Biopsy ; Histological Techniques ; instrumentation ; methods ; Humans ; Indicators and Reagents ; Paraffin Embedding ; Ultrasonics

Objective:To generate sex hormone binding globulin(SHBG) conditional knockout mice model.In order to investigate the physiological function of SHBG in vivo and to provide experimental means for the study of the relationship between SHBG and gestational diabetes mellitus.Methods:The mouse genomic DNA sequence of SHBG was verified through bioinformatic analysis.According to the SHBG genomic DNA sequence,the gene targeting and knockout vector were constructed.Transfection of the vectors to ES cells by electroporation was performed according to common protocol.Positive ES cells were screened and identified by PCR.Therefore,the dual selected ES cells were microinjected into blastula,then blastula transplantations into the host mice.The chimeric mice were mated with C57BL/6J mice,and the Flox mice were obtained after screening.The Flox mice were hybridized with EIIA-Cre transgenic mice,and the progeny of the SHBG gene knockout (SHBG-/-) mice were obtained by autocopuation for several times.Results:Several Flox homozygous mice and SHBG gene knockout mice were successfully obtained.Compared with control mice,homozygous mice of SHBG gene knockout were well developed and had reproductive ability.The growth and development of SHBG knockout mice were not significantly different from that of wild type mice.Conclusion:Homozygous mice model of SHBG gene knockout was successfully established,which laid the foundation for further study of the role of SHBG in the gestational diabetes.The SHBG gene knockout mouse model was successfully established and the preliminary phenotypic analysis was performed,which laid the foundation for further study on the role of SHBG in gestational diabetes mellitus.SHBG gene knockout mice were normal in appearance.Due to the limited number of samples and many unknown biological characteristics of gene knockout mice,it needs further study.

Bufalin is an active compound of the traditional Chinese medicine Chansu, which exhibits significant anti-tumor activities in many solid tumors and leukemia cell lines. Bufalin could introduce apoptosis, reverse drug-resistance, and prevent migration and invasion of tumor cells. This paper reviewed the latest research progress of the in vitro and in vivo anti-tumor effect and mechanism of bufalin on a series of cancers, such as hepatocellular carcinoma, lung cancer, colon cancer, gastric cancer, leukemia, bladder cancer, and its formulation study is also summarized for the reference of its further study and application.
Animals ; Antineoplastic Agents ; chemistry ; pharmacology ; therapeutic use ; Bufanolides ; chemistry ; pharmacology ; therapeutic use ; Chemistry, Pharmaceutical ; methods ; Neoplasms ; drug therapy ; pathology

Abstract: Objective To express and purify MPT83 protein of Mycobacterium tuberculosis and evaluate its application value in immunological diagnosis of tuberculosis (TB) using clinical samples. Methods Using Mycobacterium tuberculosis (Mtb) H37Rv genome as the template, Mtb mpt83 gene was amplified by PCR and connected to PET-21a (+) to construct prokaryotic expression vector, and then transferred into E.coli DH5α. The positive colonies were picked out and retained. The recombinant plasmid pET-mpt83 of the strain with positive colony PCR was extracted, identified by double digestion, and the samples of the positive colonies were sent for sequencing. The correctly sequenced plasmids then were transferred into BL21 competent cells for induction, expression and purification with nickel column affinity chromatography. The purified products were identified by 12 alkyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. Mouse polyclonal antiserum was prepared by immunizing mice with purified protein. 8 patients clinically diagnosed as tuberculosis pleural effusion (TB group) and 8 adenocarcinomas patients (CA group) were enrolled and their pleural effusion and plasma were collected. 8 healthy people (HC group) were enrolled as the control group and their plasma were collected. An indirect ELISA was used to detect the level of specific antibodies recognizing MPT83 protein in the samples. Results Mtb MPT83 protein was successfully expressed and purified. The serum titer of MPT83 mouse polyclonal antibody was as high as 1∶1 280 000. The plasma levels of MPT83 antigen specific antibodies in TB group were significantly higher than those in HC group (P<0.05), while the plasma levels of MPT83 antigen specific antibodies in CA group were not significantly different from those in HC group (P>0.05). Compared with the HC group, there was no significant difference in pleural fluid in both the TB and CA groups (P>0.05). The ROC curve was used to analyze the OD values of plasma in TB group and HC group, and the area under the curve was greater than 0.7, showing high diagnostic efficacy. Conclusion　MPT83 protein has high antigen specificity and immunogenicity, which has great application value in the immunological diagnosis of tuberculosis.

OBJECTIVEThe aim of the study was to establish a cephalometric normal standard value for Korean in Yanbian China by means of Steiner analysis.

METHODS72 cases of Korean adults in Yanbian with normal occlusion were selected, standard lateral cranial X-ray films were taken and cephalometric Steiner analysis was performed. The data was compared with Korean in Soul of Republic of Korea with normal occlusion.

