BACKGROUNDThe heavy incidence and mortality of hemorrhagic fever with renal syndrome, as well as no specific drugs in curing the disease, clearly indicate the need for development of the more effective hantavirus vaccine. Refining the DNA vaccination strategy to elicit more clinically efficacious immune responses is now under intensive investigation. In the present study, we examined the effects of using an interleukin-12 expression plasmid as a genetic adjuvant to enhance the immune responses induced by a DNA vaccine based on the S gene encoding nucleocapsid protein against hantavirus.

METHODSBALB/c mice were immunized three times by intramuscular inoculations of DNA vaccine encoding of hantanvirus nucleocapsid protein alone or in combination with a plasmid expressing murine interleukin-12 (pcIL-12). Booster immunizations were employed 2 times at 2-week interval. To evaluate the humoral and cellular immune responses, antigen-specific lymphocyte proliferation and antibody production were assayed by MTT method and ELISA respectively. The level of interleukin-4 and interferon-gamma in the splenic lymphocytic cultured supernatant were detected with ELISA kit at day 5, 10, 17, 35 and 42 after primary immunization.

RESULTSAntigen-specific IgG antibodies was increased markedly at day 17 in the experiment groups and reached a plateau after day 35. As pcIL-12 co-injected, a significant inhibition of antigen-specific IgG levels was displayed over the period and the antibody mean titre was decreased to only about 1:50 at day 42 after primary immunization, significantly lower than the group immunized with pcDNA3.1 + S alone, in which the mean titre was about 1:70. Interferon-gamma was increased remarkably by the co-injection of pcIL-12 compared with the injection of pcDNA3.1 + S alone. However, the production of interleukin-4 was inhibited by pcIL-12 co-injection. Furthermore, pcIL-12 co-injection efficiently enhanced antigen-specific lymphocyte proliferation.

CONCLUSIONHumoral and cytokine responses elicited by pcDNA3.1 + S inoculation can be modulated by co-inoculation with pcIL-12 and efficiently induced Th1-dominant immune responses.

Animals ; Cytokines ; biosynthesis ; Genetic Therapy ; Hantavirus ; immunology ; Immunoglobulin G ; blood ; Immunophenotyping ; Interleukin-12 ; genetics ; Lymphocyte Activation ; Male ; Mice ; Mice, Inbred BALB C ; Nucleocapsid ; genetics ; immunology ; Vaccines, DNA ; immunology ; Viral Vaccines ; immunology

OBJECTIVETo study the difference of gene expression profiles in gastric cancer (T), pericancerous mucosa (P) and the gastric mucosa from distant cutting margin (C), and to screen an associated novel gene in early gastric carcinogenesis by oligonucleotide microarray.

METHODSU133A (Affymetrix, Santa Clara, CA) gene chip was used to detect the gene expression profile difference in T, P and C, respectively. Bioinformatics was used to analyze the detected results.

RESULTSWhen gastric cancer was compared with normal gastric mucosa, 766 genes were found,with a difference of more than four times in expression levels, including 530 up-regulated [Signal Log Ratio (SLR) > 2], and 236 down-regulated (SLR< -2). When P was compared with C, 64 genes were found, with a difference of more than four times in expression levels, including 50 up-regulated (SLR > 2), and 14 down-regulated (SLR< -2). Compared with C, a total of 143 genes with a difference of more than four times in expression levels both in T and P tissues. Of the 143 genes, 108 were up-regulated (SLR > 2), and 35 were down-regulated (SLR< -2).

CONCLUSIONSGene chip can reveal 143 same genes both in pericancerous mucosa and gastric mucosa. These genes may be related to the carcinogenesis and development of early gastric cancer.

Female ; Gastric Mucosa ; pathology ; Gene Expression Profiling ; methods ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; methods ; Precancerous Conditions ; Stomach Neoplasms ; genetics ; pathology

OBJECTIVETo prepare antibodies against pORF5 plasmid protein of Chlamydia trachomatis and develop double-antibody sandwich enzyme-linked immunosorbent assays (DAS-ELISAs) for the detection of genital C. trachomatis infections.

