目的 探讨血清鳞状细胞癌相关抗原(SCC-Ag)联合白细胞介素-6(IL-6)、转化生长因子-β(TGF-β)检测对宫颈癌的价值.方法 选取2011年12月-2016年12月收治的58例宫颈癌患者为研究组,另选取2015年12月-2016年12月同院收治的子宫良性疾病且经术后病理检查证实宫颈正常的41例患者为对照组.两组均于清晨空腹采集外周静脉血3ml,分离血清,采用酶联免疫吸附法测定血清TGF-β、SCC-Ag及IL-6水平.比较两组患者血清TGF-β、SCC-Ag、IL-6水平,研究组不同临床分期及不同分化程度患者血清SCC-Ag、IL-6、TGF-β水平,分析血清SCC-Ag与IL-6、SCC-Ag与TGF-β的相关性.结果 研究组患者血清IL-6、TGF-β、SCC-Ag水平均显著高于对照组,差异均有统计学意义(均P＜0.05).研究组Ⅰ～Ⅱ期患者血清SCC-Ag、IL-6、TGF-β水平低于Ⅲ～Ⅳ期患者,差异均有统计学意义(均P＜0.05).研究组高分化患者血清SCC-Ag、IL-6、TGF-β水平低于中分化和低分化患者,中分化患者血清SCC-Ag、IL-6、TGF-β水平低于低分化患者,差异均有统计学意义(均P＜0.05).研究组患者血清SCC-Ag与IL-6、SCC-Ag和TGF-β均呈正相关(r=0.371,P＜0.05;r=0.497,P＜0.05).结论 宫颈癌患者的血清SCC-Ag、IL-6、TGF-β水平显著高于正常人,且随着病情加重水平上升显著,血清SCC-Ag与IL-6、SCC-Ag与TGF-β均呈正相关.

目的 研究脓毒症患者血清来源的外泌体对巨噬细胞极化的影响.方法 人单核细胞系的U 937细胞经佛波酯(PM A)刺激12 h后,分别用脂多糖(L PS)、白细胞介素(IL)-4、健康者血清来源外泌体和脓毒症患者血清来源外泌体刺激48 h,然后收集U937细胞并离心取上清液,用实时荧光定量PCR(qPCR)检测趋化因子(c-x-c模体)配体(CXCL)10、肿瘤坏死因子-α(TNF-α)、白细胞分化抗原(CD)206、肿瘤生长因子-β(TGF-β)和IL-1β的表达,用ELISA法检测培养上清液中CXCL10、TNF-α、IL-1β、和TGF-β的水平.结果 PMA诱导U 937细胞向巨噬细胞样细胞转化.这些巨噬细胞样细胞经L PS刺激后有明显较长的突起生成,而且XCL10、TNF-α、IL-1β表达水平增高,而IL-4刺激后细胞形态无明显变化,CD206和TGF-β表达水平增加.脓毒症患者血清来源外泌体刺激后细胞有明显较长的突起生成,而且CXCL10、TNF-α、IL-1β表达水平增高;健康者血清来源外泌体未明显影响细胞形态和CXCL10、TNF-α、IL-1β的表达.结论 脓毒症患者血清来源外泌体促进巨噬细胞向M1型极化.

Objective: To investigate the effects of polygonum capitatum on IL-17 and IL-23 levels in rats with Helicobacter pylori( H. pylori)-induced gastritis. Methods: Rats were devided into three groups including blank group,model group and polygounum capitatum treating group. The rats in the latter two groups were treated with H. pylori standard bacteria to establish the model of H. pylori chronic gastritis. After the modeling,the medication group was given polygonum capitatum with the dose of1. 72 g extract/( kg·d) by gavage for 2 weeks. Then normally feeding 2 weeks later,after that,rats were sacrificed,the gastric mucosa and abdominal aortic blood were collected,pathological changes were examined by HE staining,the levels of serum IL-17 and IL-23 were measured by enzyme-linked immunosorbent assay. The relative transcript levels of IL-17 and IL-23 mRNA in gastric mucosa were determined by real time PCR. Rusult: Compared with model group,the gastritis in treatment group alleviated remarkably and the pathological scores decreased statistically. The level of serum IL-17 and relative expression of mucosa IL-17 mRNA reduced obviously after polygonum capitatum therapy as well. The level of serum IL-23 also significantly decreased in polygonum capitatum group. Conclusion: Polygonum capitatum could reduce H. pylori-induced gastritic mucosal injury,which may be related to inhibiting IL-17 and IL-23 secretion.

