人性激素结合球蛋白的真核表达、纯化鉴定及质控品应用分析

目的构建人性激素结合球蛋白(SHBG)的重组真核表达载体,获得高纯度的人SHBG重组蛋白。方法根据人SHBG基因序列,利用Primer Premier 6.0设计引物,扩增获得人SHBG基因;构建人SHBG-pCMV3H重组真核表达载体;将其转染至HEK-293F细胞,提取人SHBG重组蛋白,并用ConA柱亲和层析法纯化;采用人SHBG检测试剂盒对其蛋白浓度进行鉴定。结果人SHBG基因产物长度与预期一致,约为1200 bp;测序比对分析结果显示人SHBG基因成功插入pCMV3H载体;电泳结果显示在相对分子质量约为42×10^(3)处有一明显条带,与预期蛋白质位置一致;人SHBG检测试剂盒鉴定结果显示人SHBG重组蛋白作为质控品与西门子、安图生物、罗氏一致性良好;含钙离子缓冲液中人SHBG重组蛋白稳定性优于不含钙离子缓冲液。结论真核表达人SHBG重组蛋白能够作为SHBG检测试剂的质控品,且各厂家检测一致性良好;钙离子有利于提升人SHBG蛋白的稳定性。

应用iTRAQ和ELISA研究血清ApoB、FN1、SHBG与布加综合征的相关性

应用同位素相对标记与绝对定量将技术(iTRAQ)对布加综合征(Budd-Chiari Syndrome,BCS)患者以及对照组血清中蛋白质进行差异分析,筛选出差异蛋白,并对筛查的部分差异蛋白进行酶联免疫吸附法(ELISA)验证。方法:选取34例BCS患者与34例下肢静脉曲张患者血清,随机选取4例BCS患者血清和4例对照组静脉曲张患者血清,应用iTRAQ技术筛选出具有显著差异表达蛋白质,后对筛选出的血清人载脂蛋白B(ApoB)、纤维连接蛋白(FN1)和人性激素结合球蛋白(人性激素结合球蛋白)进行酶联免疫吸附法(ELISA)验证。结果:iTRAQ筛选出vWF、TIMP-1、FN1、SHBG、ApoB在两组间存在差异(p≤0.05);Elisa结果显示BCS组ApoB、FN1、SHBG含量均高于对照组,差异有统计学意义(P<0.05)。结论:ApoB、FN1、SHBG可能参与了布加综合征的病理过程。

Gastric choriocarcinoma admixed with an α-fetoprotein-producing adenocarcinoma and separated adenocarcinoma

We report a case of gastric choriocarcinoma admixed with an α-fetoprotein (AFP)-producing adeno-carcinoma. A 70-year-old man was hospitalized for gastric cancer that was detected during screening by esophagogastroduodenoscopy (EGD). Initial laboratory data showed the increased serum level of AFP and EGD revealed a 5-cm ulcerofungating mass in the greater curvature of the gastric antrum. The patient underwent radical subtotal gastrectomywith D2 lymph node dissection and Billroth gastrojejunostomy. Histopathological evaluation confirmed double primary gastric cancer: gastric choriocarcinoma admixed with an AFP-producing adenocarcinoma and separated adenocarcinoma. At 2 wk postoperatively, his human chorionic gonadotropin and AFP levels had reduced and six cycles of adjuvant chemotherapy were initiated. No recurrence or distant metastasis was observed at 4 years postoperatively.

Second-trimester maternal β-human chorionic gonadotropin level associated with subsequent development of pregnancy-induced hypertension

