Effect of Luteinizing Hormone-Releasing Hormone Analogue on the Sexual Behavior of Sacalia quadriocellata

Luteinizing hormone-releasing hormone(LHRH) is known to influence sexual behavior in many vertebrate taxa, but there have been no systematic studies on the role of LHRH in sexual behavior of turtles. We tested the hypotheses that exogenous LHRH analogues would induce sexual behavior of male Four-eyed turtle, Sacalia quadriocellata. We examined this by challenging males with intramuscular injections of mammalian luteinizing hormone-releasing hormone analogue(LHRH-A), human chorionic gonadotropin(HCG), or a combination of the two, and subsequently exposing them to sexually receptive females for behavioral observation. Our data show that the injection of only HCG could not, while that of only LHRH-A could, facilitate sexual behavior along with testicular recrudescence and spermatogenesis in S. quadriocellata. The injection of both LHRH-A and HCG would induce more drastic sexual behavior of the animals than that of LHRH-A alone, indicating HCG enhances the effects of LHRH-A induced sexual behavior. However, different pharmacological dosages of LHRH-A(0.5 μg, 1 μg, 2 μg per 100 g bodyweight) did not correspond to different activity levels. Though the mechanism of LHRH effect was not determined, this study may support that the sexual behavior of S. quadriocellata which occurs at the beginning of the injection despite regression of the gonads. This is the first report on the exogenous LHRH-A induced sexual behavior for this species.

Combination immunotherapy with Survivin and luteinizing hormone-releasing hormone fusion protein in murine breast cancer model

AIM To investigate the therapeutic potential of two recombinant proteins, Survivin and luteinizing hormone-releasing hormone (LHRH) fusion protein [LHRH(6 leu)-LTB] for immunotherapy of breast cancer.METHODS Murine 4 T-1 breast cancer model was used to evaluate the efficacy of recombinant proteins in vivo. Twenty four Balb/c mice were divided into 4 groups of 6 mice each. Recombinant Survivin and LHRH fusion protein, alone or in combination, were administered along with immunomodulator Mycobacterium indicus pranii (MIP) in Balb/c mice. Unimmunized or control group mice were administered with phosphate buffer saline. Each group was then challenged with syngeneic 4 T-1 cells to induce the growth of breast tumor. Tumor growth was monitored to evaluate the efficacy of immune-response in preventing the growth of cancer cells.RESULTS Preventive immunization with 20 μg recombinant Survivin and MIP was effective in suppressing growth of 4 T-1 mouse model of breast cancer (P = 0.04) but 50 μg dose was ineffective in suppressing tumor growth. However, combination of Survivin and LHRH fusion protein was more effective in suppressing tumor growth (P = 0.02) as well as metastasis in vivo in comparison to LHRH fusion protein as vaccine antigen alone.CONCLUSION Recombinant Survivin and MIP suppress tumor growth significantly. Combining LHRH fusion protein with Survivin and MIP enhances tumor suppressive effects marginally which provides evidence for recombinant Survivin and LHRH fusion protein as candidates for translating the combination cancer immunotherapy approaches.

Long-term effectiveness of luteinizing hormone-releasing hormone agonist or antiandrogen monotherapy in elderly men with localizect prostate cancer （T1-2） ： a retrospective study

