Objective To study expression of hypoxia inducible factor 1? (HIF 1?) in the lungs of hypoxic rats and to explore the role of HIF 1? in the pathogenesis of hypoxic pulmonary hypertension Methods Twenty Wistar rats were randomly divided into control group and hypoxia group The models of hypoxic pulmonary hypertension were established by exposure to hypoxia for 4 weeks according to our laboratory protocol Digoxin labelled cRNA probe for HIF 1? was prepared by in vitro transcription Northern blot was performed by using the HIF 1? cRNA probe and In situ hybridization was also conducted with rat lung tissue sections Results Northern blot hybridization showed minimally positive in the lung tissue of control group ,but strongly positive in hypoxia group In situ hybridization analysis with hypoxic rat lung tissue revealed that HIF 1? mRNA was expressed in bronchial epithelial cells (strongly positive reaction) and peribronchial proliferative lymphomatic tissue (weakly positive reaction),casually in alveolar wall cells (weakly positive reaction),but not in pulmonary arterial endothelium and smooth muscle cells Conclusions Our data suggested that chronic hypoxia could induce pulmonary hypertension characterized by pulmonary vascular remodeling HIF 1? mRNA expression was elevated in the lungs of rat under hypoxic condition HIF 1 may be involved in the pathophysiological changes in response to hypoxia and play an important role in the development of hypoxic pulmonary hypertension

Objective To explore the association of the calcitonin receptor (CTR) allelic polymor- phism in the 1377 bp region with the risk of idiopathic hypercalciuria (IH) in the Han nationality in Hubei area,and to study the pathogenesis of IH.Methods The CTR genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism in 76 patients with IH and 126 healthy controls from the Han nationality in Hubei area,using restriction endonuclease AluI.Results The distribution frequen- cies of AluI alleles in the 2 groups followed the Hardy-Weinberg equilibrium.The distribution frequencies of the CC,TC and TT genotypes were 73.7% ,17.1% and 9.2% in IH patient group,and 89.7% ,9.5% and 0.8% in control group;the distribution frequencies of C and T alleles in the 2 groups were 84.2% ,15.8% and 94.4% ,5.6% ,respectively.The distribution frequencies of T and TT alleles were higher,while those of C and CC alleles were lower,compared with control group;the differences between the 2 groups were signifi- cant (P＜0.05).Conclusions The results indicate that the C/T single nucleotide polymorphism in the CTR gene play a significant role in the mechanism of IH in the Han nationality in Hubei area in China.

The paper is aimed to identify SNP in Sarcandra glabra and Chloranthus spicatus, and authenticate S. glabra from Ch. spicatus and the mixture by using PCR amplification of specific alleles. SNPs in the ITS sequences of S. glabra and Ch. spicatus were found by ClustulX 2. 1 program and Bioedit software. Primers for authentic S. glabra and Ch. spicatus was designed according to the SNP site, and ITS sequence universal primers plus to the authentic primer to construct a multi-PCR reaction system, and then optimized the PCR reaction system. Five hundred and eighty band special for S. glabra and 470 bp band special for Ch. spicatus were found by using multi-PCR reaction. The multi-PCR reaction system could be applied to identify S. glabra and Ch. spicatus's leaves.
DNA, Plant ; analysis ; genetics ; DNA, Ribosomal ; genetics ; DNA, Ribosomal Spacer ; analysis ; genetics ; Magnoliopsida ; classification ; genetics ; Plant Leaves ; genetics ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; RNA, Ribosomal ; genetics ; RNA, Ribosomal, 18S ; genetics ; RNA, Ribosomal, 5.8S ; genetics ; Species Specificity