Objective To determine the effect of epithelial-mesenchymal transition (EMT) of renal tubular epithelial cells induced by uric acid,and investigate the underlying mechanism.Methods Human renal tubular epithelial HK-2 cells were cultured in vitro and incubated with different concentrations of uric acid for 48 h.The effects of uric acid on the morphology of HK-2 cells were observed and recorded with the aid of inverted microscope.The protein levels of fibronectin,α-smooth muscle actin (o-SMA) and E-cadherin were detected by Western blotting,while the mRNA expression levels of IL-1β,IL-6 and TNF-α were detected by real-time PCR.Immunofluorescence assay was used to detect the changes of epithelial cell marker cytokeratin and mesenchymal cell marker vimentin.Results Compared with untreated HK-2 cells,treatment of 15 mg/dL uric acid for 48 h induced the HK-2 cells turned into long-spindled shape,with markedly increased intercellular space.The protein levels of mesenchymal marker fibronectin and o-SMA were significantly increased,while that of the epithelial cell marker E-cadherin was significantly decreased (P ＜0.05).High dose of uric acid enhanced the expression levels of IL-1β and TNF-α (P ＜ 0.05) and induced activation of TLR4/NF-κB signaling pathway in HK-2 cells (P ＜ 0.05).Targeted inhibition of NF-κB signaling pathway significantly suppressed EMT induced by uric acid in HK-2 cells.Conclusion High uric acid induces EMT in renal tubular epithelial cells by activation of the TLR4/NF-κB signaling pathway,and the targeted intervention of NF-κB signaling pathway can significantly suppress uric acid induced renal interstitial fibrosis.

目的 了解贵州省育龄妇女弓形虫感染状况并鉴定弓形虫基因型,为贵州省育龄妇女弓形虫感染的防治提供依据.方法 采用酶联免疫吸附试验检测729例育龄妇女外周血标本弓形虫循环抗原(CAg)、抗弓形虫抗体IgG、IgM,利用聚合酶链反应扩增弓形虫特异性基因片段(529 bp高重复序列)进行核酸检测,阳性标本则用多重巢式—聚合酶链式反应-限制性片段长度多态性分析方法对弓形虫进行基因分型.结果 479例孕妇外周血血浆中CAg、IgG和IgM阳性率分别为0.63％、7.10％和0.84％,总阳性率为8.56％.其中正常妊娠组样本弓形虫CAg、IgG和IgM阳性率分别为0.56％、4.47％和0.56％,有异常妊娠史组样本弓形虫CAg、IgG和IgM阳性率分别为0.83％、14.87％和1.65％,健康对照组中样本IgG阳性率为4.00％,未发现弓形虫CAg及IgM抗体.正常妊娠组1例、异常妊娠史组2例标本弓形虫核酸检测呈阳性,弓形虫基因分型为非典型虫株,即Chinese 1型(Toxo DB#9).结论 有异常妊娠史的孕妇弓形虫感染率明显高于正常妊娠孕妇及健康人群,提示异常妊娠与弓形虫感染密切相关,Chinese 1型弓形虫虫株可能是贵州育龄妇女人群中感染流行的优势虫株.

Objective:To survey on Toxoplasma gondii infection in depressed patients in Guizhou Province and identify the genotype of T. gondii. Methods:Enzyme-linked immunosorbent assay (ELISA) was applied to detect the T. gondii-specific antibodies IgG, IgM and circulating antigens (CAg) of T. gondii in 141 patients and 150 healthy subjects. The specific repeated DNA fragment (529 bp) of T. gondii was amplified by PCR. The genotype of T. gondii was determined by multiplex multilocus nested polymerase chain reaction-restriction fragment length polymorphism (Mn-PCR-RFLP). Results:ELISA showed that the positive rate of anti-T. gondii antibody in depressed patients and healthy subjects was 21.3%（30/141） and 7.3%（11/150）, respectively. The positive rate of IgG in depressed patients was 18.4% （26/141）, significantly higher than that in healthy subjects （7.3%, 11/150）（P<0.05）. The positive rate of IgM and CAg in depressed patients was both 1.4% （2/141）, while these were not found in healthy subjects. PCR revealed one patient positive for T. gondii, whose genotype was further identified to be Toxo DB #9（Chinese 1 type） by Mn-PCR-RFLP. Conclusion:The positive rate of T. gondii is higher in depressed patients than in the healthy subjects in Guizhou Province. The genotype of T. gondii detected in one depressed patient is the Chinese 1 type.

