Objective Choose an easy method to screen the “leaky" phenotype SCID mice before experimental use. Methods IgG in SCID mice serum was assayed by three kinds of ELISA(sandwich ELISA, sandwich indirect ELISA and Avidin\|biotin sandwich ELISA). Results Avidin\|biotin sandwich ELISA is the most sensitive method of the three, and 41 SCID mice of 6～8 weeks old were assayed with this method. The average IgG of 40 SCID mice was 0 24?0 16mg/L, only one was 2\^3mg/L. The “Leaky" probability was 2\^4%.Conclusion\ The Avidin\|Biotin sandwich ELISA is an easy, sensitive and reliable method to screen“leaky" SCID mice.\;[

Objective To determine whether p38 mitogen-activated protein kinase (p38MAPK) signaling pathway is involved in cerebral fractalkine-induced hyperalgesia in mice.Methods Two hundred and twenty-five male Kunming mice weighing 30-40 g were randomly divided into 4 groups:group control ( group C,n =55 ) ;group fractalkine (group F,n =60); group anti-CX3CR1 + fractalkine (group CF,n =55) and group SB203580 (p38MAPK inhibitor) + fractalkine (group SF,n =55).Fractalkine 100 ng was injected into cerebral lateral ventricle (i.c.v.) in groups F,CF and SF.Anti-CX3CR1 1 μg and SB203580 1 μg were injected i.c.v.at 1 h before fractalkine injection in groups CF and SF respectively.Paw withdrawal latency to a thermal nociceptive stimulus (PWL) was measured at 30 min before the drugs were injected into cerebral lateral ventricle and 30,60,120 and 240 min after fractalkine injection.Five animals were sacrificed after PWL measurement at each time point and their brains were removed for determination of phosphorylated p38MAPK protein expression (by Western blot analysis).Five animals were sacrificed at 30 min before the drugs were injected into cerebral lateral ventricle and 6,12 and 24 h after fractalkine injection for determination of IL-1β and TNF-α contents in the brain (by ELISA) in all the 4 groups.In group F 5 animals were sacrificed at 4 h after fractalkine injection for determination of action of fractalkine on microglia or astrocyte (by immunofluorescence).Results Fractalkine i.c.v.injection significantly reduced PWL and increased phosphorylated 38MAPK,IL-1β and TNF-α levels in group F as compared with group C.Pretreatment with anti-CX3CR1 or SB203580 significantly decreased fractalkine-induced hyperalgesia and phosphorylated-p38MAPK,IL-1β and TNF-α levels in groups CF and SF as compared with group F.Fractalkine was localized at microglia.Conclusion p38MAPK signal transduction pathway is involved in cerebral fractalkine-induced hyperalgesia in mice.

Background and purpose: Primary signet ring cell carcinoma(SRCC) of the bladder is rarely diagnosed in the clinic. Few cases have been reported in the literature, so there was lack of understanding of the primary bladder SRCC in terms of diagnosis and treatment. Our study was to investigate the clinical features and treatment strategy for primary SRCC of the bladder and review the status of the disease along with the literature. Methods: 3 cases of primary bladder SRCC were studied, including clinical features, treatment, follow-up and their prognosis.The literature was reviewed. Results: All cases received ultrasound, computerized tomography, cystoscopy, biopsy and other related lab tests for diagnosis and differential diagnosis. Laparoscopic radical cystectomy and orthotopic ileal neobladders were performed in 2 cases, while the other case received laparoscopic radical cystectomy and ileal conduit diversion, Chemotherapy (cisplatin and 5-fluorouracil) was delivered in one case after surgery. One patient died at 6 months postoperatively because of multiple metastasis. The other 2 cases have been followed-up only for 8 and 12 months postoperatively, and no recurrence or metastasis have been observed. Conclusion: Primary SRCC of the bladder lacks distinctive clinical and imaging manifestations. The tumor grows very invasively. Radical cystcctomy is one of the optimal approaches for treatment of SRCC of bladder.

