明代《故武略将军致仕副千户薛公墓志铭》《诰封太宜人苗氏墓志铭》考述

山西大同大学档案馆收藏有明代薛茂与苗氏夫妇的两方墓志铭拓片。墓志铭详细记载了薛茂因祖父薛彦中功勋而补官朔州卫百户,先护随明成祖朱棣征讨漠北,后在大同“专领骑队”,全程参战土木之变,之后由于屡立战功,被升为武略将军,授副千户之事。两方墓志铭客观梳理了明太祖朱元璋至明宪宗成化二十年百余年间朔州薛氏族系,还原了薛斌、薛茂、薛端、薛琦、钱能、钱泰、钱宝等人在朔州的史实,对研究明代功勋承袭、官员补荫、官场结交、婚嫁择偶、丧葬立石等社会文化有重要价值。两方墓志铭的撰文、书丹、篆额,皆为当时有名望之士。

苗氏八纲调形手法治疗腰痛理论探析

苗氏八纲调形手法是在天津市宝坻区流传百余年的一种中医传统治疗手法,现已列入天津市宝坻区第3批传统医药类非物质文化遗产代表性项目名录,在长期的临床应用中效果显著。该文以腰痛为例,简要介绍苗氏八纲调形手法的治疗方法,并对其进行了初步的理论溯源与分析,以期为苗氏八纲调形手法的进一步传承、发掘及临床研究提供思路。

科举与仕宦——上党苗氏家族的沉浮

北朝至隋这一时期,苗氏乃是上党的地方大族,苗氏成员大多担任诸如郡县中正、主簿、功曹、县令、县丞等地方官职,家族拥有较为良好的经济状况,通婚对象也是上党大姓,但总体上仍旧属于寂寂无闻的状态。唐朝初年,苗氏家族中的苗琎感知时代的变革,致力于教育子侄,投身科场,在开、天之际,以苗延嗣为首的苗氏成员凭借科举入仕,逐渐从上党地方走向了两京地区,苗氏家族开始了“中央化”和“官僚化”进程。经过苗延嗣、苗晋卿两代人的努力,苗氏家族迈上了一个新台阶,上党苗氏最终从一个地方家族变为“两京衣冠”之一。中晚唐时期,苗氏子弟多凭门荫或科举入仕,并世代与其他官僚家族联姻,苗氏地位得以维持,“中央化”和“官僚化”进程彻底完成。

Immune Effect of Combination of Universal Molecular Adjuvant and IBDV Subunit

In order to evaluate the immune effect of the protein expressed by the universal vector pET-mLTA-CTLA-4 plus IBDV subunit, the fusion protein mLTA-CTLA-4 was expressed and purified. Protein toxicity tests were carried out on rabbits.The VP2 gene of infectious bursal virus was amplified by RT-PCR, and lately used for pET-VP2 construction. Ten-day-old free healthy chickens were chosen for a grouped test, including the mLTA-CTLA-4（at different doses） plus VP2 groups, IBDV living vaccine group and control group. Serum and mucosal samples were collected regularly and the neutralization titers of IgG and IgA were assayed, while an animal protection test was conducted to determine the protection rate. The results showed that the protein m LTA-CTLA-4 was non-toxic and its protection rate was100%. IgG or IgA levels in the IBDV vaccine group were slightly higher than those in recombinant protein groups. These results indicated that the recombinant protein mLTA-CTLA-4 could be applied with IBDV subunit vaccine to protect chickens from infection.

唐代潞州地区墓志的书写格套及演变——兼谈作为移民史史料的误区

唐代潞州地区出土墓志数量众多,多涉及社会中下阶层,墓志大量自云祖先“因官徙家”“某某太守+某某侯”等是一种书写格套,多依据“书仪式”文本拼接而成,具有明显的地域特征,不当视为移民史史料。墓志的书写格套与志盖形制也会因时而变,晚唐五代潞州墓志的书写格套与全国渐趋一致。上党苗氏家族迁至洛阳后,放弃潞州当地墓志简陋的书写格套,完成了祖先叙事的士人化,“上党太守+某某侯”仍时而出现家族记忆中。

Oral attenuated Salmonella typhimurium vaccine against MG7-Ag mimotope of gastric cancer

