宫颈癌及癌前病变中人乳头瘤病毒16载量与整合频率的研究

目的 研究人乳头瘤病毒(HPV)16病毒载量、基因组整合状态与癌前病变进展的关系,探讨HPV基因组整合频率作为宫颈病变程度标志物的可行性.方法 收集宫颈脱落细胞标本337例,通过PCR扩增、DNA测序分析筛选出HPV16单一感染的样品40例,利用实时定量PCR检测早期基因E2、E6和β-肌动蛋白的拷贝数,分析病毒载量和整合状态.结果 (1)校正后病毒载量在正常组、低度鳞状上皮内病变(LSIL)组、高度鳞状上皮内病变(HSIL)组和宫颈鳞癌组的中位数分别是0.11拷贝/细胞、18.55拷贝/细胞、44.63拷贝/细胞、7.6拷贝/细胞,正常组低于LSIL与HSIL组,宫颈鳞癌组低于HSIL组.(2)在正常、癌前病变(LSIL-HSIL)和宫颈鳞癌中E2/E6比值的中位数分别是0.93、0.84、0.64,E2/E6比值与病变的严重程度显著相关.(3)随着病变程度的升高,游离型比例逐渐降低:正常4/5、LSIL4/6、HSIL 10/16、宫颈鳞癌5/13,整合型(混合+整合)比例逐渐升高:正常1/5、LSIL 2/6、HSIL 6/16、官颈鳞癌8/13.2例完全整合的标本均为宫颈鳞癌.结论 HPV16病毒载量可能不是预测宫颈病变的理想指标.HPV16整合频率与宫颈病变严重程度正相关,可能可作为HPV16感染后宫颈上皮细胞病变发展的预估指标.

Abstract：

Objective To investigate the association between viral load, genomic integration frequency of HPV 16 and cervical carcinogenesis and assess the probability that HPV 16 integration frequency may be utilized as a marker for cervical cancer. Methods Forty cases of HPV16 single infection were selected from 337 cervical scrape samples by PCR (polymerase chain reaction) amplification and DNA sequencing. The copy numbers of E2, E6 and β-actin were quantified to evaluate the viral load and integration status by real-time PCR. Results ( 1 ) The median number of adjusted viral load of normal group, LSIL (low-grade squamous intraepithelial lesion) group, HSIL (high-grade squamous intraepithelial lesion) group and squamous cervical cancer group were 0. 11, 18. 55, 44. 63 and 7.6 copies per cell respectively. The viral load was higher in LSIL-HSIL group than that in normal group while lower in squamous cervical cancer group than that in HSIL group. (2) The median number of E2/E6 was 0. 93 in normal group, 0. 84 in precancerous group (LSIL-HSIL) and 0. 64 in squamous cervical cancer group respectively. It showed a descending trend with the progression of cervical disease. (3) With the disease development, the proportion of episomal form decreased ( normal group 4/5, LSIL group 4/6, HSIL group 10/16, cervical squamous cancer group 5/13 ) whereas that of integrated form ( mixed and totally integrated)increased (normal group 1/5, LSIL group 2/6, HSIL group 6/16, cervical squamous cancer group 8/13).Both totally integrated cases were cervical squamous cancer. Conclusion ( 1 ) HPV 16 viral load may not be an ideal marker to predict cervical carcinogenesis. (2) Due to a positive correlation between HPV16 genomic integration frequency and cervical neoplastic progression, HPV 16 integration frequency may be a potential marker of early diagnosis for cervical lesion progression.     

大蒜新素对宫颈病变组织HPV16E6基因表达的影响

[目的]研究大蒜新素对宫颈病变组织HPV16 E6基因表达的影响.[方法]运用PCR技术检测宫颈病变组织HPV16E6基因,表达阳性的病例120例随机分为安慰剂组、大蒜新素治疗组、阿昔洛韦治疗组,观察治疗前后宫颈病变情况、HPV转阴率.[结果]治疗后大蒜新素治疗组、阿昔洛韦治疗组宫颈局部情况转为正常者与安慰剂组相比有统计学意义(P<0.05);大蒜新素治疗组与阿昔洛韦治疗组的HPV转阴率比较差异无统计学意义(P>0.05),大蒜新素治疗组与安慰剂组,阿昔洛韦治疗组与安慰剂组的HPV转阴率比较差异均有统计学意义(P<0.05).[结论]大蒜新素治疗后宫颈病变情况减轻,局部组织结构恢复正常,机体的病毒清除率大有提高,值得在临床上推广.     

