PCR 检测产 A、B、E、F 和 G 型肉毒神经毒素梭菌的神经毒素基因

肉毒神经毒素分为Ａ、Ｂ、Ｃ、Ｄ、Ｅ、Ｆ和Ｇ七个血清型，其中Ａ、Ｂ、Ｅ和Ｆ型引起人类肉毒中毒。为此，选取肉毒神经毒素的保守序列设计了一对简并引物，对１８株肉毒梭菌和８株相关梭菌进行了检测，表明该引物可特异扩增Ａ、Ｂ、Ｅ、Ｆ和Ｇ型肉毒梭菌及产Ｅ型肉毒神经毒素的丁酸梭菌，产物为２６４ｂｐ，敏感性达１０ｐｇ细菌ＤＮＡ。采有酚－氯仿抽提法、固相载体捕获法和热裂解法处理产毒菌株，结果表明热裂解法安全、简便、快速，所制模板扩增效果明显。因此，此ＰＣＲ方法可用于快速敏感地检测引起人类肉毒中毒的梭菌。     

Idiopathic anaphylaxis: long-term follow-up, cost, and outlook

To determine the efficacy of oral corticosteroids, antihistamines, and sympathomimetics in treating patients with idiopathic anaphylaxis, the charts of 225 patients diagnosed with idiopathic anaphylaxis from 1971 to 1990 treated at a single center were reviewed. Sixty-one patients (34 females and 27 males) were available for long-term follow-up. Ages ranged from 10 to 68 years with an average of 39 years. Patients with frequent episodes were treated with a protocol of oral corticosteroids, antihistamines, and sympathomimetics. Patients with infrequent episodes were treated for acute episodes only. The number of emergency room visits, hospitalizations, intensive care unit admissions, and length of time in remission were recorded. Sixty-five percent of patients with infrequent episodes and 91% of patients with frequent episodes of idiopathic anaphylaxis went into remission. Significant decreases in emergency room visits occurred for the idiopathic anaphylaxis-generalized-frequent group ( P< 0.016). the idiopathic-anaphylaxis-generalized-infrequent group ( P< 0.0001), and the idiopathic anaphylaxis-angioedema-infrequent group ( P <0.039). Significant decreases in the number of hospitalizations ( P < 0.022) and intensive care unit admissions ( P < 0.009) occurred for the idiopathic anaphylaxis-generalized-infrequent and frequent groups, respectively. Overall, an estimated 184740 was saved with the treatment program, for 546 patient-years. Idiopathic anaphylaxis can be controlled and remission induced in most patients. An estimated 11 million per year can be saved for patients in the USA on the basis of the estimated prevalence in this country.     

Acute,massive, haemorrhagic adrenal necrosis experimentally produced by the Shwartzman mechanism in rabbits

Summary Acute and severe haemorrhagic necrosis of the adrenal was produced experimentally in rabbits by means of intravenous injection of endotoxin after pretreatment by adrenocorticotropic hormone (ACTH) administration. The change occurred mainly in thezona fasciculata of the adrenal cortex, and its pathology was quite similar to that of the Shwartzman reaction. Numerous microthrombi were found in and around the lesion, but no marked changes were seen in other parts of the body. Heparin administration was very effective in preventing the necrosis. The pathogenesis of this lesion was postulated to be a univisceral Shwartzman mechanism in the adrenal. This seems to be a good experimental model for massive haemorrhagic necrosis of the adrenal in man, for example in the Waterhouse-Friderichsen syndrome, the pathogenesis of which has been assumed to involve intravascular clotting. It is suggested that hyperfunction of the adrenal cortex caused by ACTH administration could be a preparative condition for the Shwartzman reaction.This study was supported in part by a grant-in-aid from the Japanese Ministry of Education, Science and Culture     

