in nude mice was also observed. RESULTS The CTLs induced by SLIVHLNEV pulsed DCs had unique ability to kill HHCC tumor cells in vitro. They also protected the nude mice against the further challenge of HHCC tumor cells and inhibited the growth of implanted tumor. CONCLUSION DC based HCC rejection antigenic peptide SLIVHLNEV vaccine can induce powerful immune reaction both in nude mice HCC model and in vitro.

Objective To observe the action of Rhodiola Crenulate(RC) oral liquid reproduction function for the male rats. Methods 100 SD rats were randomly divided into 5 groups with equal gender number respectively. The male rats were given RC oral liquid (0, 2.48 g/kg, 7.48 g/kg, 24.80 g/kg) and Nanbao (2.00 g/kg)by oral twice a day for 48 d, respectively. Then the male rat was put into the same box in which the female rat whit two-side ovarys were excised for 15 d, respectively. The intercoursing ability,testosterone levels, and the weight of immune organs of the male rats were examined. 40 four-week-old rats were divided into 5 groups. Except for blank control, all rats's testis were excised. The castrated rats were treated with RC oral liquid (2.48 g/kg, 24.8 g/kg)and Methyltestosterone (2.00 mg/kg). The blank control group and model control group were given the same dose NS by oral twice a day for 28 d, respectively. The weight of accessory sex organs in castrated rats was observed. Results The level of testosterone and the organ coefficients of thymus and spleen in RC oral liquid groups were higher than blank control (P<0.05 or P<0.01).The weight of accessory sex organs of castrated rats in RC oral liquid groups were heavier than model control (P<0.05). Conclusion RC oral liquid can improve the reproduction function in male rats.

Objective To explore whether there is association between the polymorphisms of serotonin 2A receptor(5-HT2A) rs6311,rs6305 and adenylate cyclase rs2230739 with episode of bipolar disorder.Methods A case-control association study was done in this study,152 bipolar disorder patients (patients group),and 187 ageand gender-matched controls (control group) were recruited.Ligase detection reaction(LDR) was used to detect the distributive frequency of rs6311,rs6305 and rs2230739 of 152 bipolar disorder patients and 187 normal people.Results The 5-HT2A receptor rs6311 genotypes and alleles frequency distribution between patients group(11,12,22 genotypes:21％,61％,18％ ; 1,2 alleles:51％,49％) and control group(11,12,22 genotypes:44％,45％,11％ ; 1,2 alleles:66％,34％) showed statistically significance (P＜0.01).There were no differences of 5-HT2A receptor rs6305 genotypes and alleles between patients group and control group.The frequency of rs2230739 genotypes and alleles distribution between patients group (11,12,22 genotypes: 20％,43％,37％ ; 1,2 alleles: 42％,58％) and control group(11,12,22 genotypes: 17％,48％,35％ ; 1,2 alleles: 41％,59％) showed no significant difference(P=0.661,P=0.846).Conclusion The 5-HT2A receptor rs6311 polymorphism may be responsible for an increase in susceptibility to bipolar disorder,indicating that it maybe an marker to bipolar disorder.

OBJECTIVE:To investigate the effect of Wuzhi soft capsule on the pharmacokinetics of sirolimus in rats. METH-ODS:Wistar rats were randomly divided into 5 groups,with 6 rats in each group. They were given very low-dose,low-dose,me-dium-dose and high-dose of Wuzhi soft capsule(67,134,268 and 536 mg/kg)ig or blank solvent,respectively;and then given si-rolimus 0.4 mg/kg. The blood 100 μl was sampled by capillary eyes before giving sirolimus(0 h)and 0.5,0.75,1,1.5,2,2.5, 3,4,6,8,12,24 and 36 h after giving sirolimus and put into edetic acid anticoagulation tube. Blood concentration of sirolimus was assayed by LC-MS/MS. The pharmacokinetic parameter was calculated by DAS 2.0 software using non-compartment model. RESULTS:The pharmacokinetic parameters of sirolimus in very low-dose,low-dose,medium-dose and high-dose groups and blank solvent group were cmax of(6.79±1.15),(17.40±3.13),(21.24±3.14),(22.06±4.82),(2.80±0.72)ng/ml;t1/2 of(11.01± 0.82),(12.20±2.02),(12.28±2.38),(12.36±0.73),(10.59±0.69)h;AUC0-∞ of(85.79±15.26),(162.18±41.75),(273.12± 73.69),(268.79±28.46),(36.72±3.01)ng·h/ml,respectively. Compared with blank solvent group,cmax and AUC0-∞ of sirolimus were all increased in very low-dose,low-dose,medium-dose and high-dose groups,but t1/2 changed slightly. CONCLUSIONS:Wu-zhi soft capsule can substantially enhance the absorption of sirolimus in rats.