The study is aimed to assess the genetic diversity and genetic relationship of 18 Sarcandra glabra resources from different populations,and guide parent selection of cross breeding between these resources. The molecular marker technique ISSR was used to investigate the genetic diversity of the 18 resources. Data was analyzed by POPGEN 32, and a cluster diagram was presented by UPGMA. One hundred and ninety-eight amplified fragments were obtained using 23 ISSR primers. One hundred and eighty-four polymorphic loci were identified. Nei's genetic diversity index (h) was 0.32, Shannon diversity index (I) was 0.485 4. The genetic similarity coefficient among the resources ranged from 0.383 8 to 0.878 8 in an average of 0.661 2. The genetic distance between sample S2 and sample S18 was the farthest, so as between sample S3 and sample S18 both Nei's genetic distance was 0.957 5, The genetic distance between sample S4 and sample S5 was the closest, the Nei's genetic distance was 0.129 2,and the sample S1, S2, S3, S7, S10 were significantly different from the others based on the clustering analysis, the three groups S2 vs S3, S2 vs S6, S2 vs S18 were the best parent group selection. There was a middle level of genetic differentiation in the resources. The genetic distance between resources gives useful information to guide parent selection of cross breeding.
Conservation of Natural Resources ; DNA Primers ; genetics ; Genetic Variation ; Magnoliopsida ; classification ; genetics ; Microsatellite Repeats ; Phylogeny

OBJECTIVETo investigate the precise locations of the blood vessels and nerves surrounding the seminal vesicles (SV) in men and provide some anatomical evidence for SV-related minimally invasive surgery.

METHODSWe observed the courses and distribution of the blood vessels and nerves surrounding SVs and obtained the data for positioning the SV neuroplexes in 20 male pelvises.

RESULTSOne branch of the neuroplexes was distributed to the SVs bilaterally with the neurovascular bundles, (2.85 ± 0.18) cm from the median sulcus of the prostate (MSP), while another branch ran through the Denonvillier fascia behind the SV, (0.81 ± 0.06) cm from the MSP. The arterial SVs (ASV) originated from the inferior vesical artery and fell into 4 types, 55% going directly to the SVs as one branch, 15% running between the SV and the ampulla of the deferent duct as another branch, 25% downward as 2 branches to the SV and between the SV and the ampulla of the deferent duct respectively, and 5% as the other ASVs. The shortest distance from the ASV through the prostatic neuroplexus to the posterior SV was (1.08 ± 0.09) cm.

CONCLUSIONIn SV resection, neuroplexus injury can be reduced with a bilateral distance of < 2.85 cm and a posterior distance of < 0.81 cm from the MSP, and so can bleeding by vascular ligation between the SV and the ampulla of the deferent duct.

Biopsy ; Humans ; Male ; Prostate ; blood supply ; innervation ; Seminal Vesicles ; blood supply ; innervation ; Vas Deferens ; blood supply ; innervation

Objective To estimate the effect of glycyrrhetinic acid on epidermal growth factor (EGF)-induced proliferation of HaCaT cells,and to investigate its possible mechanism.Methods Methyl thiazolyl tetrazolium (MTT) assay was used to evaluate the proliferation of HaCaT cells treated with different concentrations of EGF (0,1,5,10,25,50,100 μg/L) and glycyrrhetinic acid (0,0.1,1.0,10,25,50,100μmol/L) alone,or the combination of 25 μg/L EGF with 25 μ mol/L glycyrrhetinic acid or 10 μ mol/L U0126 (an inhibitor of MEK1/2).Western blot was carried out to measure the protein expression of proliferating cell nuclear antigen (PCNA),Notch-1,ERK 1/2 and phosphorylated ERK 1/2 in HaCaT cells treated with 25 μg/L EGF,10 μmol/L U0126,25μmol/L glycyrrhetinic acid alone or in combination.Data were statistically analyzed by using t test,analysis of variance and correlation analysis with SPSS 17.0 software.Results EGF of 0-100 μg/L promoted the proliferation of HaCaT cells in a dose-dependent manner (r =0.798,P ＜ 0.05),and there was a linear correlation between the effect and concentration within the concentration range 0-50 μg/L (r =0.859,P ＜ 0.05).However,glycyrrhetinic acid of 10-100 μmol/L inhibited the proliferation of HaCaT cells in a dose-dependent manner (r =-0.945,P ＜0.01),and 10 μmol/L glycyrrhetinic acid could suppress the EGF (25 μg/L)-induced proliferation and phosphorylation of ERK1/2 in HaCaT cells.Also,both 25 μmol/L glycyrrhetinic acid and 10 μmol/L U0126 could attenuate the increase in PCNA and Notch-1 expression in HaCaT cells induced by 25 μg/L EGF.Conclusion Glycyrrhetinic acid can inhibit the EGF-induced proliferation of HaCaT cells,likely by suppressing the activation of ERK1/2 signaling pathway.

