This study was designed to explore the RNA interference technique in inhibition of the expression of the mouse fibrinogen like protein 2 (mfgl2), which has been reported to be involved in the development a variety of diseases including fulminant viral hepatitis. A plasmid named p-mfgl2shRNA,complementary to the sequence of mfgl2 was constructed, while another short hairpin RNA (shRNA)which was a mutated form of the mfgl2shRNA sequences was used as a control. A plasmid named pEGFP-mfgl2 expressing the mfgl2-EGFP fusion protein was also constructed for the screening of the effect of p-mfgl2shRNA on mfgl2 expression. By cotransfection of p-mfgl2shRNA and pEGFP-mfgl2 or pcDNA3.1-mfgl2 expression construct into CHO cells or HeLa cells, the inhibition of mfgl2 expression by mfgl2shRNA was analyzed by direct observation through fluorescent microscopy, FACS, RT-PCR and immunohistochemistry staining. The experiments showed the significant inhibitory effect of p-mfgl2shRNA on mfgl2 expression at 48h post-transfection in both CHO and Hela cell lines with the inhibitory efficiency as high as 80.1%. The study demonstrated that the construct of p-mfgl2shRNA successfully interfered with the mfgl2 expression in vitro.

To study the contribution of T cell subsets in the pathogenesis of Murine hepatitis virus Type3 (MHV-3) induced chronic viral hepatitis in C3H/Hej mice, ninety C3H/Hej mice were chosen to individually receive 10 plaque forming units (PFU) of MHV-3 intraperitoneally. The changes of virus titer and pathology in liver tissue were examined by standard plaque assay and by the hematoxylin/eosin (HE) staining method from 2 days post MHV-3 infection. The ratios of T cell subsets including CD3+CD4+CD8-, CD3+CD4-CD8+, CD3+CD4-CD8-, CD3+CD4+CD25+, CD3+CD4+CD25- and CD3+CD4-CD25+ T lymphocyte of total T lymphocytes in blood, spleen and liver were examined at 0, 2, 4, 6,8, 10, 12, 15, 20, 25, 30, 40 days post MHV-3 infection by flow cytosorting. We observed that the virus titer raised and showed persistent virus duplications and inflammatory changes in the livers of C3H/Hej mice from 2 days post MHV-3 infection. The double negative T cell (DN Treg cell) and CD4+CD25+ T cell ratios increased significantly from 2 days post MHV-3 infection in C3H/Hej mice, and CD3+CD4+CD8-, CD3+CD4-CD8+, CD3+CD4+CD25- and CD3+CD4-CD25+ T cell ratios decreased accordingly. In conclusion, the changes of virus titer and pathology in the livers of C3H/Hej mice post MHV-3 suggest their contribution to viral persistence. Further characterizations of DN Treg cells are that infection indicates that MHV-3 could induce the chronic inflammation in livers of C3H/Hej mice.The increase of the DN Treg cell and CD4+CD25+ T cell ratios in C3H/Hej mice post MHV-3 infection suggests that DN Treg cells and CD4+CD25+ T cells may both have important suppressive immunomodulation functions in the development of chronic viral hepatitis and have important roles in the virus persistent infection. Further characterizations of DNT cell and CD4+CD25+ T cell are under investigation.

Abstract: Mycobacterium senegalense is one of the major pathogens causing bovine farcy, and reports of its infection in human are rare. Here is a report on a woman who had been taking hormones and immunosuppressants for a long time for SLE and underwent abdominal soft tissue infection with Mycobacterium senegalense after abdominal liposuction, to provide reference for clinical diagnosis and treatment. The patient, female, 32 years old, has a history of SLE for more than 2 years, and currently takes "methylprednisolone, hydroxychloroquine, and mycophenolate mofetil" regularly. Nine months before the patient was admitted to the hospital, she once performed abdominal, waist and buttock liposuction in a medical beauty institution. One month after the operation, several masses gradually appeared on the abdominal wall, accompanied by tenderness, one of the masses had obvious fluctuation on palpation and purulent fluid could be drawn out. The location of the abdominal wall mass was consistent with the insertion site of the liposuction needle. After the onset of the disease, the patient went to the medical beauty institution for puncture of the abdominal wall mass, and 5 mL of purulent fluid was pierced and sent for bacterial culture, and cultured "Mycobacterium Senegalense", after 3 days of treatment with "cephalosporin" antibiotics (specifically unknown), the symptoms did not improve, so she went to the second affiliated hospital of hainan medical college. After completing the relevant examinations during the hospitalization in our hospital, in order to clarify the etiology, another abdominal puncture to extract pus was performed, the mycobacterial culture + identification results: Mycobacterium senegalense. Consistent with the out-of-hospital results, the diagnosis of Mycobacterium senegalense infection was confirmed. After 3 months of treatment with "cefoxitin, azithromycin, amikacin, and levofloxacin", the patient's abdominal wall soft tissue infection was cured. Trauma or invasive procedures can lead to skin, muscle, or bone infection with nontuberculous mycobacteria (NTM), which can manifest as chronic painless nodules that progress to purulent folliculitis and abscesses. NTM infection should be suspected when the patient's wound has been exposed to water, there is a history of surgery, and empirical anti-infection is ineffective. This is the first case of Mycobacterium senegalense infection caused by medical beauty, which tell people that they should be cautious when choosing medical aesthetic projects and medical aesthetic institutions.