快速AS-PCR技术用于血小板HPA-1，-2，-3，-4，-5抗原系统等位基因的分型

目的　研究建立快速等位基因特异性 (AS)引物 PCR技术 ,同时对人血小板同种异型抗原系统 (HPA 1,2 ,3,4,5 )等位基因进行分型。方法　设计合成 15条等位基因特异性引物 ,摸索最适引物浓度、Mg 浓度及扩增参数。用该技术对 10 0名北京地区健康献血者HPA 1～ 5系统等位基因进行分型。结果　从 10 0名北京地区献血员观察到的HPA基因频率分别是HPA 1a和 1b为 0 .995和 0 .0 0 5 ,HPA 2a和 2b为 0 .90 0和 0 .10 0 ,HPA 3a和 3b为 0 .775和 0 .2 2 5 ,HPA 4a和 4b为 1.0 0 0和 0 ,HPA 5a和 5b为 0 .970和 0 .0 30。结论　该方法简单、快速 ,结果准确 ,适用于临床及常规献血员血小板分型。     

中国广州汉族人群六个STR位点的调查

目的 了解广州汉族人在ｖＷＡ３１Ａ等６个短串联重复序列（ｓｈｏｒｔ ｔａｎｄｅｍ ｒｅｐｅａｔ,ＳＴＲ）位点法医学的有关数据。方法 运用ＳＴＲ－ＰＣＲ、４％变性聚丙烯酰胺凝胶电池,结合荧光ＤＮＡ自动测技术,对汉族人群６个ＳＴＲ位点的等位基因频率和基因型频率进行了调查,并与其他种族或人群的等位基因频率进行了比较。结果 ６个ＷＴＲ位点是ｖＷＡ３１Ａ、ＴＨ０１、Ｆ１２Ａ０１、Ｆ１３Ａ０１、ＦＥＳ、ＴＰＯ     

Effects of local treatment with salmeterol and terbutaline on anti-IgE-induced wheal, flare, and late induration in human skin

The capacity of terbutaline and the long-acting β₂-agonist salmetcrol to suppress wheal-and-flare reactions (WFRs) to intradermal antihuman IgE and to histamine was evaluated in 36 healthy volunteers. We also exaniined effects of the two drugs on the subsequent late cutaneous reaction (LCR) to anti-IgE. Salmeterol (10^-10 10^-6 M) and terbutaline (10^-10 10^-5 M), injected intradermally 2 or 15 min before anti-IgE challenge, produced a dose-dependent inhibition of the WFRs to anti-IgE (liter 1:10000) with a maximal effect of ?60% (wheal) and ??75% (flare) by both drugs. On a molar basis, salmctcrol was approximately 10–100 times more potent than terbutaline in inhibiting the WFRs. Moreover, the wheal response to histamine (4 nmol) was antagonized by ?40% after prctrcatment with either salmeterol (10^-5 M) or terbutaline (10^-4 M). We found that both salmeterol and terbutaline exhibited anti-WFR activity for up to 24 h, salmeterol being significantly more potent in this regard. When selecting a 15-min pretreatment interval with equieffective anti-WFR doses from the first dose-response experiments (i.e., a salmetcrol: terbutaline ratio of 1:10), the WFRs to high-dose anti-IgE (titer 1:100) were inhibited by terbutaline (10^-5 M) by 25–30%, but not by salmeterol (10^-4). On the other hand. salmeterol attenuated (by up to ?35%) the subsequent LCR more effectively than terbutaline. As compared to the prctrcatment procedure, infiltration of the drugs (single doses) into the wheals 30 min after challenge with anti-IgE (titer 1:100) proved to be less effective on the development of LCRs. Taken together, salmeterol was found to express higher potency and have longer duration of action than terbutaline in inhibition of IgE-mediated inflammation in human skin.     

Association of viral factors with non-familial breast cancer in Taiwan by comparison with non-cancerous, fibroadenoma, and thyroid tumor tissues