Objective:To investigate the effect of Polygonum capitatum on gastric mucosal gastrin(Gas) and somatostatin(SS) expression of SD rats infected by H.pylori and to explore the mechanism of Polygonum capitatum against H.pylori infection gastritis.Methods:42 rats were divided into model group of 26 rats and blank control group of 16 rats.Establishment of H.pylori rat model of chronic gastritis was established in rats by intragastric administration of H.pylori standard strain.After successful modeling,rats were divided into model group and Polygonum capitatum group.Polygonum capitatum group was given 1.72 g extractum/kg dose of Polygonum capitatum by intragastric administration while model group and blank group were given equal volume of saline by intragastric administration.After 2weeks of treatment,the rats were killed,the gastric mucosa was taken and the histological changes of gastric mucosa were observed by HE staining.Immunohistochemical staining was used to observe the changes of Gas positive cells and SS positive cells in gastric mucosa.The expression levels of mRNA Gas and mRNA SS in gastric mucosa were detected by in situ hybridization.Results:Compared with the model group,Polygonum capitatum can significantly alleviate the inflammation of the gastric mucosa of rats infected with H.pylori,and the pathological score decreased significantly(P<0.05).The number of Gas positive cells of gastric mucosa in Polygonum capitatum group significantly decreased and the expression level of Gas mRNA decreased significantly.The difference was statistically significant(P<0.05).There was no statistically significant difference in the number of SS positive cells and mRNA SS expression level between the model group and Polygonum capitatum group(P>0.05).Conclusion:Polygonum capitatum can repair gastric mucosal damage in chronic gastritis infected with H.pylori,and its mechanism may be related to improving the stomach intestine hormone regulation.

Background Toxoplasma gondii is an obligate, intracellular protozoan that infects almost all warm-blooded animals, including humans, domesticated and wild animals. Recent studies of Toxoplasma gondii isolates from animals in different regions of China have shown a limited genetic diversity with the dominance of the ToxoDB PCR-RFLP genotype #9 named as “Chinese 1”. However, there is not much published information regarding its prevalence in domestic animals from Guizhou province, a subtropical region in Southwest China. The objectives of this study were to determine seroprevalence and genetic diversity of T .gondii in pigs, dogs and cats in Guizhou province, Southwest China. Findings The anti- T. gondii IgG were detected in 70.0%(49/70) pigs, 20.56%(22/107) dogs and 63.16(12/19) cats. The anti- T. gondii IgM were found in 0.93%(1/107) dogs, 21.53%(4/19) cats, but not in pigs. In addition, the toxoplasma circulating antigen (CAG) were detected in 16.9%18/70)pigs, 13.1% (14/107) dogs and 10.5%(2/19) cats. The T. gondii DNA were detected in 31.5%(22/70) pigs, 3.7%(4/107) dogs and 52.63%(10/19) cats. Five T. gondii isolates were obtained(3 from pigs and 2 from cats). The genotype of these five isolates belonged to the predominant genotype “Chinese 1”. Conclusions The high prevalence of T. gondii infection in pigs,cats and dogs indicated that the T. gondii infection is common in Guizhou province. Additionally, the T. gondii genotype “Chinese 1” was dominant in Southwest China.

Blood samples were collected from patients with leukemia (n = 150) or lymphoma (n = 150) in the Cancer Hospital from March to September 2014. The specific antibodies (IgG, IgM) to, and circulating antigens (CAg) of Toxoplasma gondii were determined by ELISA. A 529 bp specific sequence was amplified by PCR from the genomic DNA of T. gondii. T. gondii-specific IgG positive rate in patients with leukemia and lymphoma were 16.0% (24/150) and 20.0% (30/150), respectively, which were significantly higher than that of healthy persons (6.4%, 7/110) (P < 0.05). IgM positive rate of the leukemia patients, lymphoma patients, and healthy persons was 2.7% (4/150), 1.3% (2/150), and 0.9% (1/110) (P > 0.05), respectively. No significant difference was found in IgM and CAg positive rate among leukemia patients, lymphoma patients, and healthy persons (P > 0.05). No specific band (529 bp) was detected in all samples.