Aim： To evaluate the long-term effectiveness, side effects and compliance rates of two types of drugs （luteinizing hormone-releasing hormone [LHRH] agonist and antiandrogen） that were used individually to treat patients with localized prostate cancer （T1-2） at our institution. Methods： Ninety-seven patients who were diagnosed in the period from April 1997 to January 2000 as having clinically localized prostate cancer （T1-2） received either LHRH agonist （leuprolide acetate 7.5 mg/month） monotherapy （group 1, n = 62） or antiandrogen monotherapy （group 2, n = 35; 18 received bicalutamide 50 mg q.d., 13 received nilutamide 150 mg t.i.d, and 4 received flutamide 250 mg t.i.d.）. The mean age in both groups was 76 years. Results： The mean follow-up time was （50.8 ±8.5） months in group 1 and （43.1 ± 2.2） months in group 2. Prostate-specific antigen （PSA） levels rose in only 1 of the 62 patients （1.6%） in group 1, and in 20 of the 35 patients （57.1%） in group 2. In group 2, 10 of the 20 patients （50 %） with increasing PSA levels were treated with LHRH salvage therapy, and eight （80%） responded. Hot flashes （54.8%） and lethargy （41.9%） were the most common side effects in group 1. In contrast, nipple-tenderness （40%） and light-dark adaptation （17.1%） were more often seen in group 2. Only 1 of the 62 patients （1.6%） in group 1 switched to another medication because of adverse side effects; whereas 8 of the 35 patients （22.9%） in group 2 did so. Conclusion： Unlike antiandrogen monotherapy, LHRH agonist monotherapy provided long-term durable control of localized prostate cancer （T1-2）. It can also be an effective treatment option for patients whose disease failed to respond to antiandrogen monotherapy. The limitations of our study are the lack of health outcomes analysis and a small sample size.

Preclinical therapy of benign prostatic hyperplasia with neuropeptide hormone antagonists

Benign prostatic hyperplasia(BPH)is a pathologic condition of the prostate described as a substantial increase in its number of epithelial and stromal cells.BPH may significantly reduce the quality of life due to the initiation of bladder outlet obstruction and lower urinary tract syndromes.Current medical therapies mostly consist of inhibitors of 5α-reductase orα1-adrenergic blockers;their efficacy is often insufficient.Antagonistic analogs of neuropeptide hormones are novel candidates for the management of BPH.At first,antagonists of luteinizing hormone-releasing hormone(LHRH)have been introduced to the therapy aimed to reduce serum testosterone levels.However,they have also been found to produce an inhibitory activity on local LHRH receptors in the prostate as well as impotence and other related side effects.Since then,several preclinical and clinical studies reported the favorable effects of LHRH antagonists in BPH.In contrast,antagonists of growth hormone-releasing hormone(GHRH)and gastrin-releasing peptide(GRP)have been tested only in preclinical settings and produce significant reduction in prostate size in experimental models of BPH.They act at least in part,by blocking the action of respective ligands produced locally on prostates through their respective receptors in the prostate,and by inhibition of autocrine insulin-like growth factors-Ⅰ/Ⅱand epidermal growth factor production.GHRH and LHRH antagonists were also tested in combination resulting in a cumulative effect that was greater than that of each alone.This article will review the numerous studies that demonstrate the beneficial effects of antagonistic analogs of LHRH,GHRH and GRP in BPH,as well as suggesting a potential role for somatostatin analogs in experimental therapies.

Real-world effectiveness and safety of goserelin 10.8-mg depot in Chinese patients with localized or locally advanced prostate cancer

Objective:Real-word data on long-acting luteinizing hormone-releasing hormone(LHRH)agonists in Chinese patients with prostate cancer are limited.This study aimed to determine the real-world effectiveness and safety of the LHRH agonist,goserelin,particularly the long-acting 10.8-mg depot formulation,and the follow-up patterns among Chinese prostate cancer patients.Methods:This was a multicenter,prospective,observational study in hormone treatment-na?ve patients with localized or locally advanced prostate cancer who were prescribed goserelin 10.8-mg depot every 12 weeks or 3.6-mg depot every 4 weeks with or without an anti-androgen.The patients had follow-up evaluations for 26 weeks.The primary outcome was the effectiveness of goserelin in reducing serum testosterone and prostate-specific antigen(PSA)levels.The secondary outcomes included testosterone and PSA levels,attainment of chemical castration(serum testosterone<50 ng/d L),and goserelin safety.The exploratory outcome was the monitoring pattern for serum testosterone and PSA.All analyses were descriptive.Results:Between September 2017 and December 2019,a total of 294 eligible patients received≥1 dose of goserelin;287 patients(97.6%)were treated with goserelin 10.8-mg depot.At week 24±2,the changes from baseline[standard deviation(95%confidence interval)]in serum testosterone(n=99)and PSA(n=131)were-401.0 ng/d L[308.4 ng/d L(-462.5,-339.5 ng/d L)]and-35.4 ng/m L[104.4 ng/m L(-53.5,-17.4 ng/m L)],respectively.Of 112 evaluable patients,100(90.2%)achieved a serum testosterone level<50 ng/d L.Treatment-emergent adverse events(TEAEs)and severe TEAEs occurred in 37.1%and 10.2%of patients,respectively.The mean testing frequency(standard deviation)was 1.6(1.5)for testosterone and 2.2(1.6)for PSA.Conclusions:Goserelin 10.8-mg depot effectively achieved and maintained castration and was well-tolerated in Chinese patients with localized and locally advanced prostate cancer.