Objective:To determine whether maternal β-human chorionic gonadotropin(β-hCG) level in second-trimester may be associated with subsequent development of pregnancy-induced hypertension(PIH).Methods:Seven hundred and sixty-two women in mid-trimester were to have maternal urine β-hCG standardized concentrations and maternal serum β-hCG measurements.Their case histories were recorded and reviewed from mid-trimester to delivery.The relation was observed between maternal urine,serum markers and subsequent development of PIH.Results:Among 762 women,504 cases were normal pregnancies,42 cases had PIH,94 cases had premature rupture of membrane (PROM),69 cases had preterm delivery (PD),53 other cases were excluded by various reasons.The levels of maternal urine,serum β-hCG in PIH were (61.75±9.78) IU/L and (304.56±54.17) ng/mg respectively,which were higher significantly than normal pregnancy group ([20.65±7.61] IU/L and [146.34±47.81] ng/mg,P<0.05).When maternal serum,urine β-hCG levels ≥2 MOM(multiple of mean),the incidences of developing PIH were increased significantly as compared with those of β-hCG <2 MOM women.The incidence of PIH increased from 5.1% in pregnancies with urine β-hCG ≥2 MOM to 11.7% in cases with urine β-hCG ≥4 MOM.Conclusion:The elevation of maternal mid-trimester urine,serum β-hCG levels is not only an early signal for dysfunction of placenta but also a dangerous signal for development of PIH.Second-trimester maternal urine β-hCG measurement proves to be superior to serum marker in clinical prediction.

Recent Possibilities for the Diagnosis and Pharmacological Control of Pregnancy Loss in Dairy Cow

One of the most recent techniques for the diagnosis of EP （early pregnancy） in cattle on the farm is B-mode ultrasonography. Under field conditions, acceptable results may be achieved with ultrasonography from days 25 to 30 post-AI. The reliability of the test greatly depends on the frequency of the transducer used, the skill of the examiner, the criterion used for a positive PD （pregnancy diagnosis） and the position of the uterus in the pelvic inlet. Pregnancy protein assays （PAG, PSPB） may provide an alternative method to ultrasonography for determining EP or LEM/EFM （late embryonic/early foetal mortality） in the cow. Although early pregnancy factor is the earliest specific indicator for fertilization, its detection is entirely dependend on the use of the RIT （rosette inhibition test）, therefore its use in the field needs further inventions. Preventive pharmaceutical treatments with hCG （human chorionic gonadotropin）, GnRH （gonadotropin realising hormone）, PRID （progesterone-releasing intravaginal device） or CIDR （controlled internal drug release） inserts at different time periods at or post-AI may have some benefit in decreasing PL （pregnancy loss）, however further examinations are warranted to determine how and when these treatments may influence PL in the field.

Impact of the diameter of SonoVue microbubbles on binding characteristics of a new contrast agent targeted to choriocarcinoma cells in vitro

Objective:To evaluate the impact of the diameter of SonoVue microbubbles on binding characteristics, including the adhesion rate and stability, of a new contrast agent targeted to choriocarcinoma cells(JARs) in vitro, in order to establish a foundation to explore targeted ultrasound imaging for localization of tumor cell antigens and increase the early diagnostic rate for tumors.Methods:The objects were divided into three groups:the large microbubble group(n = 15), the middling microbubble group(n = 15) and the tiny microbubble group(n = 15).The rosette formation rate was counted.JARs were calculated by flow cytometry(FCM).The targeted contrast agent was prepared by mixing SonoVue microbubbles of different diam-eter with rabbit anti-human chorionic gonadotrophin(HCG) antibody.The binding rates of the targeted contrast agent to JARs before and after PBS rinse were analyzed.Results:The binding rate was significantly lower in the large microbubble group(61.7 ± 1.8)% than in the middling microbubble group(82.6 ± 4.5)% and the tiny microbubble group(91.3 ± 5.8)%(P < 0.05).The binding rates of different diameter microbubbles to JARs before and after PBS rinse were different.The middling microbubbles were the most stable ones, with the binding rate of(82.3 ± 4.5)% and(80.4 ± 3.9)% before and after PBS rinse(P > 0.05).The binding rates of the targeted microbubbles labeled with fluorescence to JARs were 68.6%, 81.3% and 89.3% in the large microbubble group, the middling microbubble group and the tiny microbubble group, respectively(P < 0.05).Conclu-sion:The binding capacity of the targeted SonoVue microbubbles to JARs is related to the diameter of the microbubble, which is determined by the shaking method before preparation.Modulating the diameter of SonoVue microbubbles may increase the binding rate and stability of targeted microbubbles to JARs, thus to improve the image of JARs.