Hawthorn Extract Alleviates Atherosclerosis through Regulating Inflammation and Apoptosis Related Factors: An Experimental Study

Objective: To determine the effects of hawthorn extract on serum lipid levels, pathological changes in aortic atherosclerosis plaque, inflammatory factors, and apoptosis-related protein and mRNA expression in apolipoprotein E gene knockout(ApoE~(-/-)) mice. Methods: Thirty-six ApoE~(-/-) mice were fed with a high-fat diet starting at the age of 8 weeks. Mice were randomly divided into 3 groups by a random number table including model group, hawthorn extract group, and simvastatin group, 12 mice in each group. Twelve 8-week-old C57BL/6 mice were fed a basic diet and served as control. The mice in the control and model groups were administered 0.2 mL saline daily, the mice in the hawthorn extract and simvastatin groups were administered with 50 mg/kg hawthorn extract or 5 mg/kg simvastatin daily for 16 weeks. After 16 weeks, plasma lipids including total cholesterol(TC), triglyceride(TG), low-density lipoprotein cholesterol(LDL-C) and high-density lipoprotein cholesterol(HDL-C) were determined by an enzymatic assay. Aortic atherosclerotic lesions were observed by light microscopy, scanning and transmission electron microscopy, respectively. Plasma levels of monocyte chemoattractant protein-1(MCP-1), interleukin-1β(IL-1β), adiponectin(APN), and hypersensitive C-reactive protein(hs-CRP) were measured by enzyme-linked immunosorbent assay(ELISA). Protein and mRNA expressions of Bax and Bcl-2 in the aorta were assessed by Western blotting and quantitative real-time polymerase chain reaction(qR T-PCR), respectively. Results: Compared to the control group, the plasma levels of TC, TG and LDL-C were significantly increased and HDL-C were significantly decreased in the model group(P0.01). Compared to the model group, treatment with hawthorn extract significantly decreased the plasma levels of TC, TG, and LDL-C and increased the plasma level of HDL-C in ApoE~(-/-)mice(P0.01). The levels of MCP-1, IL-1β, and hs-CRP in the model group were significantly increased and APN was significantly decreased compared with the control group(P0.01). Compared to the model group, treatment with hawthorn extract decreased the levels of MCP-1, IL-1β, and hs-CRP and increased the APN level(P0.01). Compared to the control group, the protein and mR NA expression of Bax in the model group were significantly increased and the expression of Bcl-2 was significantly decreased(P0.01). Hawthorn extract also reduced the protein and mR NA expression of Bax and increased the Bcl-2 expression in the aorta(P0.01). Conclusion: Hawthorn extract has anti-atherosclerosis and stabilizing unstable plaque effects. The mechanism may be related to the inflammation and apoptosis signaling pathways.     

Prevalence of high-risk human papillomavirus infection in different cervical lesion among organized health-examination women in Shanghai, China

Background Cervical cancer is the second most common cause of death from cancer among women worldwide. Human papillomavirus （HPV） plays a central role in the etiology of cervical cancer. It is important to describe the prevalence of HPV infection in different types of cervical lesions and to explore the relation between HPV viral load and the severity of cervical lesions. Methods To describe the HPV infection prevalence and viral load in different age groups, we retrospectively investigated 6405 cases of women who were organized by their units to take health-examination. They were given Hybrid Capture II tests between January 2005 and December 2006. The correlation between HPV viral load and pathology was assessed. Results Overall HPV infection prevalence was 29.1% （1864/6405）, while in women 18-20 years old it was 54.4% （31/57）, the highest among all age groups. After declining rapidly, HPV prevalence stabilized at about 30.0% in women aged 30 and older. Of the 6405 women, 1483 women had a colposcopic biopsy and 33.2% （492/1483） were positive for HPV DNA. Twenty-one percent of women with a normal diagnosis （238/1095） had HPV infection, a statistically significantly lower prevalence than in women with cervical lesions, including those with cervical intraepithelial neoplasia （68.8% in CIN1,66.7% in CIN2, and 76.5% in CIN3） or with cervical cancer （94.1%）. The correlation coefficient between viral load and cervical lesion severity was 0.134, which was not statistically significant （P=0.075）. Viral load values in women with CINs and cervical cancer were calculated, and no significant differences were identified. Conclusions The prevalence of high-risk HPV infection among women attending hospitals for health-examination in Shanghai is similar to the worldwide rate. HPV viral load can distinguish cervical lesions from normal individuals but cannot adequately predict the severity of cervical lesions.     