METHODSThe pORF5 protein was expressed in Escherichia coli and used to immunize BALB/c mice and New Zealand rabbits to produce monoclonal antibodies (mAbs) and polyclonal antibody (pAb) for DAS-ELISAs. Clinical samples from 186 urogenital infection patients (groups I) and 62 healthy donors (groups II) were detected in parallel by the DAS-ELISAs developed in this study and by IDEIA PCE commercial ELISA.

RESULTSTwo hybridoma cell lines, named 2H4 and 4E6, stably secreting specific mAbs against pORF5 were obtained. The mAb 2H4 was recognized by 32 (17.20%, positive recognition rate) and 25 (13.44%), mAb 2H4 by 0 (0%) and 2 (3.22%) samples from groups I and II, respectively. The sensitivities of mAbs 2H4 and 4E6 were 92.11% and 77.78% and the specificities were 100% and 96.88%, respectively in relation to the IDEIA PCE commercial ELISA. The sensitivities of detection for the DAS-ELISAs were 10 ng/mL (based on 2H4) and 18 ng/mL (based on 4E6).

CONCLUSIONTwo DAS-ELISAs were developed in this study that provided a feasible and effective assay that could be considered alternative tools for the serodiagnosis of C. trachomatis infection.

Adolescent ; Adult ; Chlamydia Infections ; diagnosis ; Chlamydia trachomatis ; pathogenicity ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Humans ; Male ; Middle Aged ; Urogenital System ; microbiology ; Young Adult

Adult ; Decompression, Surgical ; Diskectomy, Percutaneous ; Female ; Humans ; Intervertebral Disc Displacement ; surgery ; Laser Therapy ; Male ; Middle Aged

OBJECTIVETo screen the carcinogenesis associated genes in gastric carcinoma by gene chip.

METHODSU133A (Affymetrix Santa Clara, CA) gene chip was used to detect differentially expressed genes in tumor tissues, paratumor mucosa and normal mucosa. Bioinformatics was used to analyze the screened results.

RESULTSA total of 150 genes were detected with a difference of expression levels more than 3 times in paratumor mucosa compared with normal gastric mucosa, 130 of which were up-regulated and 20 down-regulated. According to the function classifications of the differentially expressed genes, the most common ones were enzyme and enzyme regulon activity associated genes(28, 18.7% ). The frequencies of nuclei acid binding activity associated genes,signal transduction associated genes and protein binding associated genes were 11.3%, 10%, and 8.7% respectively. Seventy-one differentially expressed genes were detected both in tumor tissues and paratumor mucosa compared with normal mucosa, 61 of which were up-regulated and 10 down-regulated. Among these 71 genes,e leven genes were localized on chromosome 19, 6 on chromosome 1, 2, 16, 17 respectively. No abnormal differentially expressed gene were detected on chromosome 5, 14, 22 and Y.

CONCLUSIONSThese 71 genes differentially expressed both in tumor tissues and paratumor mucosa may be associated with carcinogenesis of gastric carcinoma. The four kinds of genes associated with enzyme and enzyme regulon activity, nuclei acid binding activity, signal transduction, and protein binding should be the main genes for the study of carcinogenesis in gastric carcinoma.

Gastric Mucosa ; pathology ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Humans ; Oligonucleotide Array Sequence Analysis ; Stomach Neoplasms ; genetics ; pathology

OBJECTIVETo study the relationship of mesorectum with fasciae and nerves in the pelvic cavity and to specify the proper planes of dissection in total mesorectal excision.

METHODSTwenty-four pelvises (12 males and 12 females) harvested from cadavers were studied by dissection.

RESULTSThere were three planes surrounding the rectum as the visceral fascia, vesicohypogastric fascia and parietal fascia. The pelvic plexus and its branches situated between the visceral fascia and the vesicohypogastric fascia. Pelvic splanchnic nerves and hypogastric nerves were observed between the visceral fascia and the parietal fascia.