目的:分析弓形虫感染小鼠后Th17细胞及相关细胞因子变化,探讨Th17细胞及相关细胞因子可能的免疫病理作用.方法:Giemsa染色、常规病理学检查、免疫荧光法及PCR扩增弓形虫核酸,确定弓形虫速殖子感染BALB/c小鼠模型的建立;流式细胞术动态检测感染小鼠脾脏中CD3+ CD4+ IL-17+ Th17细胞变化;ELISA法动态检测血清中IL-17、IL-6以及抗弓形虫特异性IgM抗体的变化.结果:感染鼠外周血、腹腔液、肝、脾及肺脏中均可检测弓形虫速殖子,CD3+ CD4+ IL-17+Th17细胞在感染后12 h显著增高,72 h至96 h降低,与正常对照组比较差异均有统计学意义(P＜0.05);小鼠血清中IL-17在感染后12 h至36 h升高,而IL-6则在感染后60 h至72 h先降低,84 h至132 h升高,与对照组比较均有统计学意义(P＜0.05).结论:弓形虫感染小鼠后诱导的Th17细胞及其相关细胞因子可能参与小鼠的免疫病理过程.

Objective: To investigate whether the chromosomes of human umbilical cord mesenchymal stem cells(UC-MSCs) will change after being cultured in vitro for generations.Methods: Umbilical cords of cesarean delivery were collected aseptically.UC-MSCs were isolated by digestion with collagenase and trypsin or mechanical methods,and were cultured in vitro.The cells were characterized by morphology,immunophenotype and cell proliferation test,and karyotypes of generations of P0,P3,P4,P5,and P6 were analyzed with G-banding technique.Results: The karyotype analysis showed a normal diploid karyotype with 46 chromosomes and no abnormal change was found in chromosome structure.UC-MSCs exhibited fibroblastic morphology and CD73,CD90,and CD44 highly expressed in generations P3 ~ P6.Cells reached logarithmic growth phase in 2 ~ 3 days of culture.Conclusion: When cultured for 6 passages in vitro,UC-MSCs can maintain stable chromosome structure,which provides an experimental basis for the safety of UC-MSCs cytotherapy.

Background: Recent studies of Toxoplasma gondii isolates from animals in different regions of China have shown a limited genetic diversity and type China 1 was the dominant genotype of T. gondii prevalent in Chinese animals. However, little has been known concerning the isolation and genotyping of T. gondii circulating in chickens, pigs and rodents in China. The aim of the study was to characterize samples of T. gondii isolates obtained from naturally infected cats, pigs and free-range chickens slaughtered for human consumption in China.Methods: In the present study, brain tissues of 77 animals collected from different areas of China, including 24 free-range chickens (Gallus domesticus), 13 voles (Rattus flavipectus), 23 pigs and 17 cats, were bioassayed in mice and viable T. gondii were isolated from the brains of eleven. These eleven T. gondii isolates were maintained in Kunming (KM) outbred mice and DNA isolated from tissues of infected mice was characterized using 11 PCR-restriction fragment length polymorphism (PCR-RFLP) markers: SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico, and CS3. Moreover, to determine mouse virulence of China 1 lineage of parasites, a TgCtgy5 genotype isolate was selected randomly and assessed in KM mice with different inoculation doses.Results: Results of genotyping revealed that ten isolates were type China 1 (ToxoDB PCR-RFLP genotype #9), and TgCksz1 was a new genotype that was reported for the first time designated here as ToxoDB PCR-RFLP #225. No clonal types I, II and III lineages were found. DNA sequencing of four introns (EF1, HP2, UPRT1 and UPRT7) and two genes (GRA6 and GRA7) from representative isolates confirmed the results of PCR-RFLP genotyping. The TgCtgy5 isolate was highly virulent in KM mice; all infected mice died of acute toxoplasmosis, irrespective of the inoculation dose. The results indicate that mouse virulent isolates of T. gondii are predominantly circulating in cats in China.Conclusions: T. gondii isolated from chickens, pigs, cats and rodents in different locations in China were genotyped and the results reconfirmed the limited diversity of T. gondii in China and showed that type China 1 lineage was dominant in this country.