野生鲇鱼生长激素分泌的季节变化及其神经内分泌调控

采用离体垂体碎片灌流孵育系统 ,将处于性腺退化期野生鲇鱼垂体切成约 1mm3 的碎片 ,用M 199冲洗之后放入灌流柱的两层Cytodex -Ⅲ微载体之间 (温度为 19± 1℃ )。每 5分钟收集一管灌流液 ,- 2 5℃贮存待测GH。采用鲤鱼GH放射免疫测定方法 (cGHRIA)测定鲇鱼垂体碎片灌流液以及血清和垂体中的GH含量。结果表明 :促黄体素释放激素类似物 [desGly10 (D Ala6)LHRHethylamide ,LHRH A]不能显著刺激离体垂体碎片基础GH分泌 ,注射LHRH A后不能显著提高血清基础GH水平 ;注射DA能显著提高鲇鱼血清基础GH水平 ,APO能以剂量依赖方式显著刺激垂体碎片基础GH分泌。雌、雄鲇鱼血清GH水平在 6月达到峰值 ,垂体GH水平在 3月和 7月份各出现一个峰值 ,各个季节雌鱼垂体和血清GH水平均显著高于雄鱼。鲇鱼血清和垂体GH水平与生殖周期有密切联系。

促黄体素释放激素类似物和多巴胺对鲤鱼幼鱼和性成熟雌鱼生长激素分泌的作用

以性腺未发育的５～６月龄幼鲤和１龄性成熟雌鲤为材料，研究年龄对促黄体素释放激素类似物和多巴胺能药物刺激的鲤鱼ＧＨ分泌的影响。结果表明：５～６月龄幼鱼血清基础ＧＨ水平明显低于１龄性成熟鱼；５～６月龄幼鱼对ＬＨＲＨＡ、ＤＡ和ＡＰＯ刺激的ＧＨ分泌的反应性也明显低于１龄性成熟鱼；ＤＡ和ＡＰＯ在５～６月龄幼鱼和１龄性成熟鱼都不能增强ＬＨＲＨＡ对ＧＨ分泌的刺激作用；ＤＯＭ不影响５～６月龄幼鱼和１龄性成熟鱼的血清ＧＨ水平。