Objective To evaluate the effect of curcumin on the proliferation of and apoptosis in HaCaT cells induced by tumor necrosis factor α (TNF-α).Methods HaCaT cells were cultured with the presence of different concentrations (0,1,5,10,25,50,100 ng/ml) of recombinant TNF-α,curcumin of 20 μmol/L,or the combination of recombinant TNF-α (25 ng/ml) and curcumin (20 μmol/L),for 24 hours followed by the determination of cell proliferation with methyl thiazolyl tetrazolium (MTT) assay.Western blot was conducted to measure the protein expression of proliferating cell nuclear antigen (PCNA) and Notch-1 in HaCaT cells treated with recombinant TNF-α (25 ng/ml) and curcumin (20 μ mol/L) alone or in combination for 24 hours.Flow cytometry using annexin-V/propidium iodine (PI) was performed to assess the early apoptosis in HaCaT cells incubated with recombinant TNF-α of 25 ng/ml and curcumin of 20 μmol/L alone or in combination for 12 hours.Statistical analysis was carried out with one-way analysis of variance.Results Recombinant TNF-α promoted the proliferation of HaCaT cells in a dose-dependent manner,with the maximum proliferation activity observed in HaCaT cells treated with TNF-α of 25 ng/ml,while curcumin of 20 μmol/L effectively inhibited the proliferation of HaCaT cells induced by TNF-α of 25 ng/ml (P ＜ 0.01).TNF-α of 25 ng/ml had no obvious effect on cell apoptosis,while curcumin of 20 μ mol/L markedly induced the apoptosis in HaCaT cells,and there was a synergy between TNF-α of 25 ng/ml and curcumin of 20 μmol/L in the induction of apoptosis in HaCaT cells,with the apoptosis rate being 2.3％,3.4％,11.6％ and 16.8％ respectively in untreated cells,cells treated with TNF-α,curcumin,and the combination of TNF-α and curcumin,respectively.Conclusions Curcumin could enhance the inductive effect of TNF-α on the apoptosis in,but suppress the promotive effect of TNF-α on the proliferation of,HaCaT cells.

Objective To analyze the clinical characteristics of twin reversed arterial perfusion sequence (TRAP),and investigate its prenatal evaluation and clinical management.Methods Karyotype results and ultrasound data of 25 TRAP cases were retrospectively reviewed,including estimated weight and umbilical blood flow of acardiac twin,cardiac function and middle cerebral artery peak systolic velocity of pump twin.Various managements and the outcomes were analyzed.Results (1) Karyotype of amniotie fluid were tested in 16 pump twins.Mosaicism was found in 1 case (46,XX[36]/46,XY [14]).(2) According to the ultrasound evaluation,large acardia accounted for 87.0％ (20/23) cases.Abundant blood perfusion (inter-twin difference of umbilical resistance index ≤0.20) was indicated in 86.4％ (19/22) cases.Decompensation of cardiac function was suggested in 66.7％ (10/15) pump twins.Fetal anemia of pump twin indicated by middle cerebral artery peak systolic velocity＞1.5 multiples of the median was diagnosed in 75.0％ (12/16) cases.(3) The acardiac twin with abundant blood perfusion was more likely to be a large acardia than those without [94.7％(18/19) vs 1/3,Fisher exact test,P=0.04]; More pump twin with large acardia tended to have cardiac decompensation than non-large acardia pump twins [83.3 ％ (10/12) vs 0/3,Fisher exact test,P=0.02].(4) Eleven patients chose to terminate their pregnancies after being diagnosed.In 14 cases who continue the pregnancies,the survival rate of pump twin was 64.3％ (9/14).In 3 cases of non-large acardia without cardiac decompensation of pump twin,the patients selected conservative observation resulting in 2 term deliveries and 1 termination of pregnancy due to for exacerbation.Among 11 cases with large acardia,which the pump twins were complicated by cardiac decompensation or anemia,five cases selected conservative observation.One ended in spontaneous abortion; three exacerbated (one termination and two cesarean section before term with living births) ; one was stable until delivery.Another 6 cases received bipolar cord coagulation,and successful interruptions of acardiac blood flow were achieved in 5 cases among which 4 pump twins survived.Conclusions Prenatal diagnosis,cardiac function and fetal anemia of pump twin,together with the growth and blood supply of acardia are important indexes for prenatal evaluation of TRAP,on which our prompt management should be based.

