胃癌患者胃粘膜无芽胞厌氧菌分析及药物敏感试验

对98例具有消化不良症状的患者做胃镜检查,取胃粘膜组织进行细菌分离培养,结果表明,不同的消化道疾病,胃粘膜微生物群均发生不同的改变,其中胃癌患者改变最明显,胃内微生物检出率最高,无芽胞厌氧菌检出率87.88％（29／33）,其中产亚硝酸优杆菌检出率为57.60％（19／33）,硝酸盐还原试验阳性菌株占72.70％（24／33）。这些细菌和胃癌的关系有待深入研究。对19株产亚硝酸优杆菌做药物敏感试验,结果显示,该菌对青霉素类药物均耐药,对氯霉素,红霉素,庆大霉素敏感。     

消化道疾病患者胃微生态环境的改变

本文对５９例具有消化不良症状的患者做了内镜检查。取胃粘膜组织进行细菌的分离培养,镜检,测定胃液中的ｓＩｇＡ的含量及ｐＨ。结果表明,不同消化道疾病,胃的微生态环境会发生不同的改变。其中胃癌患者改变最明显,胃内微生物检出率高,ｐＨ均大于４,ｓＩｇＡ含量１４０．５７±３７．４５μｇ／ｍｌ,幽门螺杆菌（ＨＰ）的检出率为６６．７％;溃疡组,ＨＰ的检出率为７９．２％,其它微生物检出率较胃癌组低,ｓＩｇＡ含量１３３．８０±６５．８４μｇ／ｍｌ;胃炎组,微生物的检出率较低,ｓＩｇＡ含量７１．００±６０．３８μｇ／ｍｌ,ｐＨ大部分在２一４。     

性病后慢性前列腺炎病原微生物分析

本文对性病后慢性前列腺炎病原微生物进行了研究。９０例患者前列腺液支原体检出率为２４．４４％（２２／９０）,其中解脲支原体为２２．００％（１８／９０）,人型支原体为４．４４％（４／９０）。另一组２３２例患者进行前列腺液细菌培养鉴定,总检出率为４２．７％（９９／２３２）,以金黄色葡萄球菌为主２４．５％（５７／２３２）,其它菌依次为表皮葡萄球菌７．３％（１７／２３２）,肠球菌４．３％（１０／２３２）,非发酵菌２．６％（６／２３２）,肠杆菌科细菌２．２％（５／２３２）和Ａ群链球菌１．７％（４／２３２）。作者认为,性病后慢性前列腺炎可能为急性尿道炎期,由于治疗不彻底或忽略非特异性性病病原菌的治疗而使条件致病菌上行感染所致。     

武汉地区不同人群真菌带菌调查

本文报道了湖北省武汉地区不同人群带真菌情况的调查结果。共检查２０５１人,调查组带菌率为８６．４６％;对照组（消毒后取样检验的住院病人）为３４．６５％,两者差异显著。不同部位检出率不同,调查组在腔孔部位阳性检出率最高,为７８．２１％,对照组以粘膜部位较高,为２２．７７％,并带念珠菌属的真菌为主,为８８．３７％。     

眼镜蛇神经毒素的急性毒性实验研究

目的　研究眼镜蛇神经毒素 （ Ｃｏｂｒａ ｎｅｕｒｏｔｏｘｉｎ, Ｎ Ｔ） 的急性毒性和蓄积毒性。方法　测 Ｎ Ｔ 对小鼠的 Ｌ Ｄ５０ ; 对大鼠、狗的１ 次性最小中毒剂量和最大安全剂量; 计算 Ｎ Ｔ 在小鼠、狗体内的２４ｈ 蓄积率。结果　 Ｎ Ｔ 经静注、肌注、腹腔、皮下 ４ 种途径给药对小鼠的 Ｌ Ｄ５０ 分别是 （１９５±９５） μｇ／ｋｇ、（１５６±８５） μｇ／ｋｇ、（１５１±１９） μｇ／ｋｇ、（１８４±８５） μｇ／ｋｇ, 对小鼠的最小致死剂量为９７５μｇ／ｋｇ。 Ｎ Ｔ 对大鼠、狗的１ 次性中毒剂量分别为５４μｇ／ｋｇ 和３４μｇ／ｋｇ。对小鼠、大鼠和狗的安全剂量分别为８１５μｇ／ｋｇ、４２μｇ／ｋｇ和３０μｇ／ｋｇ, 分别约为人临床用剂量 （７０μｇ／５０ｋｇ·ｄ－ １ ） 的５８２、３０ 和２１ 倍。 Ｎ Ｔ 在小鼠、狗体内的２４ｈ蓄积率分别为 ５７％ 和３０％ 以上。结论　 Ｎ Ｔ 在使动物中毒的剂量下有广泛的安全范围; Ｎ Ｔ 在动物体内存在弱蓄积毒性。     