AIM: To develop an oral attenuated Salmonella typhimurium vaccine against gastric cancer and to evaluate its efficacy in mice. METHODS: A complementary sequence of Nco Ⅰ site and a sequence coding for MG7-Ag mimotope were designed at the 5'terminus of forward primer. Using p1.2 Ⅱ-HBCAg plasmid as template, PCR was performed to get a fusion gene of the mimotope and a HBcAg gene. The fusion gene was then subcloned into the plasmid pYA3341 complementary to Salmonella typhimurium X4550, and the recombinant plasmid was then transformed into attenuated Salmonella typhimurium X4550. Balb/c mice were orally immunized with the recombinant Salmonella typhimurium X4550. The mice were immunized every 2 wk to reinforce the immunity. At the 6th wk, serum titer of antibody was detected by ELISA, and at the 8th wk, cellular immunity was detected by 51Cr release test. Ehrlich ascites carcinoma cells expressing MG7-Ag were used in tumor challenge assay as a model to evaluate the protective effect of the vaccine. RESULTS: Serum titer of antibody against MG7-Ag was significantly higher in mice immunized with the vaccine than in control groups (0.9538±0.043 vs 0.6531±0.018, P<0.01; 0.9538±0.043 vs 0.6915±0.012, P<0.01), while in vitro 51Cr release assay of the splenocytes showed no statistical difference in the three groups. Two weeks after tumor challenge, 1 in 5 immunized mice was tumor free, while all the mice in the control group presented tumor. CONCLUSION: Oral attenuated Salmonella typhimurium vaccine against the MG7-Ag mimotope of gastric cancer is immunogenic. It can induce significant humoral immunity against tumors in mice, and has some protective effects.

Construction of a recombinant attenuated Salmonella typhimurium DNA vaccine carrying Helicobacter pylorihpaA

AIM: To construct a recombinant attenuated Salmonella typhimurium DNA vaccine carrying Helicobacter pylori hpaA gene and to detect its immunogenicity. METHODS: Genomic DNA of the standard H pylori strain 17 874 was isolated as the template, hpaA gene fragment was amplified by polymerase chain reaction (PCR) and cloned into pUCmT vector. DNA sequence of the amplified hpaA gene was assayed, then doned into the eukaryotic expression vector pIRES through enzyme digestion and ligation reactions. The recombinant plasmid was used to transform competent Escherichia coliDH5α, and the positive clones were screened by PCR and restriction enzyme digestion. Then, the recombinant pIRES-hpaA was used to transform LB5000 and the recombinant plasmid isolated from LB5000 was finally used to transform SL7207. After that, the recombinant strain was grown in vitro repeatedly. In order to identify the immunogenicity of the vaccine in vitro, the recombinant pIRES-hpaA was transfected to COS-7 cells using Lipofectamine^(TM)2000, the immunogenicity of expressed HpaA protein was detected with SDS-PAGE and Western blot. RESULTS: The 750-base pair hpaA gene fragment was amplified from the genomic DNA and was consistent with the sequence of H pylori hpaA by sequence analysis. It was confirmed by PCR and restriction enzyme digestion that H pylori hpaA gene was inserted into the eukaryotic expression vector pIRES and a stable recombinant live attenuated Salmonella typhimurium DNA vaccine carrying H pylori hpaA gene was successfully constructed and the specific strip of HpaA expressed by pIRES-hpaA was detected through Western blot. CONCLUSION: The recombinant attenuated Salmonella typhimurium DNA vaccine strain expressing HpaA protein with immunogenicity can be constructed and it may be helpful for further investigating the immune action of DNA vaccine in vivo.

Construction of recombinant attenuated Salmonella typhimurium DNA vaccine expressing H pylori ureB and IL-2