性早熟女童血浆ghrelin水平与腺垂体激素的关系

目的：探讨性早熟女童血浆ghrelin与腺垂体激素的关系。方法：84例年龄在6-9岁的性早熟女童根据第二性征发育情况、骨龄、子宫、卵巢容积及GnRH激发试验的结果分为特发性中枢性早熟（ICPP）和单纯乳房早发育（PT）组,采用放射免疫法测定性早熟女童空腹血浆ghrelin水平,化学发光法测定ACTH、TSH、PRL、GH、LH、FSH,并与对照组比较。结果：1ICPP组血浆ghrelin为Log（2.42±0.26）ng/L,明显低于PT组和对照组[Log（2.62±0.21）ng/L,Log（2.58±0.44）ng/L,P〈0.05];而对照组与PT组两组之间比较差异无显著性（P〉0.05）。2TannerⅢ期的ICPP女童ghrelin为Log（2.31±0.24）ng/L明显低于TannerⅡ期的ICPP女童[Log（2.53±0.24）ng/L,P〈0.05]。3经双变量相关分析,性早熟女童血浆ghrelin水平与ACTH、PRL及GnRH激发后的LH15、LH30、LH60呈负相关（r分别为-0.248、-0.235、-0.445、0.405、0.398,均P〈0.05）;而与GH、GnRH激发后的FSH15及LH0^-2、FSH0^-2无相关。结论：ICPP女童血浆ghrelin水平下降,TannerⅢ期女童血浆ghrelin低于TannerⅡ期,GnRH激发后与LH呈负相关。这可能与性启动有关。

寡肽(LRH-A)分子量及一级结构的基质辅助红外激光解吸质谱分析

应用基质辅助红外激光解吸离子化/博里叶变换质谱法(IR-MALDI/FTMS),对九肽药物促黄体素释放激素类似物(LRH-A)的分子量及其氨基酸序列进行测定。用2,5-二羟基苯甲酸(DHB)作为基质,测得准分子离子(M+H)~1的m/z=1167.61006,与理论计算值的相对误差为3.66ppm。调节激光能量,获得一系列典型的碎片离子,从而确证了LRH-A的氨基酸序列。此法不需对样品进行预处理。

药物性卵巢去势治疗绝经前复发乳腺癌临床疗效观察

目的观察黄体生成素释放激素(luteinizinghormonereleasinghormone,LH-RH)类似物治疗绝经前复发性乳腺癌的效果及安全性。方法对28例绝经前复发乳腺癌患者进行LH-RH类似物治疗,观察疗效与不良反应。结果治疗后完全缓解+部分缓解(CR+PR)12例,有效率为42.9%,治疗8周后血浆雌激素降至绝经后水平。结论LH-RH类似物治疗绝经前乳腺癌疗效肯定,副作用小,给药方便。

电喷雾质谱法(ESI-MS)分析两种寡肽的一级结构

目的:测定肽类药物曲普瑞林、促黄体素释放激素类似物的分子量及其一级结构。方法:应用电喷雾质谱法(ESI MS)及源内碰撞诱导解离技术对肽类药物曲普瑞林及促黄体素释放激素类似物进行测定,根据其典型碎片离子确证其氨基酸序列及分子量。结果:肽类化合物在正离子模式下可得到较好的质谱信息,2种寡肽均测得其准分子离子峰(M+H)^+及(M+Na)^+信号,分子量分别与理论值一致。利用电喷雾碰撞诱导解离方法,得到一系列典型的 y 及 b 系列碎片离子,从而确证了2种寡肽的一级结构。结论:本研究为测定寡肽的分子量和一级结构提供了一个简便、快速、准确的方法。

国产黄体生成素释放激素─A治疗子宫内膜异位症的临床研究

应用不同剂量（１００μｇ／ｄ，２００μｇ／ｄ）的国产黄体生成素释放激素－Ａ（ＬＨＲＨ－Ａ）治疗子宫内膜异位症６６例，其中Ⅰ期９例，Ⅱ期４７例，Ⅲ期１０例。合并不孕症２９例，子宫肌腺症１０例。结果表明，５００μｇ／ｄ组对卵巢内膜囊肿的疗效最佳，但对子宫腺肌症的效果不明显，停药后妊娠７例，占２４％（７／２９）。所用ＬＨＲＨ－Ａ的３种剂量均抑制Ｅ２，Ｐ，ＰＲＬ的分泌，使血中生殖激素下降，排卵抑制和卵巢内膜囊肿缩小，对血胆固醇有升高作用，临床主客观症状体征以５００μｇ／ｄ组变化最为明显。