幽门螺杆菌L型实验动物感染的研究

本文用２～５×１０９个幽门螺杆菌临床分离株细菌型与Ｌ型口饲感染了不同种的小鼠（ＫＭ种,Ｃ５７ＢＬ／６Ｎ系）,于感染后９０天处死小鼠。组织学检查表明,口服细菌型与Ｌ型小鼠多数出现胃肠粘膜组织的慢性间质性炎症（１５／１８,２４／３４）,表现为淋巴细胞和单核巨噬细胞的浸润,同时引起肝脏细胞的轻微毒性变化。免疫组化技术（ＡＢＣ法）证实,这些病变确由幽门螺杆菌感染引起（ＨＰ抗原检出率１２／１５和１９／２４）。在形态上,两组细菌均主要以圆球体形式在胃粘膜存在和潜伏。提示ＨＰ－Ｌ型仍保留一定的毒力因素,是慢性胃炎、十二指肠溃疡迁延不愈和反复发作的潜在因素,与胃癌的发生发展也有一定的关系。     

72例肝脓肿厌氧菌培养结果分析

报告７２例肝脓肿患者脓液厌氧和需氧培养,结果４４例培养阳性、总检出率６１．１％,其中单纯厌氧菌和单纯需氧菌检出率,分别占阳性标本的２２．７％和３４．１％,混合阳性占４３．２％。共检出细菌７２株（厌氧菌３４株,需氧菌３８株）,其中以消化链球菌（５０．０％）和大肠埃希氏菌（３４．２％）多见。并对诱发原因,细菌种类和药敏试验进行了讨论。     

血标本检出420株菌的菌谱分布及药敏分析

本文报告中山医科大学附一院从１９９５年１０月至１９９７年８月应用Ｂａｃｔ／Ａｌｅｒｔ血培养仪检测３６００份血液培养结果,阳性标本４１４份,其中６份为复数菌,菌株数４２０,其中革兰阳性球菌（Ｇ＋Ｃ）２３６株,革兰阴性杆菌（Ｇ－Ｂ）发酵菌７２株,Ｇ－Ｂ非发酵菌３４株,真菌５３株,革兰阳性杆菌（Ｇ＋Ｂ）１３株,革兰阴性球菌（Ｇ－Ｃ）２株,厌氧菌１０株。主要的细菌分离率由高到低为凝固酶阴性葡萄球菌（ＳＣＯＮ）１７８６％,金黄色葡萄球菌（ＳＡ）１２８６％,α—溶血性链球菌９５２％,肠球菌属９０５％,大肠埃希氏菌８１％,微球菌属６４３％,铜绿假单胞菌４２９％和沙门氏菌３１％。药敏结果显示Ｇ＋Ｃ中万古霉素的耐药率最低,Ｇ－Ｂ却对各种药物表现出不同的耐药率,其中对泰能和头胞噻甲肟的耐药率较低。     

应用大规模测序技术和生物信息学研究造血干/祖细胞的基因表达及新基因的识别和克隆

从新生儿脐血和成人骨髓中分选出造血干／祖细胞（ＨＳＣ／ＨＰＣ）,构建成ｃＤＮＡ文库,对其进行大规模表达序列标签（ＥＳＴ）测序,通过生物信息学等手段分析基因表达谱,并进行新基因的全长ｃＤＮＡ克隆。在所测的１０５１２条可分析ＥＳＴ序列中,有９８６６条来自脐血ＣＤ３４＋细胞,其中４６９７条（４７６％）为已知基因,２６０３条（２６４％）为已知ＥＳＴ,１４１５条（１４３％）代表未知ＥＳＴ。在已知基因中,８２％基因与造血相关,２２７％涉及细胞代谢、结构和迁移,１３０％与细胞分裂和防御相关,２６２％与ＲＮＡ、蛋白质的合成相关,１０６％和细胞信号传递有关。对一些已知和未知的ＥＳＴ,综合测序、生物信息学等方法,进行全长克隆,已获得２３个新基因的全长ｃＤＮＡ。     