Chinese Herbal Formula Ermiao Powder (二妙散) Regulates Cholinergic Anti-inflammatory Pathway in Rats with Rheumatoid Arthritis

Objective: To investigate the effect of Chinese herbal formula Ermiao Powder (二妙散, EMP) on the expression of cholinergic anti-inflammatory pathway in rats with rheumatoid arthritis (RA). Methods: Seventy two rats were randomly ivided into 6 groups according to body weight, including normal control group, collagen induced arthritis (CIA) group, three doses EMP groups, and methotrexate (MTX) group (n=12 per group). All of the rats except for those in the normal control group were given multipoint subcutaneous injection of bovine type Ⅱ collagen to establish a CIA model. Three EMP groups received a high- (4.5 g/kg), medium- (3.0 g/kg), and low- (1.5 g/kg) doses of EMP by intragavage, respectively. MTX group was injected intraperitoneally MTX at 0.9 mg/kg once a week as the positive control. The administration was 3 consecutive weeks. Joint swelling, arthritis index, and body weight changes in different experimental groups of rats were tested. The joint damage was evaluated by masson staining. Quantitative real-time polymerase chain reaction, Western blot, and immunohistochemistry (IHC) were performed to evaluate the expression of CHRNA7, encoding α 7 nicotinic acetylcholine receptor in the cholinergic anti-inflammatory pathway, in different tissues and their localization in the spleen and joints. Results: CHRNA7 expression levels were significantly higher in the joints and spleens of CIA group than those in normal control group (both P<0.05). Moreover, the CHRNA7 mRNA and protein levels in the spleen and joints of MTX and three doses of EMP groups were significantly lower than CIA group (all P<0.05). Compared with the MTX group, treatment with low-dose EMP resulted in significant reduction of CHRNA7 mRNA and protein expression levels (P<0.05 or P<0.01). IHC showed positive signals of CHRNA7 in the white pulp and red pulp of the spleens of rats; CHRNA7 was expressed on fibroblast-like synoviocytes, macrophages, and endothelial cells in the joints of rats, and the expression in the joints of low-dose EMP group was significantly lower than that in the CIA group (P<0.01). Conclusion: Cholinergic anti-inflammatory pathway was involved in the generation of the inflammatory reaction in CIA rats, and EMP exerted therapeutic effect on RA through cholinergic anti-inflammatory pathway.     

Effect of Wenhua Juanbi Recipe (温化蠲痹方) on Expression of Receptor Activator of Nuclear Factor Kappa B Ligand, Osteoprotegerin, and Tumor Necrosis Factor Receptor Superfamily Member 14 in Rats with Collagen-Induced Arthritis

Objective: To study the effect of Wenhua Juanbi Recipe(温化蠲痹方, WJR) on expression of receptor activator of nuclear factor kappa B ligand(RANKL), osteoprotegerin(OPG), and tumor necrosis factor receptor superfamily member 14(TNFRSF14, also known as LIGHT) in rats with collagen-induced arthritis(CIA). Methods: CIA rats were generated by subcutaneous injection of bovine collagen type-Ⅱ at the tail base. Sixty CIA rats were randomly assigned(10 animals/group) to: model, methotrexate(MTX)-treated(0.78 mg/kg body weight), and WJR-treated(22.9 g/kg) groups. Healthy normal rats(n=10) were used as the normal control. Treatments or saline were administered once daily by oral gavage. Rats were sacrificed at day 28 post-treatment and knee synovium and peripheral blood serum were collected. Toe swelling degree and expression of RANKL, OPG, and LIGHT were determined by Western blot and immunohistochemistry. Results: Compared with the normal group, toe swelling degree was significantly increased in the model group(P0.01). After treatment, toe swelling degree decreased significantly in the WJR and MTX groups compared with the model group(P0.01). Compared with the normal group, expression of RANKL and LIGHT were significantly increased and OPG significantly decreased in peripheral blood and synovium of the model group(P0.01). Conversely, RANKL and LIGHT expression were significantly reduced and OPG increased in the WJR and MTX groups compared with the model group(P0.01). No statistically significant difference existed between WJR and MTX groups. Conclusion: WJR likely acts by reducing RANKL expression and increasing OPG expression, thus inhibiting RANKL/RANK interaction and reducing LIGHT expression, thereby inhibiting osteoclast formation/activation to block bone erosion.     