检验医学专业本科生毕业论文是本科生培养方案中的重要环节之一,通过毕业论文,培养学生综合运用所学知识和技能,提高分析与解决实际问题的能力,在实践中培养学生勇于探索的创新精神,严肃认真的科学态度和严谨求实的工作作风。如何提高毕业论文质量至关重要。贵阳医学院医学检验系自2004年以来,对检验医学本科生毕业论文带教的每个环节制定了严格的要求,使得毕业论文质量大大提高。

Objective:To explore the polarization of Th1 and Th2 immune response and the changes of transcription factor(T-bet and GATA-3) in BALB/c mice infected with toxoplasma gondii.Methods:Sixty BALB/c mice were infected with toxoplasma tachyzoite(1×104/m,0.5 ml) by intraperitoneal injection and thirty BALB/c mice only received an injection of same amount of normal saline as control.After injection,two mice of infection group and one mouse of control group were killed in odd number days.Then,the serum IFN-γ and IL-4 were measured by ELISA,and the mRNA expression of T-bet and GATA-3 in mice spleens were detected by Real-Time RT-PCR.Results:In the infection group of mice,the IFN-γ was remarkable increased at the fourth day and kept the peak from the 5th day to 7th day.Then,it decreased to normal level at 9th day postinjection;but IL-4 was remarkable increased at the 8th day and reached to the peak at 9th day.Then it decreased to normal level at 15th day postinjection.The mRNA of T-bet expression was increased at the third day and kept the peak from the 5th day to 7th day.Then,it decreased to normal level at 9th day postinjection.The mRNA of GATA-3 expression was increased at the 7th day and rised to the peak at 11th day.Then,it decreased to normal level at 13th day postinjection.There were not obvious changes of IFN-γ and IL-4 or T-bet and GATA-3 in normal mice.Conclusion:The Th1 immune response is dominant in acute infection when mice are infected by toxoplasma.Then it biases toward Th2 response at 9th day to 13th day postinfection.The shift of Th1/Th2 polarization influences the finale of toxoplasma infection.

ifferent in various population and regions. This variation could be the cause of the different morbility and clinical characteristics in the coronary heart disease.

[Objective]To study the pathological changes in Kunming mice and Babl/c mice infected with toxo-plasma gondii. [Methods] Kunming mice and Babl/c mice were infected with toxoplasma tachyzoite by intraperi-toneal injection. Then, the infection rate and pathological histology were observed in infected mice. The control mice only received an injection of same amount of saline. [Results]The two kind of mice were susceptible to toxoplasma gondii. The Kunming mice showed infection at 2nd-3rd day postinoculated, and died at 7th～10th day postinfection. The mortality was 95.00%. The hepatocyte denaturation and necrosis were observed. And a lot of toxoplasma tachy-zoites could be obtained from abdominal cavity. But in Babl/c mice, the onset of the sickness was at 4th ～ 5th day postinfection. Most mice could live through the acute stage of infection. The mortality was only 10.00%. The toxo-plasma antigen could be detected in mouse liver and brain, Although the cyst could not be found. All control mice were normal in the research time. [Conclusion] The susceptibility of toxoplasma infection in these two kinds of mice were different. The Babl/c mouse probably is a good model to research the toxoplasma infection and immunity.

Objective: To study the single nucleotide polymorphisms (SNP) in the promoter region of tumor necrosis factor alpha (TNF-α)gene in Han population of Guizhou province. Methods: The five polymorphism sites at -1 031, -863, -857, -308, -238 in TNF-α gene promoter region were detected using PCR-RFLP and SSP-PCR and the data were analyzed statistically. Results: Allele frequencies of the five sites of TNF-α gene promoter region were obtained as the following: -238 G was 96%, -238 A 4%, -308 G 92%, -308 A 8%,-857 C 65%, -857 T 35%, -863 C 80%, -863A 20%, -1 031T 48%, and -1 031C 52% respectively. Conclusions: Unique characteristics exist in the SNP of TNF-α gene promoter region in Han population of Guizhou province. These results provide basic data for the study of the relationship between the polymorphism of TNF-α gene and the diseases.

OBJECTIVE:To investigate the effects of Ginkgo biloba extract (Ginaton) on bcl-2 and bcl-xL mRNA expression in the myocardium of patients underwent hypothermic cardiopulmonary bypass (CPB).METHODS:Thirty congenital heart disease patients were randomly assigned to 2 groups, the control group and the treated group. Patients in both groups received St. Thomas' cardioplegic solution via radix aortae, while Ginaton (0.5 mg/kg) was added in the treated group. Cardiac surgery was started after complete heart arrest. Myocardium was taken before the aorta ascendens was unblocked and mRNA expression of bcl-2 and bcl-xL in the ventricular tissue was detected by RT-PCR.RESULTS:The gene expressions of bcl-2 and bcl-xL were significantly higher in the treated group than those in the control group (P < 0.05).CONCLUSION:Ginaton could promote the mRNA expressions of the antiapoptotic gene bcl-2 and bcl-xL in myocardium of patients underwent CI'PB.