HIV-1型核衣壳蛋白P24截短体在大肠杆菌中的表达,纯化和鉴定

用聚合酶链式反应（ＰＣＲ）从ＨＩＶ １ｇａｇ基因中扩增出衣壳蛋白Ｐ２４截短体（ｔＰ２４）基因,插入质粒ｐＧＥＸ ４Ｔ３中构成重组表达质粒ｐＧＥＸ ｔｐ２４,将ｐＧＥＸ ｔｐ２４转化大肠杆菌ＢＬ２１后获得了高效表达,表达量占菌体总蛋白量的３４３８％。经Ｇｌｕｔａｔｈｉｏｎｅ－Ｓｅｐｈａｒｏｓｅ４Ｂ亲和层析纯化的截短体Ｐ２４纯度为９２７７％。纯化的截短体Ｐ２４在间接ＥＬＩＳＡ和免疫印迹检测ＨＩＶ抗体阳性血清和正常人血清中,具有很高的抗原特异性和免疫反应性。     

胃癌患者肠道菌群的分布特点分析

目的应用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)基因指纹图谱技术,检测胃癌患者肠道菌群的变化情况,探讨肠道菌群与胃癌发生发展的关系。方法选取7例符合临床诊断标准的胃癌患者和相同年龄段的3例健康者的粪便标本,提取粪便总DNA。通过PCR-DGGE技术获得菌群指纹图谱,进行相似性、多样性分析。结果胃癌患者肠道菌群与健康对照组相比,其肠道菌群构成发生显著性变化。其中,条件致病菌唾液乳杆菌、唾液链球菌、大肠埃希菌数量均增加。益生菌直肠真杆菌、Dorea longicatena、柔嫩梭菌群数量均较少。结论胃癌患者肠道菌群的多样性和构成与健康对照相比发生了显著变化,该特点对于胃癌的预测提供了实验依据。     

31例胃癌患者胃液SIgA及胃粘膜产亚硝酸真杆菌测定结果分析

探讨了胃癌病变局部免疫功能及胃粘膜细菌分布与发病机制的关系.采用放射免疫法对31例胃癌患者(实验组)与56例其它胃病患者(实验组)进行胃液SIgA含量的测定并对胃粘膜病变部位进行细菌分离培养、涂片染色镜检.结果显示,实验组较对照组胃液SIgA含量显著增多(p<0.05),胃粘膜产亚硝酸真杆菌阳性检出率亦明显升高(p<0.01).胃癌患者胃粘膜局部免疫功能失常,其发病可能与产亚硝酸真杆菌有关,胃液SIgA的测定对估价预后有一定意义.     

Chronic gastritis associated with Helicobacter pylori. Correlation between histological and bacteriological findings.

Biopsy specimens of gastric and duodenal mucosa from 326 patients were examined bacteriologically and histologically to determine the correlation between chronic gastritis and H. pylori colonization. H. pylori was identified in 111 (66.5%) patients with evidence of chronic gastritis and in 97 (82.2%) individuals who had gastritis associated with other pathology (gastric o duodenal ulcer, carcinoma o bulboduodenitis). The spiral bacteria was found more frequently in specimens with chronic superficial gastritis (88/107) and no significant difference was observed between the grade of activity of gastritis and H. pylori colonization. Giemsa stain was the most suitable method for detecting H. pylori in histological sections. By electron microscopy the microorganism was seen on the surface of the gastric mucosa, beneath the mucous layer, and more occasionally in intercellular junctions and the gastric pit.     