Department of Pediatrics, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China. zhuchuanlong@jsph.org.cn.
Adolescent ; Child ; Female ; Glycyrrhizic Acid ; therapeutic use ; Humans ; Male ; Non-alcoholic Fatty Liver Disease ; drug therapy ; Tablets

Adult ; Aged ; Cell Cycle Proteins ; analysis ; Coal Mining ; Cyclin-Dependent Kinase Inhibitor p27 ; Female ; Humans ; Immunohistochemistry ; Lung ; chemistry ; pathology ; Male ; Middle Aged ; Occupational Diseases ; metabolism ; Pneumoconiosis ; metabolism ; Tumor Suppressor Proteins ; analysis

OBJECTIVETo evaluate the clinical practice of laparoscopic rectopexy in the treatment of severe rectal prolapse.

METHODSFrom March 1998 to February 2007, 4 cases of complete rectal prolapse, including 1 male and 3 female,ranged 21-82 years old, were treated by laparoscopic rectopexy. In one case, the posterior wall of rectum was freed and elevated, and pre-rectal introcession was closed by silk suture, then the posterior wall was suspended and fixed on sacral promontory fascia, finally the sigmoid colon was fixed by sutures on the fascia of left psoas major. In other three cases, insertion of mesh was performed. Rectum was freed and elevated to the level of levalor ani. A sheet of T-shape polypropylene mesh was placed posterior to the rectum, whose lower margin was at the level of levator ani and wrapped around the rectum covering except the anterior wall. The free margin of the mesh was sutured on the muscular layer of rectum, then the mesh was put posterior to the rectum and fixed on the sacral promontory fascia by clipping to repair hernia. After that, the pelvic peritoneum was closed, and finally the sigmoid colon was fixed by sutures on the fascia of left psoas major.

RESULTSFour operation procedures were completed successfully. There was no conversion operation. The time was consumed 92.5 (80-100) min, and the bleeding amount was 6.5 (5-10) ml. No post-operative complications were found. Urine incontinence and encopresis were relieved. No recurrence and constipation was found after 2 months to 3 years follow up postoperatively.

CONCLUSIONLaparoscopic rectopexy is a safe, workable and effective procedure, which can reduce operative trauma and shorten hospitalization time.

Aged ; Aged, 80 and over ; Female ; Humans ; Laparoscopy ; Male ; Rectal Prolapse ; surgery ; Rectum ; surgery ; Young Adult

OBJECTIVETo examine serum tissue inhibitors of metalloproteinases (TIMP) -1 and -2 levels in children with nonalcoholic fatty liver disease and to investigate possible roles of the two markers.

METHODSOne hundred and five obese children were classified into 4 groups: simple obesity (n=44), simple nonalcoholic fatty liver (SNAFL, n=25), and nonalcoholic steatohepatitis (NASH, n=36). Serum TIMP-1 and -2 levels were measured using ELISA. Serum ALT and gamma-GT levels were measured with totally automatic enzymatic method.

RESULTSSerum levels of TIMP-1 and gamma-GT increased with the disease development from simple obesity to SNAFL and NASH (P<0.05). Both serum TIMP-1 and -2 levels were positively correlated with gamma-GT levels (r=0.534, P<0.01; r=0.351, P<0.05, respectively). Ninety-seven percent of children in the NASH group had serum TIMP-1 levels over 2 standard deviations of healthy controls (83.35 microg/ L) compared with 76% in the SNAFL group (P<0.05). There were no significant differences in the case proportion with TIMP-2 levels over 2 standard deviations of healthy controls between the NASH and the SNAFL groups.

CONCLUSIONSBoth TIMP-1 and -2 may reflect the state of liver fibrosis in children with nonalcoholic fatty liver disease, and serum TIMP-1 appears to be more reliable.

Adolescent ; Alanine Transaminase ; blood ; Child ; Enzyme-Linked Immunosorbent Assay ; Fatty Liver ; blood ; Female ; Humans ; Male ; Tissue Inhibitor of Metalloproteinase-1 ; blood ; Tissue Inhibitor of Metalloproteinase-2 ; blood ; gamma-Glutamyltransferase ; blood