The Roots of Psychological Trauma and Post-TraumaticStress Reaction/Disorders and the Intervention

Psychological trauma refers to the catastrophic or traumatic events harmful to individuals＇ inner world. Strong emotional and psychological reaction caused by traumatic factors will form psychological shadows, and the improper coping of these events will lead to a post-traumatic stress disorder. In this paper, the author makes an exploration on the roots of psychological trauma and of post-traumatic stress reaction or disorders, which were caused by natural traumatic event, such as floods, snowstorms, earthquakes and other man-made factors such as wars, accidents from a psychological point of view. Finally, some suggestions of intervention are provided for the relevant departments to make relevant rebuilding policies.

CHROMATOGRAPHIC SEPERATION AND PURIFICATION OF HUMAN CHORIONIC GONADOTROPIN

Seperation and purifitation of human chorionic gonadotropion (HCG) in theurine sample of early pregnant women by D3520 resin adsorption chromatographyis repaied. The crude product obtained by DEAE - Cellulose 23 and DEAE Sephadex A50 column chromatography showed a high activity of HCG. Furtherpurifikation of the sample by gel filtration chromatography on a Sephadex G75 column gives a final preparation of 6000～ 6500 IU/mg. The preparation meets therequiTements of the pyrogn test in Chinese Law of Pharmacopoeia.

Maximal androgen blockade versus castration alone in patients with metastatic prostate cancer

Maximum androgen blockade （MAB）, consisting of an antiandrogen plus either a luteinizing hormone- releasing hormone agonist （LHRHA） or orchiectomy, is a standard care for patients with prostate cancer. Although, clinical trial results have been equivocal, none has shown a significant advantage in favor of MAB over castration alone in metastatic prostate cancer and MAB has been the subject of considerable controversy. The aim of this study was to compare MAB （orchiectomy or LHRHA＂Goserelin＂） and anti-androgen ＂Bicalutamide＂ with castration alone （orchiectomy or LHRHA） in previ- ously untreated metastatic prostate cancer patients. Methods： Hundred eligible patients with adequate performance status and adequate hematologic, hepatic and renal functions were included. MAB arm, fifty patients underwent castration either surgically by orchiectomy or medically by receiving Goserelin （3.6 rag） depot, which was injected subcutaneously every 28 days plus bicalutamide 50 mg once daily. Castration alone arm, fifty patients underwent castration alone either surgically by orchiectomy or medically by receiving Goserelin （3.6 mg） depot. Results： During the period from January 2011 to January 2013, with a median follow up of 18 months （range 6 to 24 months）, there were eight deaths （16%）, in MAB arm and ten deaths （20%） in castration alone arm. At three months, there were 35 patients （70%） with prostate specific antigen （PSA） normalization （-〈 4 mg/dL） in MAB arm versus 17 patients （34%） with PSA normalization in castration alone arm （P = 0.001）. The median progression free survival （PFS） times were 22.18 months （95% CI, 19.7 to 24.2 months） for MAB arm versus 22 months in castration alone arm （95% CI, 18 to 25.9 months; P = 0.045）. The survival rates for MAB arm were 82% at 18 months and 70.6% at 24 months versus 78.7% at 18 months and 75.1% at 24 months in castration alone arm （P 〉 0.05）. The median overall survival （OS） was not reached in either arm. Both hematological and non-hematological toxicities were com- parable in both arms. Conclusion： MAB significantly improves the PSA normalization rate at 12 weeks and PFS compared to castration alone with no significant difference in overall survival and with comparable acceptable toxicities. However further studies are needed to document such findings.