AIM： To construct a recombinant live attenuated Salmonella typhimurium DNA vaccine encoding H pylori ureB gene and mouse IL-2 gene and to detect its immunogenicity in vitro and in vivo. METHODS： Hpylori ureB and mouse IL-2 gene fragments were amplified by polymerase chain reaction （PCR） and cloned into pUCmT vector. DNA sequence of the amplified ureB and IL-2 genes was assayed, then cloned into the eukaryotic expression vector pIRES through enzyme digestion and ligation reactions resulting in pIRES-ureB and pIRES-ureB-IL-2. The recombinant plasmids were used to transform competent E. co/i DH5α, and the positive clones were screened by PCR and restriction enzyme digestion. Then, the recombinant pIRES-ureB and pIRES-ureB-IL-2 were used to transform LB5000 and the recombinant plasmids extracted from LB5000 were finally introduced into the final host SL7207. After that, recombinant strains were grown in vitro repeatedly. In order to detect the immunogenicib/of the vaccine in vitro, pIRES-ureB and pIRES-ureB-IL-2 were transfected to COS-7 cells using LipofectamineTM2000, the immunogenicity of expressed UreB and IL-2 proteins was assayed with SDS-PAGE and Western blot. C57BL/6 mice were orally immunized with 1 × 10^8 recombinant attenuated Salmonella typhimurium DNA vaccine. Four weeks after vaccination, mice were challenged with 1 × 10^7 CFU of live Hpylori SS1. Mice were sacrificed and the stomach was isolated for examination of H pylon 4 wk post-challenge. RESULTS： The 1700 base pair ureB gene fragment amplified from the genomic DNA was consistent with the sequence of H pylori ureB by sequence analysis. The amplified 510 base pair fragment was consistent with the sequence of mouse IL-2 in gene bank. It was confirmed by PCR and restriction enzyme digestion that H pylori ureB and mouse IL-2 genes were inserted into the eukaryotic expression vector pIRES. The experiments in vitro showed that stable recombinant live attenuated Salmonella typhimurium DNA vaccine carrying ureB and IL-2 genes was successfully constructed and the specific strips of UreB and IL-2 expressed by recombinant plasmids were detected through Western blot. Study in vivo showed that the positive rate of rapid urease test of the immunized group including ureB and ureB-IL-2 was 37.5% and 12.5% respectively, and was significantly lower than that （100%） in the control group （P 〈 0.01）. CONCLUSION： Recombinant attenuated Salmonella typhimurium DNA vaccine expressing UreB protein and IL-2 protein with immunogenicity can be constructed. It can protect mice against H pylori infection, which may help the development of a human-use H pylori DNA vaccine.

Immune responses in mice to oral administration of attenuated Salmonella typhimurium expressing Helicobacter pylori urease B subunit by a lavage technique

Objective: To determine whether attenuated Salmonella typhimurium producing Helicobacter pylori (Hp) urease subunit B(UreB)can elicit specific immune responses against Hp in mice tested by a lavage technique. Methods: Attenuated Salmonella typhimurium producing Hp UreB immunized orally Balb/c mice twice at a 3-week interval. After 12 weeks,mice intestinal secretions were obtained without harm by administering a lavage solution intragastrically. The mice intestinal secretions of immune group were also directly washed out after the mice were killed. The antibody responses were evaluated by using serum and intestinal fluid with ELISA assay. Results: The multiple oral immunizations with SL3261/ pTC01-UreB induced significantly Hp-specific mucosal IgA response as well as serum IgG response. The IgA was also consistently higher in the intestinal fluid obtained by the lavage solution than by direct washout. In addition, no obvious side effects and changes in gastric inflammation were observed in mice. Conclusion: The attenuated Salmonella typhimurium expressing Hp UreB may be used as an oral vaccine against Hp infection. And the lavage technique is an ideal method in the study of mucosal immune responses.

Gene cloning and prokaryotic expression of recombinant outer membrane protein from Vibrio parahaemolyticus

Gram-negative Vibrio parahaemolyticus is a common pathogen in humans and marine animals, The outer membrane protein of bacteria plays an important role in the infection and pathogenicity to the host. Thus, the outer membrane proteins are an ideal target for vaccines. We amplified a complete outer membrane protein gene （ompW） from E parahaemolyticus ATCC 17802. We then cloned and expressed the gene into Escherichia coli BL21 （DE3） cells. The gene coded for a protein that was 42.78 kDa. We purified the protein using Ni-NTA affinity chromatography and Anti-His antibody Western blotting, respectively. Our results provide a basis for future application of the OmpW protein as a vaccine candidate against infection by E parahaemolyticus. In addition, the purified OmpW protein can be used for further functional and structural studies.

Titration Method Validation of Live Vaccines against Infectious Bursal Disease

Validating a method of analysis goes through different steps, which aims at testing the normality of measurements distribution, estimating the uncertainty of the components of a measurement （i.e., accuracy and correctness）, and finally, define the control tests of non degradation of the method performances. This paper outlines the steps for validating a biological method of analysis. It involves the construction of an experimental design, a statistical model, and the preparation of an interne laboratory reference material （pilot vaccine）. The latter is used to study the impact of deviation and variation factors, in order to, optimize the analytical method, to evaluate the bias （random error）, and to calculate the uncertainty of measurement, and make the control charts. This method is applied in the titration of live viral vaccines of Gumboro disease on chicken＇s embryos fibroblasts. The experimental results show that potential influence factors related to the titration method had no significant influence on the obtained results. Taking into account these results, an operating mode has been elaborated. The finalized method proved to be faithful to standard deviation of repeatability and reproducibility of 0.21 and 0.22, respectively, with a confidence level of 95%. The calculated uncertainty of measurement is equal to 0.2, which represents the average error level of a titer. A homogeneous stock of interne laboratory reference vaccine （MRIL）, with an average titer of 5.9 log DIT 50, was produced and the control chart set in away to provide the laboratory with an important tool of control and monitoring of the viral titers evolution in time, as well as, the mastery of the validated titration method performances.