Effect of All-trans Retinoic Acid on Airway Inflammation in Asthmatic Rats and Its Mechanism

The inhibitive effects of all-trans retinoic acid (ARTA) on airway inflammation in asthmatic rats and its mechanism on the basis of the regulation of nuclear factor kappaB (NF-kB) were explored. Thirty-two SD rats were randomly divided into 4 groups: control group, asthma group,dexamethasone treatment group and retinotic acid treatment group. The total and differential cell counts in the collected bronchoalveolar lavage fluid (BALF) were measured. The pathological changes in lung tissues were estimated by scoring. The expression of NF-kB inhibitor (IkBa), NF-kB,intercellular adhering molecule-1 (ICAM-1) in lung tissue was detected by immunohistochemical method. The results showed that in the two treatment groups, the total cell counts and proportion of inflammatory cells in BALF were significantly reduced, but there was no significant difference in differential cell counts in BALF between them. The pathological changes in lung tissues in the treatment groups were significantly attenuated as compared with asthma group. Except the epithelial injury in retinotic acid treatment group was milder than in dexamethasone treatment group, the remaining lesions showed no significant difference between them. In the two treatment groups, the expression of IkBa was increased, while the expression of NF-kB and ICAM-1 decreased with the difference between the two groups being not significant. It was concluded that the similar anti-inflammatory effects and mechanism of ATRA on airway in asthmatic rats to those of dexamethasone were contributed to the increase of cytoplasmic IkBa content and suppression of NF-kB activationand expression.     

SPECIFIC UP-REGULATION OF DNA POLYMERASE BY HUMAN PAPILLOMAVIRUS 16

Obje, etive To analyze how the infection of human papillomavirus 16 （ HPV16 ） affects expression of DNA poly- merase β（DNA polB） with the aim of probing the mechanism of over-expression of DNA polB in human cancers. Methods Four fragments of human DNA polB promoter were amplified and constructed into luciferase reporter vec- tor pGL-Basic, generating pGL-BP, pGL-BMH, pGL-BMS, and pGL-BAT constructs respectively, and co-transfected with HPV16 or HPV6 into Hep2 cells. Luciferase activity was assayed 48 hours after transfectiorL Semi-quantitative RT- PCR was used to measure mRNA expression of endogenous DNA polB. Immunohistochemistry and in situ hybridization were used to analyze DNA polB expression and HPV16 or HPV6 infection in 38 cases of cervical lesions respectively. Results With co-transfection of HPV16 and DNA polB promoter-driving reporters into Hep2 cells, pGL-BP re- porter in full-length DNA polB promoter presented markedly elevated luciferase activities （ P 〈 0.05 ）. However, the other three mutant reporters： pGL-BMH, pGL-BMS, and pGL-BAT, generated no reporting activities in the presence of HPV16 （P 〉0.05 ）. On the contrary, all of polB promoter reporters were little stimulated in co-transfection of HPV6 （P 〉0.05 ）. The transfection of HPV16 could enhance the endogenous polB mRNA expression compared with that of HPV6 （3.42 vs. 0.80, P 〈0.05）. The DNA polB expression was found in 8 of 10 HPV16-positive cervical intraepi- thelial neoplasia grade Ⅲ（ CIN Ⅲ） cases, while was only found in 3 of 11 HPV6-positive condyloma accuminatum cases, but was negative in all chronic cervicitis cases. The correlation of DNA polB expression with HPV16 infection in cervical lesions was significant （ P 〈0.05 ）. Conclusions HPV16 is able to specifically stimulate the expression of DNA polB in human epithelial cells through interaction with the core upstream regulatory sequences of DNA polB promoter. Over-expression of DNA polB might be an explanation for the molecular mechani     