多听音乐可预防

聆听或演唱自己喜欢的歌曲能使老年人性激素的分泌量趋于稳定，有助于预防阿尔茨海默氏症（老年性痴呆）。科学家在对36名老年妇女的检测中发现，音乐活动后，这些妇女的性激素分泌量整体向一个稳定值靠拢。在随后的心理测试中，这些妇女抑郁和不安的情绪也有所缓和。

别让疾病扫了你的“性”

随着年龄的增长,中老年人性激素的分泌逐渐减少,导致女性停经、男性精液和精子生成减少,使不少老年人停止了性活动。还有不可忽略的一点就是老年人常常患有各种慢性疾病。这些慢性病在很大程度上起了推波助澜的作用,它们使很多中老年人过早地远离了自己的“性”福。下面我们列举几种中老年人常见疾病对性的影响及应对方法。

长效β受体激动剂将再做安全性临床试验

为了进一步评价长效B受体激动剂（LABA）在治疗支气管哮喘中与吸人性激素联合应用的安全性，FDA要求LA_BA的生产厂家完成5项随机、双盲、对照临床试验，在治疗支气管哮喘中比较吸人性激素联合LABA与单纯应用吸人性激素的区别。目前有4项临床试验将在12岁以上的年轻人和成人中开展，每项临床试验将包含11700名患者，

Amomum villosum induces longitudinal bone growth in adolescent female rats

OBJECTIVE:This study was conducted to evaluate the effect of Amomum villosum on longitudinal bone growth.METHODS:Adolescent female Sprague-Dawley rats were divided into 3 groups and treated for 4 days:control(distilled water,p.o.),recombinant human growth hormone(rhGH;100 μg/kg,s.c.),and A.villosum(500 mg/kg,p.o.) groups.On day 3,tetracycline(20 g/kg,i.p.) was injected for growth plate identification.On days 2,3 and 4,5-bromo-2'-deoxyuridine(BrdU)(50 mg/kg,i.p.) was injected to label proliferating cells.On day 5,tibias were dissected and fixed in 4% paraformaldehyde,dehydrated,and sectioned for immunohistochemistry and histomorphometry.RESULTS:The rate of bone growth in the A.villosum and rhGH groups increased to(410 ± 44) and(389 ± 46) μm/day(P<0.01),respectively,as compared with the control(330.7 ± 34.7) μm/day.The thickness of the growth plates also increased to(591 ± 37) and(598 ± 32) μm,respectively,as compared with the control(524± 89) μm(P<0.001).The number of BrdU-positive cells in the chondrocytes of the A.villosum and rhGH groups was also significantly higher(126±24) and(143±18) cells/mm 2,respectively) than in the control(109 ± 25) mm 2(P<0.05).Insulin-like growth factor-1 and bone morphogenetic protein-2 in the A.villosum and rhGH groups were highly expressed in the growth plate as compared with the control samples,indicating increased bone formation.CONCLUSIONS:A.villosum could be used to treat growth retardation during adolescence.

Effect of soothing liver therapy on oocyte quality and growth differentiation factor-9 in patients undergoing in vitro fertilization and embryo transfer

OBJECTIVE:To investigate the effect of Soothing liver therapy on infertile women undergoing in vitro fertilization and embryo transfer(IVF-ET)and to explore its mechanism.METHODS:Fifty-eight women with tubal infertility were randomized into two groups:30 in an experimental group treated with Xiaoyao powder(Shugan)plus gonadotropin-releasing hormone analog(GnRHa)/follicle-stimulating hormone(FSH)/human chorionic gonadotropin(hCG)and 28 in the control group who were treated with GnRHa/FSH/hCG only.The total gonadotropin(Gn)doses required,endometrial thickness,oocyte numbers,high quality embryo production rate and pregnancy rate of the two groups were compared.The concentration of growth differentiation factor-9(GDF-9)in follicular fluid was detected by western blotting and the expression of GDF-9 mRNA in granulosa cells was measured using reverse tran-scription-polymerase chain reaction amplification.RESULTS:In the experimental group,the Gn dose was significantly lower than that in the control group;the endometrial thickness,high quality embryo production and pregnancy rates were significantly higher and the expression of GDF-9 mRNA was also significantly higher than in the control group(all P<0.05).CONCLUSION:Shugan treatment can improve the pregnancy rate of women with tubal infertility;its mechanism is possibly related to the increased expression of GDF-9 in granulosa cells.