LHRH固体脂质纳米粒口服给药系统的研究

目的考察以固体脂质纳米粒为载体包裹促黄体生成素释放激素的类似物(LHRH-A2)口服给药的可行性。方法采用溶剂扩散法制备LHRH-A2单硬脂酸甘油酯纳米粒(LHRH—A2 loaded monostearin solid lipid nanoparticles,LHRH-A2-MSLN),考察载药纳米粒的粒径、Zeta电位等理化性质;用HPLC测定药物的包封率及体外释放特性;以花(鱼骨)鱼(Hemibarbus maculates Bleeker)为模型动物,载药纳米粒经口灌给药后,分别测定鱼的受精率和催产率。结果LHRH-A2-MSLN的粒径为(284.1±103.6)nm,纳米粒Zeta电位为-(19.4±1.1)mV,HPLC测得药物包封率为63.4%。体外释放研究结果显示,LHRH-A2-MSLN在前8 h快速释放了纳米粒所载药物的26.8%,随后每天以1.9%的速率持续释放。体内药效学研究表明,50μg·kg-1剂量的LHRH-A2-MSLN 口灌给药后,平均催产率46.5%,受精率90.5%。结论用溶剂扩散法制备得到的LHRH-A2-MSLN具有一定的口服有效性。

基质辅助红外激光解吸质谱法在寡肽分析中的应用

用基质辅助红外激光解吸离子化／傅里叶变换离子回旋共振质谱法（ＩＲＭＡＬＤＩ／ＦＴＩＣＲＭＳ）分析了促黄体素释放激素类似物、血管紧张肽原酶底物及血管紧张肽。分别以２，５二羟基苯甲酸、丁二酸为基质，测定其分子量，测定值与理论计算值的相对误差小于４×１０－６，适当增加激光功率密度，获得了一系列特征碎片离子并进行了归属，从而确证了３种寡肽的一级结构。提示本研究为测定寡肽的分子量和一级结构提供了一个简便、快速。

Targeted therapy in advanced metastatic colorectal cancer: Current concepts and perspectives

The introduction of new cytotoxic substances as well as agents that target vascular endothelial growth factor (VEGF) and epidermal growth factor receptor (EGFR) signaling has improved clinical outcome of patients with metastatic colorectal cancer (mCRC). In this review we summarize the most relevant clinical data on VEGF and EGFR targeting regimens in mCRC. The effects of available treatment strategies for mCRC are often temporary, with resistance and disease progression developing in most patients. Thus, new treatment strategies are urgently needed. Some GI peptides including gastrin and gastrin releasing peptide, certain growth factors such as insulin-like growth factor-I&#x02005;and II and neuropeptides such as growth hormone releasing hormone (GHRH) are implicated in the growth of CRC. Experimental investigations in CRC with antagonistic analogs of bombesin/gastrin-releasing peptide, GHRH, and with cytotoxic peptides that can be targeted to peptide receptors on tumors, are summarized in the second part of the review.

Hormone naive prostate cancer： predicting and maximizing response intervals

Hormone naive advanced prostate cancer is subdivided into two disease states： biochemical recurrence and traditional M 1 （metastatic） prostate cancer and characterized by no prior hormonal therapy or androgen deprivation therapy （ADT）. In biochemical recurrence/ prostate-specific antigen （PSA） recurrence, men should be risk-stratified based on their PSA doubling time, the Gleason score and the timing of the recurrence. In general, only men who are at high risk should be considered for early/immediate ADT although this is best done using shared decision with the patient. The type of ADT to be used in biochemical recurrence ranging from oral-only peripheral blockade （peripheral androgen deprivation） to complete hormonal therapy （combined androgen blockade [CAB]） remains in debate owing to lack of randomized controlled trials （RCT）. However, there is good RCT support for use of intermittent hormonal therapy （IHT）. There is also limited research on biomarker response （PSA and testosterone decline） to predict prognosis. On the other hand, in the setting of M1 hormone naive prostate cancer, there are many more RCT＇s to inform our decisions. CAB and gonadotrophin-releasing hormone antagonists perhaps provide a slight efficacy advantage while IHT may be slightly inferior with minimal M1 disease. The PSA nadir at 7 months after starting ADT is a powerful prognostic tool for M1 patients. There is growing recognition that serum testosterone （T） control while on ADT is linked to the development of castrate-resistant prostate cancer. Especially for a M 1 patient, maintaining a serum T below 20-30 ng d1-1 prolongs the response to ADT. Novel oral agents （abiraterone and enzalutamide） may soon find use in hormone naive disease and may alter the treatment landscape. Despite over 75 years of experience with ADT, many questions remain, and the field continues to evolve.