CONCLUSIONSThe posterior plane of total mesorectal excision lies between the visceral fascia and the parietal fascia. The lateral dissection should be conducted in a plane between the visceral fascia and the vesicohypogastric fascia. The proper planes for posterior and lateral resection can be identified by the hypogastric nerve and the pelvic plexus respectively.

Fascia ; anatomy & histology ; Fasciotomy ; Female ; Humans ; Male ; Mesentery ; anatomy & histology ; surgery ; Pelvis ; anatomy & histology ; surgery

BACKGROUNDFew studies have given suggestions on appropriate initiation insulin dosage when combined with oral antidiabetic drugs (OADs). This research was to investigate appropriate initiation insulin doses for insulin-naive type 2 diabetes patients with different combinations and the relationship between insulin dosage and relevant factors.

METHODSThis was a randomized, open-label, treat to target study. The target was 20% decrease of both fasting plasma glucose (FPG) and 2 hours post-breakfast blood glucose (P2hBG). One hundred and forty-seven insulin-naive Chinese patients recruited were randomly assigned to 3 groups: group A, patients received insulin monotherapy; group B, received insulin plus metformin (0.5 g, tid) and group C, received insulin plus metformin (0.5 g, tid) and pioglitazone (15 mg, qd). Insulin doses were initiated with a dose of 0.3 U×kg(-1)×d(-1) and titrated according to FPG and P2hBG till reached the targets.

RESULTSBoth the time of getting 20% reduction of FPG and P2hBG showed significant differences among the three groups. The time was shortest in Group C. The insulin doses needed to achieve glucose reduction of 20% in three treatment groups were (0.40 ± 0.04) U×kg(-1)×d(-1) for Group A, (0.37 ± 0.04) U×kg(-1)×d(-1) for Group B, and (0.35 ± 0.03) U×kg(-1)×d(-1) for Group C, respectively. Multiple linear stepwise regression analysis showed that insulin doses correlated with body weight, FPG, diabetes duration, age and history of sulfonylurea treatment. The standardized regression coefficients were 0.871, 0.322, 0.089, 0.067 and 0.063 (with all P < 0.05).

CONCLUSIONSTo achieve blood glucose's reduction of 20% within safety context, initial insulin doses were recommended as the following: 0.40 U×kg(-1)×d(-1) for insulin mono-therapy, 0.37 U×kg(-1)×d(-1) for insulin plus metformin treatment, and 0.35 U×kg(-1)×d(-1) for insulin plus metformin and pioglitazone treatment in Chinese type 2 diabetes outpatients. Body weight is found the most closely related factor to the insulin dosage.

Adult ; Aged ; Blood Glucose ; analysis ; Body Weight ; drug effects ; Diabetes Mellitus, Type 2 ; blood ; drug therapy ; Drug Therapy, Combination ; Female ; Humans ; Hypoglycemic Agents ; administration & dosage ; Insulin ; administration & dosage ; adverse effects ; therapeutic use ; Linear Models ; Male ; Metformin ; administration & dosage ; adverse effects ; Middle Aged ; Outpatients ; Regression Analysis ; Thiazolidinediones ; administration & dosage ; adverse effects

OBJECTIVETo clone the plasmid protein pORF8 of Chlamydia trachomatis and localize its expression in Chlamydia-infected cells.

METHODSpORF8 gene was amplified and cloned into pGEX-6p vector, and the pORF8 fusion protein was expressed in E.coli XL1 Blue. After purification with glutathione-conjugated agarose beads, the pORF8 fusion protein was used to immunize BALB/c mice to generate polyclonal antibodies against pORF8 protein. The antibodies obtained were used to localize the plasmid protein pORF8 in Chlamydia-infected cells with immunofluorescence assay (IFA).

RESULTSThe pORF8 gene 744 bp in length was successfully cloned and the GST fusion protein with a relative molecular mass of 54 000 was obtained. The cellular distribution pattern of the plasmid protein pORF8 was similar to that of the major outer membrane protein (MOMP), a known C. trachomatis inclusion body protein, but not to that of chlamydial protease-like activity factor (CPAF, a secreted protein).