麻疹的双相移位研究进展

近年新生儿、婴儿、成人麻疹患者逐年增加,临床表现一般仍较典型,成年人麻疹患者全身中毒症状较重。麻疹抗体检测结果阳性是主要的诊断依据。麻疹发病的双相移位的机理可能是,免疫保护力不足,婴儿出生时麻疹抗体力低。孕期母传胎的麻疹抗体减弱,母经乳汁传给婴儿的抗体减弱,成人麻疹抗体水平逐年下降。预防措施是怀孕前给予育龄妇女麻疹疫苗接种,鼓励母乳喂养,麻疹疫苗计划免疫适当提前,在成人追加麻疹疫苗的免疫,加强病毒变异的研究等。     

正常增殖期子宫内膜细胞增生状态的定量观察

以＾３氢－胸腺嘧啶核苷放射自显影法及ＨＥ染色，观察并分别测定了１８例正常子宫内膜增殖中期，１５例增殖晚期的腺上皮细胞或间质细胞的标记指数、分裂指数。结果显示：子宫内膜增殖晚期腺上皮细胞或间质细胞之ＬＩ均明显高于增殖中期。同时，增殖晚间质细胞之ＭＩ也明显高于增殖中期，即此两种细胞在增殖晚期中增生明显，其增生状态初步获得了定位定量测定的正常值。     

人工全髋关节置换术的手术配合

目的：介绍人工全髋关节置换术的手术配合做法;方法：主要在手术配合的六个方面,解决防感染、防栓塞等问题。结果：30例人工全髋关节置换术均获成功,结论：手术配合是护士责任心和基本功的全面体现,对提高手术效果有至关重要的影响。     

尿微量白蛋白检测在继发性肾脏疾病中的临床意义

目的探讨尿微量白蛋白（m-Alb）检测在继发性肾脏疾病中的临床意义。方法采用Beckman Immage全自动特种蛋白分析仪对糖尿病组、高血压组、心脏病组患者进行了m-Alb测定,同时与健康组结果作对比。结果m-Alb检测糖尿病组为3.7±5.26mg/dl,高血压组为7.5±8.18mg/dl,心脏病组为7.8±3.76mg/dl,健康组为0.66±0.48mg/dl,各试验组m-Alb增高百分率为糖尿病组48.9%,高血压组37.5%,心脏病组26.9%。结论尿蛋白阴性的糖尿病、高血压、心脏病患者进行m-Alb检测,可以监测病程的进展。     

尿液pH值对红细胞检验影响的探讨

[目的 ]通过尿液 pH值对红细胞检验影响的观察 ,更加科学、准确地诊断血尿和血红蛋白尿。[方法 ]采用干化学分析仪检测和尿液显微镜红细胞计数 ,观察 180例正常人尿标本加入正常人血标本后 ,不同 pH值 ,不同时间内 ,观察红细胞溶解情况。 [结果 ]pH <5 .5以下时 ,随着时间的延长 ,红细胞溶解现象明显。 1h后观察有显著性差异 (P <0 .0 5 ) ;2h后有非常显著性差异 (P <0 .0 1)。[结论 ]pH <5 .5时对红细胞计数影响较大 ,易致红细胞发生溶解现象 ,出现假性血红蛋白尿 ,对血尿和血红蛋白尿很难区分 ,给临床诊断造成不便 ,更易引起漏诊和误诊。     

80例局限性银屑病误诊原因分析

本文报告80例局限于小腿或手或足的银屑病。均经皮肤组织病理检查确诊。因部位比较特殊。受多种理化因素影响,使皮疹形态发生轻重程度不同的变化,常看不到典型损害,因而误诊为神经性皮炎,湿疹,慢性皮炎及癣等。作者对误诊原因进行了分析后,提出了鉴别诊断方法。     

腰椎间盘突出症再次手术的原因分析

目的解决腰椎间盘突出症手术中神经压迫。方法对1980～1998年再手术资料进行统计分析,讨论分析再手术原因,再次手术前影像学检查,观察病理变化以确定再手术方法。结果对11例随访6个月～1年,优7例(68．4％),良3例(36．8％),差1例(2．8％)。结论初次手术前详细查体和分析X线片,术中用导尿管和神经剥离探查,尽量避免髓核遗留,手术范围不宜太大,尽量减少对软组织和脊柱结构的破坏,避免形成硬膜囊与神经根粘连而致单纯形疤痕。