Background and purpose: The prognostic factors on survival for the patients with prostate carcinoma are still underdeterrnined. This study was to analyze the survival of three common treatment methods for prostate carcinoma and the prognostic factors on survival. Methods: 494 male patients who were diagnosed as prostate cancer were enrolled into the retrospective study. All of the data like age, stage, grade, PSA level, ALP, Hb and treatments were collected. Overall survival and disease specific survival rates for patients were analyzed by Kaplan-Meier method. Prognostic factors on disease specific survival were also analyzed by Log-rank test and Cox proportional hazards model. Results: Disease specific survival rates at 1, 3 and 5 year were 96.0%, 89.0% and 80.0% for all 494 patients, respectively. Disease specific survival rate at 3-year was 92.4% for brachytherapy, 100.0% for radical prostatectomy and 80.6% for hormonal therapy (P=0.008). Multivariate analysis by Cox model showed that stage, PSA level and age significantly impacted on disease specific survival. Conclusion: Brachytherapy and radical prostatectomy provides longer survival time than hormonal therapy for patients with prostate cancer. Clinical stage and PSA level and age of prostate cancer are independent factors impacting on survival significantly.

Objective To set up a new diagnostic platform based on microarray exon-capture and next-generation sequencing for detecting small mutations in dystrophin gene.The sensitivity and specificity of the method were assessed in clinical settings and the distribution of small mutations in Chinese Duchenne muscular dystrophy/Becker muscular dystrophy (DMD/BMD) patients were also analyzed.Methods Forty-one DMD/BMD patients diagnosed by the clinical criteria without large deletion or duplication (≥ 1exon) were recruited from Peking Union Medical College Hospital consecutively.Genomic DNA was extracted from blood samples.The libraries were prepared.Then exon and intron-exon flanking sequences of DMD gene were captured by custom microarray.Targeted next-generation sequencing and Sanger Sequencing were conducted.The patients who were not detected any disease-causing mutation were performed muscle biopsy.Results Thirty-eight subjects were detected small mutations in DMD gene.All single nucleotide variants (SNVs) and insertion & deletions (INDELs) were validated by Sanger sequencing.Twenty-one novel mutations were reported.The distribution of SNVs and INDELs was similar to other international DMD databases.Upon immunohistochemistry staining of dystrophin protein,1 of 3 mutation-undetected patients was diagnosed as DMD,2 of them were excluded.The specificity of the method was 100％,while the sensitivity was 97.4％.Conclusions Our microarray-captured next-generation sequencing assay could detect SNVs and INDELs with high sensitivity and specificity.Its advantages are economic,time-saving and stable.The platform is suitable for clinical gene diagnosis.

Objective To investigate the association between extramural vascular invasion (EMVI) detected by multi-detectors computed tomography (MDCT) with contrast enhanced (ceMDCT) and clinicopathologic characteristics in patients with colon cancer.Methods Between February 2009 and December 2013,patients with histologically proven primary colon cancer and undergoing curative resection were included in this retrospective study.According to American Joint Committee on Cancer TNM staging system,patients of stage Ⅱ and Ⅲ were included in this study.EMVI status detected by MDCT (ctEMVI) was defined according to the EMVI scores.Chi-square test was used to analyze the association between clinicopathologic characteristics and ctEMVI.Results 165 stage Ⅱ and stage Ⅲ patients were included in this study as confirmed by pathology based on AJCC.Positive ctEMVI was demonstrated in 51 patients (34.5％,51/165).There were significant association between positive ctEMVI and age ＜ 65 years (x2 =4.810,P =0.031),ceMDCT defined tumor stage (x2 =17.911,P =0.000),ceMDCT defined metastatic lymph node (x2 =5.436,P =0.022),tumor size≥5 cm (x2 =3.799,P =0.036) and pathological T stage (x2 =13.346,P =0.001).Conclusions EMVI,detected by ceMDCT,is significantly associated with age,tumor size and T staging in colon cancer.

Lymphocyte function associated antigen-1 (LFA-1) is a member of integrin family, that plays an important role in the adhesion of lymphocytes with other cells and matrix. To investigate the role of LFA-1 in collagen-induced arthritis (CIA), the incidence of CIA, histological and radiological assessments in the LFA-1 deficient (LFA-1~ -/- ) mice and control mice were examined. LFA-1~ -/- mice and control mice were immunized with 100?g collagen type II(CII) emulsified with an equal volume of Freund’s complete adjuvant (CFA), followed by the booster injection of the same amount of CII in CFA on day 21. Then, clinical, histological and radiological assessments were done. It showed that 57% control mice developed arthritis and apparently changed in the histological and radiological assessment, whereas the all of LFA-1~ -/- mice had the normal histological and radiographic response and none developed arthritis. These results suggeste that LFA-1 is indispensable for the onset of CIA.