To study the etiologic factors of non-familial breast cancer, the polymerase chain reaction (PCR) and Southern hybridization were used to detect six viruses including human papillomavirus (HPV), cytomegalovirus (CMV), Epstein-Barr virus (EBV), herpes simplex virus (HSV)-1, HSV-2, and human herpesvirus (HHV)-8 DNA in 69 patients with breast cancer and 60 specimens from non-cancerous or other individuals with thyroid tumors or fibroadenoma (non-breast cancer controls). Two specimens from patients with a familial history of breast cancer and five breast cancer specimens with negative results for beta-globin, which was used as internal control, were excluded from this study. Eight (12.9%) HSV-1, 28 (45.2%) EBV, 47 (75.8%) CMV, 8 (12.9%) HPV, and 28 (45.2%) HHV-8 positive samples out of the 62 breast cancer specimens were detected; no HSV-2 DNA was detected in any group. Among the viral gene-positive breast cancer samples, 12 (23.1%) were positive for 1 virus, 16 (30.8%) were positive for 2 viruses, 21 (40.4%) were positive for 3 viruses, and 3 (5.8%) were positive for 4 viruses. Among the viral gene-positive specimens of the control groups, only one virus, CMV, was found in the non-cancerous and thyroid tumor specimens, while multiple viruses were found in the fibroadenoma specimens. The viruses associated with breast cancer were HHV-8 > EBV (P <0.01). The viruses associated with fibroadenoma were HSV-1 and HHV-8 > EBV (P <0.01). The presence of more than one virus was found predominantly in breast cancer and exclusively found in fibroadenoma. CMV was the only virus associated with thyroid tumors.     

Viral detection in hydrops fetalis, spontaneous abortion, and unexplained fetal death in utero

This study was undertaken to investigate the occurrence of viral infection in fetal death by examining tissues for the presence of DNA of several viral agents. Tissue specimens including heart, kidney, liver, lung, and placenta of 73 cases of fetal death were examined with 27 cases of elective termination of pregnancy as a control group. DNA extracted from these samples was tested for the presence of HSV, CMV, EBV, VZV, HHV-6, HHV-7, and PVB19. Viral DNA was found in one or more tissue samples from 25/73 cases (34%): CMV in 20, HSV in 5, parvovirus B19 in 5, HHV-7 in 3, and HHV-6 in 2. The presence of HHV-6 in fetal tissue has been reported rarely. No study so far has reported the detection of HHV-7 in fetal tissues with normal or adverse outcomes. Viral DNA was not found in any of the termination of pregnancy samples. Among the positive cases, eight had dual infection. One further case was positive for three viruses: HSV, CMV, and HHV-7. HHV-6 was the sole infectious agent in two cases, HHV-7 in one case, PVB19 in three, and CMV in ten cases. The finding of multiple viral DNA in 12% of the cases suggests the involvement of complex risk factors in cases of fetal loss. Although the cause of fetal death often includes other factors (e.g., chromosomal abnormalities) these data suggest the incidence of viral infective etiology may be higher than considered previously. However, larger studies are required to establish this link.     

Inhibition of anti-IgE induced skin responsein normals by formoterol, a new β2-adrenoceptor agonist, and terbutaline

Formoterol, a new beta 2-selective long-acting bronchodilator, was compared with terbutaline in terms of ability to inhibit dual phase skin reactions to anti-human IgE in volunteers. Anti-IgE induced an early wheal and flare reaction (WFR) followed by a progressively increasing induration, the late phase reaction (LCR), lasting greater than or equal to 24 h. Intradermal injection of formoterol 20 ng or terbutaline 500 ng 5 min before challenge gave equal inhibition of the WFR. The subsequent LCR was suppressed by formoterol (30%) for the whole 24 h period, while terbutaline only attenuated the first 4 h period. Increasing the dose range of both drugs 25-fold, caused a further analogous reduction of the WFR to anti-IgE. In this higher dose range formoterol (0.5 micrograms) antagonized the following 1-24 h LCR by 50%, while terbutaline (25 micrograms) only attenuated the LCR by an average of 20%, with higher effect in the first 6 h period. The anti-LCR capacity of formoterol was highly superior to that of terbutaline (P less than 0.001). The histamine-elicited wheal response was attenuated by both drugs, but they had no effect on the flare response, favouring an anti-permeability action of both compounds. The data support the concept that terbutaline, given locally in a single dose shortly before challenge, inhibits the mast cell mediator release reaction with limited consequences for the following LCR. In contrast to terbutaline, formoterol exerted a substantial anti-LCR action, probably by interfering with inflammatory mechanisms after the initial mast cell mediator release.     