RESULTS1) Cephalometric Steiner normal standard values, standard deviation and Cheveron analysis chart were established. 2) 1-NB angle of male was significantly bigger than that of female, while GoGn-SN angle of female was significantly bigger than that of male (P<0.05), respectively. 3) There were significant differences on SNA, SNB, ANB, SND, [symbol: see text]-NA (mm),1-NB (except woman) compared with same age group Korean presented by In-Chool Park (P<0.05).

CONCLUSIONThere is a significantly different between male and female Koreans with normal occlusions in Yanbian China. This experiment filled in the blank of stomatology research in Yanbian Korean autonomous region. In the mean while, it completed and supplemented the normal occlude data of different nations in China. The present paper also provided the reference for clinic and scientific research on orthognathic surgeon. At the same time, the experiment provided the evidence for the further research.

Adult ; Cephalometry ; China ; Dental Occlusion ; Female ; Humans ; Male ; Reference Values ; Skull

OBJECTIVETo evaluate the efficacy of abdominal acupuncture for adhesion-stage shoulder periarthritis.

METHODSOne hundred and fifty-seven cases of shoulder periarthritis were randomly divided into an abdominal acupuncture group (79 cases) and a body acupuncture group (78 cases). The abdominal acupuncture was applied at Zhongwan (CV 12), Shangqu (KI 17) and Huaroumen (ST 24) in the abdominal acupuncture group while conventional acupuncture was applied at Jianyu (LI 15), Jianliao (TE 14) and Jianzhen (SI 9) in the body acupuncture group. The treatment was given three times a week for both groups and ten times made an observation course. Before and after treatment, visual analogue scale (VAS) was adopted for pain assessment and functional activity score (Mallet score) was used for shoulder joint function assessment for all the patients. Also the efficacy of both groups was compared.

RESULTSThe total effective rate in the abdominal acupuncture group was 92.4% (73/79), which was superior to 71.8% (56/78) in the body acupuncture group. The score of VAS after the treatment was 2.58 +/- 1.64 in the abdominal acupuncture group while 3.12 +/- 1.93 in the body acupuncture group, which had no statistical significance between each other (P > 0.05). The functional activity score after the treatment was 8.34 +/- 3.02 in the abdominal acupuncture group while 7.49 +/- 3.36 in the body acupuncture group, which had no statistical significance between each other (P > 0.05).

CONCLUSIONThe abdominal acupuncture is an ideal treatment for adhesion-stage shoulder periarthritis, which has better total efficacy than conventional acupuncture. It achieves the same effect in relieving pain and improving functional activity as conventional acupuncture, but also has an advantage at fast selection of acupoint and less discomfort of needling sensation.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Periarthritis ; physiopathology ; therapy ; Shoulder ; physiopathology

Objective　To investigate the relationship between the pathological changes and the early anastomosis of the blood vessels in blast-wounded maxillofacial region. Methods　A total of 20 dogs were regionally exploded in the left maxillofacial region by KTY-04 electric detonator. Specimens of blood vessels were removed at different distance from the edge of the wounded area at both right after (group A) and 72 h (group B ) after wounded and observed. Free skin flaps with saphenous artery were transplanted to the wounded areas. One week, 2 weeks and 3 months after anastomosis, specimens were taken from the site of anastomosis and observed. Results　Falling off of most vascular endothelial cells and the disappearance of most internal elastic membrane of the artery were observed 3 cm from the wounded area in group A with 5 free flaps died. However in group B almost normal condition was seen at the same region. 12 from 15 flaps survived successfully. Conclusion　The blood vessels begin to proliferate and repair 3 d after blast wound and this may provide the pathological basis for the possible success in the transplantation of the free flaps with blood vessels.

OBJECTIVE: To investigate the genetic polymorphism in the 5' and 3' region of TPH2 gene of Northern Chinese Han population and to explore its application value in forensic medicine. METHODS: The sequence variants and the genetic polymorphisms of 6 SNP loci (rs4570625, rs11178997, rs11178998, rs41317118, rs17110747 and rs41317114) within a 905 bp 5' flanking region and a 1,104bp 3' flanking region of TPH2 gene were analyzed by DNA sequencing in a total of 244 unrelated healthy individuals in Northern Chinese Han population. The statistical analysis was carried out by Haploview v4.2 software. RESULTS: The genotypic distributions of the 6 SNP loci in the TPH2 gene were in accordance with Hardy-Weinberg equilibrium. One C/T variant in 92922 site was found. There was a high linkage disequilibrium among the 3 SNP loci (rs4570625, rs11178997 and rs11178998) in the 5' region and the 3 SNP loci (rs41317118, rs17110747 and rs41317114) in the 3' region of TPH2 gene, respectively. The parameters of population genetics of 6 SNP loci were obtained. CONCLUSION There are great polymorphisms in the 5' and 3' region of TPH2 gene in Northern Chinese Han population, which could be used as genetic indexes for association analysis of the related diseases, as well as for forensic individual identification and paternity testing.
3' Untranslated Regions ; 5' Untranslated Regions ; Asian People/genetics* ; China/ethnology* ; Forensic Genetics ; Gene Frequency ; Genetics, Population ; Genotype ; Humans ; Linkage Disequilibrium/genetics* ; Microsatellite Repeats ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Tryptophan Hydroxylase/genetics*