Regulative Actions of the Chinese Drugs for Tonifying the Kidney on Gene Expression of the Hypothalamic GnRH,Pituitary FSH,LH and Osteoblastic BGP

It is found that the drugs for nourishing yin to reduce pathogenic fire can significantly down-regulate,and the drugs for tonifying the kidney to replenish essence can up-regulate mRNA expression of the hypothalamic GnRH,pituitary FSH,LH and osteoblastic BGP,indicating that the Chinese drugs for tonifying the kidney can regulate gene expression of the hypothalamic GnRH,pituitary FSH,LH,and osteoblastic BGP,which is possibly one of the main mechanisms of the Chinese drug for tonifying the kidney,regulating ephebic development process and improving skeletal development in sexual precocity children.

注射大剂量促黄体素释放激素类似物后受孕大鼠黄体细胞的超微结构变化

受孕大鼠共20只,于受孕第9天随机取10只,注射大剂量促黄体素释放激素9肽类似物(LRH—A)后1和2天取材,作为给药组;其余10只为对照组。用透射电镜观察,比较两组大鼠黄体细胞的超微结构。给药组黄体细胞胞质内脂滴增多,出现许多大脂滴;滑面内质网排列紊乱、减少和溶解;线粒体有退化;自噬泡、溶酶体、髓样小体、多泡体以及残余体等结构增多;高尔基复合体在给药后1天肥大,以后转入萎缩。细胞表面微绒毛减少。以上结果提示,受孕9天大鼠注射大剂量 LRH-A后,黄体细胞在形态上趋向退化,在功能上分泌降低。

Kinetics of testosterone recovery in clinically localized prostate cancer patients treated.with radical prostatectomy and subsequent short-term adjuvant androgen deprivation therapy

deprivation therapy （ADT） is a standard treatment for metastatic, recurrent and locally advanced prostate cancer （PCa）. The aim of this study is to investigate the timing and extent of testosterone recovery in clinically localized PCa patients treated with radical prostatectomy （RP） and subsequent short-term adjuvant ADT. A total of 95 localized PCa patients underwent RP and 9-month adjuvant ADT were included in this prospective study. Serum testosterone level was measured before adjuvant ADT, at ADT cessation, and at 1, 3, 6, 9 and 12 months after cessation of ADT. A Cox proportional hazards model was used to assess variables associated with the ti me of testosterone normalization. The results showed that median patient age was 67 years and median testosterone level before adjuvant ADT was 361 （230-905） ng d1-1. All patients finished 9-month adjuvant ADT and achieved castrate testosterone level. At 3 months after ADT cessation, testosterone recovered to supracastrate level in 97.9% patients and to normal level in 36.9% patients. The percentage of patients who recovered to normal testosterone level increased to 66.3%, 86.3% and 92.6% at 6, 9 and 12 months, respectively. Cox regression model found that higher baseline testosterone level （ 300 ng dl- 1） was the only variable associated with a shorter time to testosterone normalization （hazard ratio： 1.98; P -- 0.012）. In conclusion, in most patients, testosterone recovered to supracastrate level at 3 months and to normal level at 12 months after 9-month adjuvant ADT cessation. Patients with higher baseline testosterone level need shorter time of testosterone normalization.