Objective:To construct the prokaryotic expression vector of SEA mutant gene SEA(D227A).The gene was expressed in E.Coli, and the induced protein was purified and identified.Methods:The SEA gene was cloned by PCR from Staphylococcus aureus strain FRI 100.D227A was introduced by changing the Asp codon GAT into GCT(Ala)in the primer.The expression plasmid pRSET SEA(D227A)was constructed and transformed into E.Coli BL21(DE3)pLysS.The induced protein was identified by Western blot.Results:The nucleotide sequence of SEA(D227A)was found to be identical to the designed sequence. E.Coli BL21(DE3)pLysS contained pRSET SEA(D227A)can express a 32 kD protein which can specially bind with the anti SEA mAb.The induced protein was purified with Ni 2+ system.Conclusion:The SEA(D227A)gene was constructed and expressed successfully.The study gave a clue to the research of low toxic superantigen. [

Objective To evaluate MRI measurement of basal ganglia volumes in patients with Tourette's syndrome.Methods Ten patients with Tourette's syndrome(TS)and 10 healthy volunteers were studied.Volumes of bilateral candate,putamen and pallidum were measured,and the results were analyzed using paired t test.The basal ganglia volume was normalized according to individual brain volume.The basal ganglia volumes of TS patients were compared with normal control group using independent-sample t tesL Results In 10 healthy volunteers,volumes of the left caudate,putamen,pallidum were significantly larger compared with those of the right side(P＜0.05).However,there were no significant differenees between bilateral basal ganglia volumes(P＞0.05)in TS patients.After normalized processing,the volumes oftlle left candate(7.06±0.48)cm3,putamen(8.81 ± 1.01)cm3,pallidum(2.64 ± 0.38)cm3were smaller than those of control group[caudate(11.05±1.86)cm3,putamen(9.97 ±1.11 ) cm3, paUidum (3.04 ± 0.37 ) cm3 ] (t= - 6.577, - 2.457, - 2.376, P＜0.05 ).The volume of the right caudate (7.32 ± 0.26) em3 in "IS patients was significantly smaller compared with the control group (9.81±1.83) cm3 (t = -4.258, P ＜0.01 ).However, the volumes of the right putamen and pallidum had no significant difference between two groups (P＞0.05).Conclusion The basal ganglia volumes were significantly decreased in patients with TS.MRI volumetric measurement was an important tool for evaluating pathologic changes of TS.

OBJECTIVETo explore the protective mechanism of electroacupuncture (EA) at "Neiguan" (PC 6) on ischemic myocardial injury, and to explain the response patterns and characteristics of the specific effect of acupoints along meridians in sodium channel in the level of cardiac organ.

METHODSA total of 60 SPF male rats were randomly divided into a blank group, a model group, a non-acupoint group, a Neiguan group and a Lieque group, 12 cases in each one. Except the blank group, rats in the remaining group were treated with subcutaneous injection of isoprenaline to establish the model of myocardial ischemia. Rats in the Neiguan group, Lieque group and non- acupoint group were treated with EA, dilatational wave, with a frequency of 2 Hz/20 Hz. The intensity was 2-3 mA. The needles were retained for 20 min per time, once a day for consecutive 7 days. In the blank group and control group, the rats were grasped and fixed at the treating time each day. The western-blot method was used to test the expression of voltage-gated sodium channel alpha subunit (Nav 1.5), protein tyrosine kinase (PTKs) and protein tyrosine phosphatase (PTPs).