OBJECTIVETo evaluate the application of multi-detector row CT (MDCT) and CT angiography (CTA) for detecting early signs of acute bowel ischemia (ABI) in experimental porcine models.

METHODSTwelve pigs were assigned to four groups with 3 in each group. The digital subtraction angiography of superior mesenteric artery (SMA) and the embolization of branches of SMA with gelatin sponge and blood clot were performed by percutaneous transfemoral artery puncture and catheterization. MDCT pre- and post-contrast scanning in the arterial, venous and delay phase and CTA with three-dimensional reconstruction were carried out at pre-operation, 3 h, 6 h, 9 h, and 12 h after occlusion. The normal mesenteric vascular anatomy, arterial occlusion, mesentery and bowel changes, and dynamic change were evaluated.

RESULTSABI changes were identified pathologically in all the 12 experimental pigs, and the severity of ischemia increased over time after embolization. CTA showed all 57 embolized branches of SMA and 29 of 34 unoccluded arterial branches with 5 false-positive vessel occlusions. The sensitivity and specificity of CTA were 100% and 85.3%, respectively. Thin-slab maximum intensity projection (TSMIP) revealed the disappearance of distal comb-like vessel branches and brush-like vasa recta, which were clearly delineated in the normal bowel segments. Using this criterion, TSMIP correctly defined 23 of 24 ischemic bowel segments and all the 12 normal bowel segments with a sensitivity of 95.8% and a specificity of 100%.

CONCLUSIONSMDCT and CTA reliably define normal and occluded mesenteric vessels in the pig. It can easily detect ischemic bowel segment by identified early changes of ischemia. The early direct ischemic signs are occluded vessels, the disappearance of distal comb-like branches or brush-like vasa recta, and poor bowel enhancement. The early indirect sign is bowel dilatation with fluid collection.

Angiography ; methods ; Animals ; Female ; Intestinal Diseases ; diagnostic imaging ; etiology ; Ischemia ; diagnostic imaging ; etiology ; Mesenteric Arteries ; diagnostic imaging ; Mesenteric Vascular Occlusion ; complications ; diagnostic imaging ; Mesentery ; blood supply ; Swine ; Tomography, X-Ray Computed ; methods

OBJECTIVEMajor histocompatibility complex class I C-related molecules A and B (MICA and MICB) are innate immune system ligands for the NKG2D receptor expressed by natural killer cells and activated CD8(+)T cells. Our previous study showed that 5-aza-2'-deoxycytidine (5-aza-dC), a DNA methyltransferase inhibitor, can induce the expression of MICB and sensitized cells to NKL-cell-mediated cytolysis. The aim of this study was to determine the expression level of MICA in HepG2 cells (an HCC cell line) and L02 cells ( a normal liver cell), and to investigate the effect of 5-aza-dC on MICA expression in HepG2 cells.

METHODSCells were treated with 5-aza-dC, caffeine and ATM-specific siRNA. The cell surface MICA protein on HepG2 cells and L02 cells was determined using flow cytometry. The mRNA level was detected using real time RT-PCR.

RESULTSMICA was undetectable on the surface of L02 cells, but was highly expressed on HepG2 cells. MICA expression was upregulated in response to 5-aza-dC treatment (P less than 0.05), and the upregulation of MICA was partially prevented by pharmacological or genetic inhibition of ataxia telangiectasia mutated (ATM) kinase (P less than 0.05).

CONCLUSIONOur data suggest that 5-aza-dC induces the expression of MICA by a DNA damage-dependent mechanism.

Ataxia Telangiectasia Mutated Proteins ; Azacitidine ; analogs & derivatives ; pharmacology ; Caffeine ; pharmacology ; Carcinoma, Hepatocellular ; metabolism ; Cell Cycle Proteins ; antagonists & inhibitors ; metabolism ; Cell Line ; Cell Membrane ; metabolism ; DNA Damage ; DNA-Binding Proteins ; antagonists & inhibitors ; metabolism ; Flow Cytometry ; Hep G2 Cells ; Hepatocytes ; metabolism ; Histocompatibility Antigens Class I ; genetics ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; Protein-Serine-Threonine Kinases ; antagonists & inhibitors ; metabolism ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Suppressor Proteins ; antagonists & inhibitors ; metabolism ; Up-Regulation