Characterization of bacterial community in the stomach of yellow catfish (<Emphasis Type="Italic">Pelteobagrus fulvidraco</Emphasis>)

In this study, the traditional culture-based technique and the 16S rDNA sequencing method were used to investigate the characterization of bacterial community in the stomach contents and mucus of yellow catfish (Pelteobagrus fulvidraco). The culture-based technique disclosed that the average bacterial numbers in the gastric contents and mucus were 5.79 × 10⁷ cfu/g (cfu: colony forming unit) and 1.89 × 10⁵ cfu/g, respectively. Several different bacteria were obtained from gastric contents, including species from genera Bradyrhizobium, Phyllobacterium, Plesiomonas, Hafnia, Edwardsiella, Pseudomonas, and Bacillus. However, only two species were isolated from the gastric mucus, including species from genera Plesiomonas and Aeromonas. Forty-five phylotypes were observed from the 65 positive clones from the stomach contents (library SC); nineteen phylotypes were detected from the 45 clones from the stomach mucus (library SM). Further analyses revealed that the fish stomach harbored characteristic microbiota, where Firmicutes was dominant, followed by Proteobacteria and Bacteroidetes and Fusobacteria. This characterization of bacterial community is markedly different from that of the fish intestine, where Proteobacteria is predominant, followed by Fusobacteria and Firmicutes. Chloroflexi (1.5%) was only found in the library SC, while Actinobacteria (4.4%) was only found in the library SM, suggesting that microbiota of GI contents was quite different from that of GI mucus. In addition, several species of bacteria found in the stomach may be potentially opportunistic pathogens, indicating that fish digestive tract is a reservoir for many nosocomial pathogens.     

Low prevalence of Helicobacter pylori-negative gastric cancer among Japanese

Background and Aims: The true prevalence of Helicobacter pylori‐negative gastric cancer (HpNGC) is unknown. We attempt to clarify the prevalence and clinicopathologic features of HpNGC in Japanese. Methods: Helicobacter pylori infection was detected by antibody titer and microscopic observation. In addition, we confirmed the lack of endoscopic atrophy and histologic gastritis. In these cases, we added urea breath test or rapid urease test to confirm the absence of H. pylori. The mucus phenotype of gastric cancer tissue was also evaluated by immunohistochemistry. Results: We screened 3161 gastric cancer cases from 1996 to 2010, and 21 cases were regarded as H. pylori negative. Clinically, patients with HpNGC were younger than patients with H. pylori‐positive gastric cancer (controls), and revealed a lack of male dominancy. Histologically, diffuse type was frequently found. All patients examined were pepsinogen negative. Among HpNGC cases with endoscopic resection, the depressed macroscopic appearance was dominant. The prevalence of HpNGC was calculated as 0.66% (95% confidence interval = 0.41–1.01). The mucus phenotype of HpNGC was similar to that of the controls. Conclusion: The prevalence of HpNGC is very low and its pathological characteristics are different from common gastric cancer.     

血清蛋白质指纹图谱诊断早期胃癌临床意义

目的：应用SELDI蛋白质芯片检测胃癌患者血清蛋白质指纹图谱,筛选候选肿瘤标志物以建立诊断模型。方法：表面加强激光解吸电离-飞行时间质谱（SELDI-TOF-MS）技术及其配套蛋白质芯片检测34例胃癌患者（Ⅰ／Ⅱ期12例与Ⅲ／Ⅳ期22例）和30例健康人的血清蛋白质组图谱,运用判别分析处理数据筛选标志物并建立诊断模型。结果：2046m／z、1179m／z、1817m／z、1752m／z和1588m／z等5个蛋白质峰组合所构建的诊断模型能达到鉴别胃癌患者和健康人的最佳诊断效果,特异度94．1％（32／34）,灵敏度93．3％（28／30）。单个4665m／z蛋白质峰诊断模型可达到鉴别Ⅰ／Ⅱ期与Ⅲ／Ⅳ期胃癌效果,其特异度91．6％（11／12）,灵敏度95．4％（21／22）。结论：该方法在胃癌的诊断尤其是早期诊断方面具有一定价值,值得进一步研究。     

Physiological recycling of endogenous nitrate by oral bacteria regulates gastric mucus thickness