RESULTSThe expression of Nav 1.5 and PTKs in the model group was lower than that in the blank group (both P<0. 01); the expression in the Neiguan group and Lieque group was higher than that in the model group (all P < 0.01); the expression of Nav 1.5 and PTKs in the Neiguan group was higher than that in the Lieque group (both P < 0.01). The expression of PTPs in the model group and non-acupoint group was higher than that in the blank group (both P < 0.01); the expression of PTPs in the Neiguan group and Lieque group was significantly down-regulated, which was lower than the model group (both P < 0.01); the down-regulation in the Neiguan group was significantly different from that in the Lieque group (P < 0.05).

CONCLUSIONEA at "Neiguan" (PC 6), by down-regulating the expression of PTPs, up-regulating the expression of Nav 1.5 and PTKs, is likely to achieve the aim of regulation on sodium channel activity and calcium overload, further to improve myocardial ischemia, which provides experimental basis for the theory of the specific effect of acupoints along meridians.

Acupuncture Points ; Animals ; Disease Models, Animal ; Electroacupuncture ; Humans ; Male ; Myocardial Ischemia ; genetics ; metabolism ; therapy ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sodium Channels ; genetics ; metabolism

OBJECTIVETo observe the effects and duration of electroacupuncture on the mechanical pain threshold induced by paclitaxel and explore its analgesic mechanism.

METHODSSixty-four C57BL/6J male mice were randomly divided into 4 groups, a normal+sham EA group, a normal+EA group, a medicine+sham EA(Med+ sham EA) group, a medicine + EA (Med + EA) group, 16 cases in each group. The model of chemotherapy-induced peripheral neuropathy was established with paclitaxel intraperitoneal injection on the 1st, 3rd, 5th, 7th day in the Med + sham EA group and the Med + EA group. EA of 30 min was used on bilateral "Zusanli (ST 36)" on the 9th, 11th, 13th, 16th, 18th, 20th, 23rd, 25th, 27th, 30th day in the EA groups, 2 Hz/100 Hz and 1~ 1.5 mA. Acupuncture was applied on the same acupoint at the same times in the sham EA groups. Mechanical pain thresholds were tested by VonFrey before and after model establishment, namely on the 8th, 14th; 21st and, 28th day. The heart blood of 8 mice was drawn quickly to collect serum in every group on the 31st day, and the contents of tumor necrosis factor α (TNF-α), interleukin-1α (IL-1α), interleukin-1β (IL-1β) in proinflammatory cytokine were examined by ELISA. Mechanical pain thresholds were tested by VonFrey for the rest 8 mice of each group until there was no apparent difference in the two paclitaxel groups once a week,namely on the 35th, 42nd, 49th day.

RESULTSThe pain thresholds of each group were not statistically different before model establishment (P > 0.05). After model establishment (on the 8th day), thresholds of the paclitaxel groups were lower than those of the normal groups (all P < 0.05). After EA, the mechanical pain thresholds of the Med + EA group were higher than those of the Med + sham EA group at all the time points, and there was statistical difference on the 14th, 21st and 28th day (all P < 0.05). The analgesic effect was lasting to the 49th day. The contents of TNF-α, IL-1α, IL-1β of the Med + EA group were decreased than those of the Med+sham EA group in different degree, with statistical significance of IL-1α (P < 0.05).

CONCLUSIONEA can effectively treat paclitaxel-induced peripheral neuropathy,and the analgesic mechanism is probably related to decreasing the proinflammatory cytokine.

Acupuncture Points ; Animals ; Antineoplastic Agents ; administration & dosage ; adverse effects ; Electroacupuncture ; Humans ; Interleukin-1beta ; genetics ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Neoplasms ; drug therapy ; Peripheral Nervous System Diseases ; etiology ; genetics ; metabolism ; therapy ; Tumor Necrosis Factor-alpha ; genetics ; metabolism