Objective To evaluate the indications and intraoperative management of nephron-spar- ing surgery (NSS) for renal cell carcinoma (RCC).Methods A total of 53 patients (32 men and 21 women;age range,21 -76 years) with unilateral RCC and a normal contralateral kidney underwent NSS.The disease course ranged from 1 week to 13 months.The unilateral RCCs were detected by B-uhrasonography. Partial nephrectomy was performed on 38 cases of polar RCC and wedge nephrectomy was performed on 15 cases of central RCC.Both renal vein and artery were blocked for 10 - 15 min,and unclamped for 1 -2 min if necessary.The resections were finished within 7 -21 min (mean,12 min).No inosine and regional hypo- thermia were used during operation.Absorbable hemostatic gauze and terylene flake were used to suture renal wound surface.After operation all patients were treated with interferon hypodermic injection of 5 000 000 U every other day for 3 months.Results The diagnosis of RCC with negative cutting margin was confirmed by pathological examination in all patients.The greatest dimension of the renal cancers were＜4 cm except for one of 6 cm.The clinical stage was T_1N_0M_0 in all patients.During the follow-up of 6 -48 months,all pa- tients were alive without recurrence and had normal renal function.Conclusions NSS can be rapidly per- formed for T_1 stage renal cancer＜4 cm or single clearly localized cancer＞4 cm in diameter.The follow-up results are satisfactory.

Objective To summarize our preliminary experience of selective fetieide with bipolar coagulation in complicated monochorionie twins(MCT),and discuss the clinical application of feticide in discordant MCT.Methods Three MCT with one twin anomaly.in which 2 had severe twin-twin transfusion syndrome(TTTS),stage Ⅳ ,and 1 had acardiac twin,were identified in the second trimester of pregnancy.To terminate the abnormal twin and isolate the co-twin's circulation completely.selective feticide was performed by umbilical cord occlusion with bipolar coagulation under guidance of ultrasound and fetoscopy.After each invasive procedure,serial monitoring was performed,including procedural complications,Doppler of fetal middle cerebral artery and umbilical artery.Pregnancies were followed up every 2 weeks for fetal growth until delivery.After birth the placentas and the terminated fetuses were examined.Result Cord occlusion was successfully accomplished in all 3 targeted fetuses,at 21,22 and 24 weeks of gestation respectively.One case with TTTS was complicated with rupture of the membrane in the terminated fetus at the 7th day after the procedure.and a healthy baby was born at 32 weeks.The other case with TTTS delivered a boy by cesarean section at 38 weeks.The third case with TRAP is at 35 weeks of gestations and under regular follow-up.Monochorionicity was confirmed by placental examination after delivery.and the effects of bipolar coagulation were observed at the,cord of terminated fetuses.Conclusions Umbilical cord occlusion witll bipolar coagulation is an effective procedure for selective feticide in MCT with one twin anomaly.The outcome of normal fetus can be favorable.