Photoinduced Side-Chain Functionalization,Grafting, Cross-Linking,and Chain Extension by Thiol-Epoxide Click Reaction Using Photo Base Generator

In this work, photoinduced thiol-epoxy click reaction for side-chain functionalization, chain-extension, grafting, and cross-linking of polymers prepared by atom transfer radical polymerization by using 1,5-diazabicyclo[5.4.0]undec-5-ene thioxanthone acetate (Tx-DBU) as a photo base generator (PBG) is described. Irradiation of Tx-DBU at 365 nm leads to the generation of 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) in neutral form as a super base. DBU is capable of abstracting protons from thiols and thus formed thiolates are excellent nucleophiles that exhibit high affinity toward electrophiles such as epoxide rings. Appropriately selected thiols are successfully used for the desired reactions. The precursors and resulting polymers are characterized using ¹H-NMR, Fourier transform infrared, gel permeation chromatography, and differential scanning calorimetry measurements. This study shows that the use PBGs can further be extended to the synthesis of various complex macromolecular structures by the described post modification processes.     

HCV 5′ NCR和结构蛋白编码序列的拼接及在pLNSX中的克隆

目的:构建丙型肝炎病毒(HCV)5′末端非编码区(5′NCR)和结构蛋白编码基因序列逆转录病毒重组体,用于探索控制HCV感染的新途径和基因治疗。方法:对多株HCV核酸序列进行同源性比较设计引物,逆转录聚合酶链式反应(RT-PCR)扩增5′NCR、C、E1和E2/NS14个编码区共5个片段,分别克隆。以连接聚合酶链反应(PCR)将这5个片段拼接为一连续的长2547nt的片段,含HCV完整的5′NCR和全部的结构蛋白编码序列。将此序列插入pGEM-Zf(+)载体,与逆转病毒pLNSX载体中,转化大肠杆菌DH5a、转化菌落经酶切、PCR和Southern杂交鉴定。结果:通过RT-PCR和连接聚合酶链式反应(PCR)扩增出2547nt含HCV完整的5′NCR和全部的结构蛋白编码序列,将此序列与pGEM-Zf(+)重组得重组体pHC2547,与逆转录病毒载体pLNSX重组得pL-HC。结论:成功构建了HCV逆转病毒重组体,以利于HCV的胞内基因表达调控研究,更为探索HCV分子致病机制和转基因动物及基因治疗提供可靠的物质基础。     

Immunohistochemical,molecular, and pathomorphological study of liver biopsy specimens during chronic hepatitis C

We carried out a comparative study of replication markers of hepatitis C virus (HCV RNA) in some biological substrates and NS3-antigen in liver biopsy specimens. It was found that 82% liver specimens contained RNA HCV, in 44% cases HCV RNA was present in the serum, mononuclear blood cells, and liver. The presence of NS3-antigen in hepatocytes can be considered as a structural marker of HCV replication, which is confirmed by positive correlation with the results of PCR for viral RNA in tissue specimens. In most cases the presence of HCV replication markers did not correlate with activity of the infectious process assessed by biochemical and histological tests.     

Human papillomavirus 6, 11, and 16 in laryngeal papillomas.

Twenty-seven cases of benign laryngeal papillomas, both single and multiple variants, were analysed for human papillomavirus (HPV) by DNA slot-blot hybridization chiefly to determine the pattern of infection in Hong Kong Chinese. DNA was extracted from paraffin blocks of formalin-fixed tissue and probed separately for HPV 6, 11, 16, and 18. Sixteen cases (59 per cent) showed the presence of at least one of these four HPV genomes. Thirteen cases (48 per cent) were positive for HPV 11 only. Three other cases (11 per cent) showed triple positivity for HPV 6, 11, and 16. None were positive for HPV 18. The predominance of HPV 11 infection contrasts with other series which have shown either an almost equal distribution of HPV 6 and 11 or a predominance of HPV 6. The finding of HPV 16 in three cases was unexpected. Using the polymerase chain reaction (PCR) with primers complementary to the upstream regulatory region of the HPV 16 viral DNA, the presence of HPV 16 genome was confirmed in all three cases. As the number of HPV 16-positive cases in this study is small, analysis of more cases using fresh biopsy material and a wider range of HPV type-specific PCR primers is warranted to determine the relative incidence of HPV subtypes in these benign laryngeal papillomas.     