BackgroundInorganic nitrate from exogenous and endogenous sources is accumulated in saliva, reduced to nitrite by oral bacteria and further converted to nitric oxide (NO) and other bioactive nitrogen oxides in the acidic gastric lumen. To further explore the role of oral microbiota in this process we examined the gastric mucus layer in germ free (GF) and conventional mice given different doses of nitrate and nitrite.MethodsMice were given either nitrate (100 mg/kg/d) or nitrite (0.55–11 mg/kg/d) in the drinking water for 7 days, with the lowest nitrite dose resembling the levels provided by swallowing of fasting saliva. The gastric mucus layer was measured in vivo.ResultsGF animals were almost devoid of the firmly adherent mucus layer compared to conventional mice. Dietary nitrate increased the mucus thickness in conventional animals but had no effect in GF mice. In contrast, nitrite at all doses, restored the mucus thickness in GF mice to the same levels as in conventional animals. The nitrite-mediated increase in gastric mucus thickness was not inhibited by the soluble guanylyl cyclase inhibitor ODQ. Mice treated with antibiotics had significantly thinner mucus than controls. Additional studies on mucin gene expression demonstrated down regulation of Muc5ac and Muc6 in germ free mice after nitrite treatment.ConclusionOral bacteria remotely modulate gastric mucus generation via bioactivation of salivary nitrate. In the absence of a dietary nitrate intake, salivary nitrate originates mainly from NO synthase. Thus, oxidized NO from the endothelium and elsewhere is recycled to regulate gastric mucus homeostasis.     

Cytodiagnosis of Campylobacter pylori in Papanicolaou-stained imprints of gastric biopsy specimens.

The use of Papanicolaou-stained touch preparations of gastric antral biopsies for the identification of Campylobacter pylori was examined using specimens obtained from 63 consecutive patients with endoscopic evidence of antral gastritis, with the results compared to routine histologic examination and Warthin-Starry silver staining. Organisms were readily identifiable in the Papanicolaou-stained imprints of the gastric mucus. The sensitivity in detecting organisms was 92.5% for the Warthin-Starry-stained sections, 71.4% for the Papanicolaou-stained imprints and 100% for both techniques combined. False-negative imprints were attributed to poor smears and/or the submission of duodenal tissue rather than antral biopsies. Properly performed touch preparations stained by the Papanicolaou method are a cost-effective adjunct to Warthin-Starry-stained section for improving the sensitivity of gastric biopsies for the diagnosis of C pylori.     

Quantification of the Iodine Content of Perigastric Adipose Tissue by Dual-Energy CT: A Novel Method for Preoperative Diagnosis of T4-Stage Gastric Cancer

This study investigated the utility of quantifying iodine concentration (IC) in perigastric adipose tissue, using dual-energy computed tomography (DECT), for the detection of T4a-stage gastric cancer. Fifty-four patients with gastric cancer were enrolled at the Fourth Hospital of Hebei Medical University between January and June 2013. Patients were imaged preoperatively with conventional computed tomography (CT) scans and DECT, and the IC in perigastric fat adjacent to the tumor calculated from arterial phase (AP) and portal venous phase (PVP) images. The patients subsequently received surgical treatment (gastrectomy), and histologic analysis of resected specimens was used as a ‘gold standard’ reference for cancer staging. Receiver operating characteristic (ROC) curve analysis was employed to assess the utility of DECT for identifying T4a-stage gastric cancer, with optimal IC thresholds determined from the area under the ROC curve (AUC). Postoperative histology revealed that 32 patients had serosal invasion (group A), and 22 did not (group B). The accuracy of conventional CT for distinguishing stage T4 from non-T4 stages was 68.5% (37/54). IC was significantly higher in group A than in group B (AP: 0.60±0.34 vs. 0.09±0.19 mg/mL, p<0.001; PVP: 0.83±0.41 vs. 0.27±0.21 mg/mL, p<0.001). The sensitivity, specificity and AUC for detecting serosal invasion were 77.1%, 79.2% and 0.89 at an IC threshold of 0.25 mg/mL for AP images; and 80.0%, 79.2% and 0.90 at an IC threshold of 0.45 mg/mL for PVP images. These results indicated that Iodine quantification in perigastric fat using DECT is an accurate method for detecting serosal invasion by gastric cancer.