This study was aimed to analyze the polymorphism of HLA-A, B, DRB1 alleles at high-resolution level in Han population from southern area of Shandong province in China. 688 randomly selected, unrelated and healthy individual from southern area of Shandong province were genotyped for HLA-A, -B and HLA-DRB1 loci by PCR-SBT. Then, allelic and haplotypic distributions of HLA-A, B and DRB1 were estimated by maximum likelihood estimation method using Arlequin 3.0. The results indicated that a total of 31 HLA-A, 63 HLA-B and 39 HLA-DRB1 alleles were identified in Han Population from southern area of Shandong province. Six HLA-A alleles were found with a frequency greater than 0.05 (A*24:02, *30:01, *11:01, *02:01, *33:03 and *02:06), with a cumulative frequency of 0.7223. For HLA-B locus, there were also six alleles which had a frequency higher than 5% (B*1302, *4403, *5101, B*4601, *1501 and *5801), representing 0.4432 of the all alleles in the population. And four HLA-DRB1 alleles were defined as predominant (DRB1*0701, *1501, *0901and *0803), accounting for 0.5453 of the defined alleles. The most common three-loci haplotype was A*30:01-B*13:02-DRB1*07:01 (0.1151) and the most frequent two-loci haplotype were A*30:01-B*13:02 (0.1303), A*30:01-DRB1*07:01 (0.1157) and B*13:02-DRB1*07:01 (0.1307). It is concluded that the allelic and haplotypic diversities of HLA-A, -B and HLA-DRB1 at high-resolution in Han population from southern area of Shandong province in China provide useful information for HLA matching in transplantation and diseases-associated study in this population.
Alleles ; Asian Continental Ancestry Group ; genetics ; China ; Female ; Gene Frequency ; Genetics, Population ; HLA-A Antigens ; genetics ; HLA-B Antigens ; genetics ; HLA-DRB1 Chains ; genetics ; Haplotypes ; Humans ; Male ; Polymorphism, Genetic

OBJECTIVETo identify a novel human leukocyte antigen (HLA) allele in Chinese and investigate its inheritance in the family.

METHODSExceptional reaction pattern was detected in HLA-B locus in HLA typing using Luminex DNA polymerase chain reaction with sequence specific oligonucleotide probe hybridization (PCR-SSOP) assay. A confirmatory test for the novel HLA allele was performed by DNA sequencing based typing of the proband's family.

RESULTSThe DNA sequence was confirmed to be a novel HLA B allele. There were 7 nucleotides which differed from the closest matching HLA B*40:06:01 at positions 302(G to A), 309(G to C), 311(A to C), 313(C to G), 314(T to C), 317(G to T), and 319(G to C) in exon 2, which resulted in 5 amino acid changes at codon 101 (Ser to Asn), 104 (Asn to Thr), 105 (Leu to Ala), 106 (Arg to Leu), and 107 (Gly to Arg), respectively. Family investigation indicated that the novel allele was transmitted from the proband's father.

CONCLUSIONA novel HLA B allele was identified and officially named as HLA-B*40:96 (GenBank accession No. FJ374890) by the WHO Nomenclature Committee for Factors of the HLA System.

Alleles ; Base Sequence ; Female ; HLA-B Antigens ; genetics ; Haplotypes ; Histocompatibility Testing ; Humans ; Molecular Sequence Data ; Pedigree ; Sequence Alignment

AIMTo assess the psychometric properties of the Chinese Index of Premature Ejaculation (CIPE).

METHODSThe sexual function of 167 patients with and 114 normal controls without premature ejaculation (PE) were evaluated with CIPE. All subjects were married and had regular sexual activity. The CIPE has 10 questions, focusing on libido, erectile function, ejaculatory latency, sexual satisfaction and difficulty in delaying ejaculation, self-confidence and depression. Each question was responded to on a 5 point Likert-type scale. The individual question score and the total scale score were analyzed between the two groups.

RESULTSThere were no significant differences between the age, duration of marriage and educational level (P> 0.05) of patients with and without PE and normal controls. The mean latency of patients with PE and normal controls were 1.6 +/- 1.2 and 10.2 +/- 9.5 minutes, respectively. Significant differences between patients with (26.7 +/- 4.6) PE and normal controls (41.9 +/- 4.0) were observed on the total score of CIPE (P< 0.01). Using binary logistic regression analysis, PE was significantly related to five questions of the original measure. They are the so-called the CIPE-5 and include: ejaculatory latency, sexual satisfaction of patients and sexual partner, difficulty in delaying ejaculation, anxiety and depression. Receiver Operating Characteristic (ROC) curve analysis of CIPE-5 questionnaire indicated that the sensitivity and specificity of CIPE were 97.60 % and 94.74 %, respectively. Employing the total score of CIPE-5, patients with PE could be divided into three groups: mild (>15 point) 19.8 %, moderate (10-14 point) 62.8 % and severe (< 9 point) 16.7 %.

CONCLUSIONThe CIPE-5 is a useful method for the evaluation of sexual function of patients with PE and can be used as a clinical endpoint for clinical trials studying the efficacy of pharmacological intervention.

Adult ; Aged ; Algorithms ; China ; Coitus ; Ejaculation ; Humans ; Logistic Models ; Male ; Middle Aged ; Orgasm ; physiology ; Penile Erection ; physiology ; Psychometrics ; ROC Curve ; Sexual Dysfunction, Physiological ; classification ; diagnosis ; psychology ; Surveys and Questionnaires ; Time Factors