Objective:To observe the effects of herbal cake.partitioned moxibustion and bran-partition moxibustion in improving symptoms of ulcerative colitis(UC)and the TNF-α and its receptor of colon mucosa.Method:67 UC cases were randomly allocated into herbal cake-partition moxibustion group of 35 cases and bran-partitioned moxibustion group of 32 cases,to compare the improvement and detect the TNF-α and its receptor with inlmunohistochemical method in both groups.Result:Herbal cake.partitioned moxibustion iS prior to bran-partitioned moxibustion in improving of diarrhea,flatus,lassitude,tenesmus and lumbar soreness;The expression of TNF-α,TNF-αR1,and TNF-αR2 are significantly decreased after treatment in herbal cake-partitioned moxibustion group,while in bran-partitioned moxibustion group only TNF-αR1 expression is significant decreased after treatment.Conclusion:Moxibustion can well improve the syndromes of UC.Herbal cake.partitioned Moxibustion is prior to bran-partitioned moxibustion in the improvement of diarrhea and flatus;Herbal cake-partitioned moxibustion could down-regulate the expression of TNF-α,TNF-αR1.and TNF-αR2.while bran-partitioned moxibustion could only down-regulate the expression of TNF-αR1.

BACKGROUND:The production and release of a large amount of inflammatory factors caused by immune system inflammatory response mainly contributes to secondary spinal cord injury. OBJECTIVE:To investigate the effects of umbilical cord Wharton’s jely mesenchymal stem cel transplantation on repair of injured neurological function and expression of inflammatory factors monocyte chemoattractant protein 1 and interleukin 10 in rats with acute spinal cord injury. METHODS: Eighty-one healthy adult male Sprague-Dawley rats were randomly and equaly divided into sham operation, model and cel transplantation groups, with 27 rats per group. Rats in the latter two groups were subjected to hemisection of the spinal cord to establish acute spinal cord injury models. Rat models in the cel transplantation group received umbilical cord Wharton’s jely mesenchymal stem cel injection (1×106)via the tail vein. Rat neurological function was evaluated using the BBB score at different time points after spinal cord injury. The expression of monocyte chemoattractant protein 1 and interleukin 10 in injured spinal cord tissue was detected using ELISA assay at different time points after spinal cord injury. Migration and neuronal differentiation of umbilical cord Wharton’s jely mesenchymal stem cels in the injured spinal cord tissue were determined using immunohistochemical staining method. RESULTS AND CONCLUSION:Compared with the sham operation and model groups, rat neurological function was significantly recovered in the cel transplantation group (P < 0.05). Compared to the model group, monocyte chemoattractant protein 1 level in the serum and monocyte chemoattractant protein 1 mRNA and protein expression in the injured spinal cord tissue were significantly lower (P < 0.05), but interleukin 10 mRNA and protein expression in the injured spinal cord tissue was significantly higher (P < 0.05), in the cel transplantation group. In the cel transplantation group, umbilical cord Wharton’s jely mesenchymal stem cels could migrate to the injured region and express glial fibrilary acidic protein. These findings suggest that umbilical cord Wharton’s jely mesenchymal stem cels promote rat neurological function recovery by regulating the inflammatory response in the injured spinal cord tissue, which is likely to be one of mechanisms by which transplantation of umbilical cord Wharton’s jely mesenchymal stem cels treats spinal cord injury.

Objective To study the mechanism of marcosomia by investigating insulin-like growth factor 2(IGF_2)imprinting status,expression level and the promoter usage in the placenta of macrosomia. Methods We selected heterozygous cases for Apa Ⅰ polymorphism in exon 9 of IGF_2 gene and then analyzed its imprinting status in 168 placentas of macrosomia and normal pregnancies.IGF_2 transcription levels and promoter usages in macrosomic and normal placenta were evaluated by using semi-quantitative RT- PCR assay.Results Thirty specimens of macrosomic placenta and 30 of normal placenta were identified as heterozygous for IGF_2.All of the heterozygous specimens showed maintenance of imprinting.The expression of placental IGF_2 mRNA(2.2?1.2)was significantly higher in macrosomia than that of normal weight group (1.6?0.6,P 0.05).Conclusion It is possible that over expression of IGF_2 in placenta contributes to macrosomia while the promoter usage and imprinting status are not associated with macrosomia.