小儿血清苗勒抑制物质水平变化及其临床意义

通过测定非泌尿生殖系统疾病及隐睾患者的血清苗勒抑制物质 (MIS)值 ,观察年龄与MIS的相关性。结果男性非泌尿生殖系统疾病患儿血清MIS值随年龄增长而下降 ,女性儿童血清MIS值明显低于男性 ,且不随年龄变化 ,青春发育前期隐睾儿童血清的MIS值高于同龄儿。

Primary omental malignant mixed Müllerian tumor in a 67-year-old woman

背景 大网膜原发苗勒氏管混合瘤很少见 ,迄今为止仅报道了 2例 ,具有其特殊临床特征。病例 女性 ,6 7岁 ,G6P5 ,主因腹痛、腹胀 15天就诊于北京大学人民医院妇科。体格检查发现一直径 12厘米腹部包块 ,活动度差。行肿瘤及大网膜大部分切除 ,全子宫加双附件切除术。术后病理示大网膜原发恶性苗勒氏管混合瘤。术后顺铂、吡喃阿霉素、足叶已甙化疗一疗程 ,因严重骨髓抑制 ,未继续化疗 ,术后 8个月发现肿瘤肝转移。结论 原发性恶性苗勒氏管混合瘤预后差 ,现今手术仍是最有效的治疗手段 ,化疗及放疗效果不确切。

Effect of electroacupuncture on anti-Mullerian hormone expression in rats with polycystic ovarian syndrome

Objective: To observe whether the effect of electroacupuncture (EA) on improving sex hormone disorders and follicle development is by decreasing the expression of anti-Mullerian hormone (AMH) in rats with experimental polycystic ovarian syndrome (PCOS). Methods: Forty rats were randomly divided into four groups, a normal group (NG), a model group (MG), an EA at acupoints group (EAAG), and an EA at non-acupoints group (EANAG), with 10 rats in each group. The rats in the EAAG and EANAG were intervened by EA treatment for consecutive 14 d. Zhongji (CV 3) and Guanyuan (CV 4) were selected as the acupoints in the EAAG, and the tip of the tail and 1 cm up from the tail tip were selected as the non-acupoints in the EANAG. After treatment, the histomorphological changes of the ovary, the levels of aromatase P450 (P450arom)/ testosterone and estradiol in the ovarian tissues, and the expressions of follicle stimulating hormone (FSH) and AMH were observed. Results: After treatment, compared with the MG and EANAG, the expression of AMH decreased (P<0.05), the levels of P450arom and estradiol in creased sign ifica ntly, and the level of testostero ne decreased sign ifica ntly (all P<0.01) in the EAAG. Additionally, several normal follicles were present and the number of cystically dilated follicles decreased in the EAAG. Compared with the MG and EANAG, the EAAG obviously had more follicular granulosa cells. Con elusion: EA can dow n-regulate the abn ormally in creased expressi on of AMH to improve sex hormone disorders and follicle development in PCOS rats.

Identification of immunoreactive proteins of Brucella melitensis by immunoproteomics

Infection with Brucella causes brucellosis, a chronic disease in humans, which induces abortion and sterility in livestock. Among the different Brucella species, Brucella melitensis is considered the most virulent and is the predominant species associated with outbreaks in China. To date, no safe human vaccine is available against Brucella infection. The currently used live vaccines against Brucella in livestock induce antibodies that interfere with the diagnosis of field infection in vaccinated ani- mals, which is harmful to eradication programs. However, there is as yet no complete profile of immunogenic proteins of B. melitensis. Towards the development of a safer, equally efficacious, and field infection-distinguishable vaccine, we used immunoproteomics to identify novel candidate immunogenic proteins from B. melitensis M5. Eighty-eight immunoreactive protein spots from B. melitensis M5 were identified by Western blotting and were assigned to sixty-one proteins by mass spectrometry, including many new immunoreactive proteins such as elongation factor G, FOFI ATP synthase subunit beta, and OMPI. These provide many candidate immunoreactive proteins for vaccine development.