CONCLUSIONThe plasmid protein pORF8 is localized on the bacterial organism as an inclusion body protein in C. trachomatis-infected cells. The cellular location of pORF8 protein can potentially provide important insights into the pathogenesis of C. trachomatis.

Animals ; Antibodies ; immunology ; Bacterial Proteins ; biosynthesis ; genetics ; Chlamydia Infections ; metabolism ; Chlamydia trachomatis ; chemistry ; genetics ; metabolism ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; HeLa Cells ; Humans ; Mice ; Mice, Inbred BALB C ; Plasmids ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology

OBJECTIVETo investigate the clinical characteristics of Segond fracture and its operative method and opportunity.

METHODSFrom June 2008 to December 2011, arthroscopic exploration was performed in 16 patients with Segond fracture. Six patients were explored at 1 week after injury ,their anterior cruciate ligament were broken completely combined with meniscus injury. Ten patients were explored at 8-10 weeks after injury, 4 patients with anterior and posterior cruciate ligament breakage completely, 4 patients with anterior cruciate ligament breakage completely,and 2 patients with anterior cruciate ligament breakage incompletely,among 10 cases, 8 cases of anterior cruciate ligament breakage completely combined with meniscus injury, 1 case of anterior cruciate ligament breakage incompletely without obviously meniscus injury, 1 case combined with fibular head fracture and lateral collateral ligament injury. All broken cruciate ligaments were rebuilt after arthroscopic exploration and meniscus injuries were sutured in 5 cases. Clinical effects were evaluated according to Lysholm-Gillquist scoring of knee joint function.

RESULTSSix patients were operated at 1 week after injury,their knee joint swelled obviously at 3 days after operation,unloaded blood oozing by joint puncture,and out-of-bed activity with assistance at the I week after operation. Other 10 patients were operated at 8-10 weeks after injury,no knee joint obviously swelled,no blood oozing was found by joint puncture ,and out-of-bed activity with assistance at 3 days after operation. All patients were followed up from 12 to 50 months with an average of 24 months. Postoperative Lysholm-Gillquist scoring of all patients were higher than preoperative and recovered well.

CONCLUSIONSegond fractures often associated with anterior cruciate ligament breakage and meniscus injury,it is important that early detection for treatment. The best time of cruciate ligament rebuilding and meniscus repairing may be at 8-10 weeks after injury.

Adult ; Anterior Cruciate Ligament ; surgery ; Anterior Cruciate Ligament Injuries ; Humans ; Male ; Menisci, Tibial ; surgery ; Middle Aged ; Tibial Fractures ; diagnosis ; surgery ; Tibial Meniscus Injuries

OBJECTIVETo evaluate the techniques and clinical applications of 16 multislice helical CT in colonic lesions.

METHODSEighty-one patients including 54 colorectal carcinomas, 5 adenomas, 1 non-Hodgkin's lymphoma, 6 inflammatory bowel diseases, and other 15 cases underwent volume scanning using 16 multislice helical CT. Four types of reconstruction included multiple planar reconstruction, shaded surface display, raysum, and CT virtual colonoscopy.

RESULTSComplete colon could be shown in all patients. The lesions' morphology, number, size, location, intestinal cavity, pericolonic changes, and other abdominal organs were satisfactorily shown by CT.

CONCLUSIONSSixteen multislice helical CT colonography is a valuable imaging technique for detecting colonic diseases. It is effective in diagnosis and treatment planning. It can display the portions of colon that is inaccessible at colonoscopy.

Adenocarcinoma ; diagnostic imaging ; Adenoma ; diagnostic imaging ; Adult ; Aged ; Aged, 80 and over ; Colonography, Computed Tomographic ; methods ; Colonoscopy ; Colorectal Neoplasms ; diagnostic imaging ; Female ; Humans ; Inflammatory Bowel Diseases ; diagnostic imaging ; Male ; Middle Aged ; Tomography, Spiral Computed ; methods