8例46,XX男性和16例46,XY女性的SRY序列研究

为探讨Ｙ染色体性别决定区基因（ＳＲＹ）在性分化中的作用。在染色体核型分析的基础上，应用聚合酶链反应（ＰＣＲ）和直接测序技术，对８名４６，ＸＸ男性和１６名４６，ＸＹ女性患者进一步作了ＳＲＹ序列的分析研究。结果发现：２例４６，ＸＸ男性的ＤＮＡＰＣＲ扩增出现特异的ＳＲＹ序列扩增片段；１例４６，ＸＹ女性的ＳＲＹ核心序列ｃｏｄｏｎ１１３出现Ａ→Ｔ的新生突变；１例４６，ＸＹ女性的ＳＲＹ编码序列上游ＡＡＣＡＡＧ区（转录因子结合位点）处碱基Ｇ→Ａ突变。上述研究结果提示：ＳＲＹ基因是性发育的重要遗传学基础，并为进一步揭示ＳＲＹ基因的结构与功能的关系和在临床上对性反转综合征患者的诊断、治疗提供有价值的科学资料。     

Development of multiplex real-time quantitative PCR for simultaneous detection of Chlamydia trachomatis,Mycoplasma hominis,Ureaplasma urealyticum,and Mycoplasma genitalium in infertile women

PurposeSexually Transmitted Diseases (STDs) can cause sterility and many other problems for women planning pregnancy. Currently, almost 340 million people worldwide suffer from Sexually Transmitted Infections (STIs). This study made attempts to quickly identify STDs' most critical infectious agents using dedicated primers and probes.MethodsThe present study was done on the cervical samples of 200 infertile women. After extracting the total DNA of Chlamydia trachomatis, Mycoplasma hominis, Ureaplasma urealyticum, and Mycoplasma genitalium, quantitative methods were employed to determine the rate of target bacteria using multiplex real-time PCR.ResultsThe multiplex qPCR showed the rates of 47%, 16%, 46%, and 16.5% for Chlamydia trachomatis, Mycoplasma hominis, Ureaplasma urealyticum, and Mycoplasma genitalium in infertile women, respectively. In some patients, there were co-infections with two or three bacteria. The diagnostic approach used in our research could be employed as an alternative detection tool to identify the four most common STD-associated bacterial agents while detecting mixed infections.ConclusionsInfertile women with no biological problems could have their genital tract checked using this newly designed identification technique and get proper treatment for their infections as quickly as possible.     

Molecular Identification and Detection of Virulent Factors in Helicobacter pylori from Gastric Biopsy Samples of Patients Attended at Assam Medical College and Hospital,Dibrugarh, Assam,India

The present study aimed to detect the major virulence determinants of Helicobacter pylori, the gastric bacteria by polymerase chain reaction from genomic DNA of 314 gastric biopsies from dyspeptic patients in 2015-2016 through upper gastrointestinal endoscopy. In 153 cases out of 314, the high prevalence of oipA gene followed by cagA-vacA s1 m1 combined genotypes was found in mostly gastritis/duodenitis patients followed by the peptic ulcer and normal patients. Therefore, the clinical significance of the virulence markers of H. pylori associated with the severe forms of gastroduodenal diseases is still a matter of controversy since the endoscopically normal patients were found to harbour the virulent genes.     

运动前后运动神经传导速度与体温、心率变化的相关分析

运动对神经传导速度的影响尚不十分清楚。本文观察了运动前后青年人胫神经MCV、体温、心率的变化,结果表明,一定运动后平均口温升高0.45℃,平均胫神经MCV增快5.11m/s,这与环境温度、体表温度对神经传导速度的影响有明显差异。应用多元回归方程分析发现,胫神经MCV变化与体内温度变化相关(r=0.4218,P<0.05),与心率变化无相关(r=0.0404,P>0.05)。运动引起运动神经传导速度增快,可能与运动本身及体温升高引起机体一些生理变化有关,它具有一定理论和应用价值。