BACKGROUND: Primate is the best animal to establish a model of spinal implants. However, ethics and cost limit its application. Mixed-breed dogs have similar anatomic structures as human. Moreover, it is easy to obtain with low cost, so it may replace primate to serve as models.OBJECTIVE: To determine the feasibility of canine lumbar spine to establish the anterior interfixation model following measurements of related anatomic data.METHODS: A total of 9 adult healthy dogs were selected. The transversal diameter, sagittal diameter and height of vertebral body and disc were measured, and the artificial vertebral body replacement was performed. Bone graft fusion was observed. RESULTS AND CONCLUSION: The transversal diameter, sagittal diameter and height of vertebral body and disc increasedgradually from L_(1) to L_(7), and the height was greatly exceeded its sagittal diameter. All dogs survived, but one was paraplegia. Allthe others stood and acted in 12 h to 72 h after operation. The fusion effect was proved to be good by imageology and histology. The establishment of models was simple and cost-effective, and the biocompatibility of bone tissues and implants, as well as thebone tissue ingrowth can be observed. The lumbar spine of dog can be used as an anterior interfixation model in vitro test.

Objective To investigate the expression of Phosphorylated-signal transduction and activators of transcription 3 (P-STAT3) proteins in human prostate tissue from patients received re-peated biopsies. And consider the usefulness of detecting expression of P-STAT3 in early diagnosis of prostate cancer (PCA). Methods Fifty-eight patients (29 cases of PCA, and 29 cases of benign prostatic hyperplasia (BPH)) who had received repeated biopsies were involved in this study. Immu-nolabelling has been carried out on PCa patients' samples of cores from initially negative biopsies, typ-ical cores from cancer field, and other cores of the same batch biopsies showing no sign of prostate cancer. BPH patients' samples of cores from initial biopsies were set as control. All specimens were done immunohistochemistry stain with anti-P-STAT3 monoclonai antibody. The association of P-STAT3 expression in prostate tissues with the pathology result was evaluated. Results Compared with 10.3% in specimens of patients free of prostate cancer, the positive rate of anti-P-STAT3 stained in typical cores from cancer field, other cores of the same batch biopsies showing no sign of prostate cancer, and cores from initially negative biopsies, was 93.1 % (27/29), 82.8 % (24/29) and 86.2 % (25/29), respectively. There were significant differences of these values between former and laters' (X2=60.123,P=0.000). If P-STAT3 positive in tissue of initially biopsies was considered as the di-agnostic standard of prostate cancer, then it would show a relatively high sensitivity (86.2%) and specificity (89.7%). Conclusion IHC stain for P-STAT3 in prostate biopsy samples could be served as an adjunct to the current diagnostic approach to prostate biopsy for early diagnosis of pros-tate cancer.

ObjectiveTo optimize human Hexastatin gene,to express,purify protein and conduct activity experimental research,and to provide a theoretical basis for further study of Hexastatin.MethodsHuman Hexastatin gene was optimized and synthesized.It was connected to the pET28a expression vector,induced to express by isopropyl β-D-1-thiogalactopyranoside(IPTG),and optimized induction conditions.After the ultrasonication of bacterial cells and inclusion bodies,the recombinant fusion protein was purified with Ni-NTA chromatographic column,analyzed and identified by SDS-PAGE and Western Blot,and conduct activity experimental research in vitro by MTT.ResultsConstructed production was pET28a-Hexastatin expression plasmid.The human Hexastatin protein was expressed in E.coli BL21 the high level and accounted for 45.1％ of the total bacterial protein.The purification of recombinant protein purified with Ni-NTA chromatographic column reached 90％,and the concentration was 80 μg/ml.Human Hexastatin protein can restrain the growth of C6,MCF-7 and human vascular endothelial cell (HMEC) cells,and inhibition ratio reach to 72.9％±3.6％,48.8％±2.9％,52.7％±2.5％,respectively through MTT test.ConclusionThe optimized human Hexastatin protein was expressed successfully,which confirmed the inhibition to tumour cells.It is a new way for anti-angiogenesis therapy of tumour.

Objective To study the clinicopathologic characteristics of triple-negative breast cancer (TNBC) and its value in the prediction of prognosis. Method In this study,500 cases of female breast cancers were examined immunohistochcmically for the TNBC. The clinicopathologic characteristics of the 243 TNBC cases were inspected. Results TNBC accounted for 17.6% (88/500) of the 500 breast cancers. The histological types of the TNBC included mainly infihrative ductal carcinoma, metaplastic carcinoma and medullar carcinoma. Among those, histological grade Ⅲ accounted for 72.7% (64/88) of all the TNBC and was more common than that in hormone receptor positive breast cancers (HR+ group ) and Her-2 overexpression breast cancers (Her-2 group)(P=0.000). The positive rates of CK5/6 and EGFR in the TNBC were 30.7% (27/88) and 34.1% (30/88), respectively. The positive rates of ERCC1 and KIT in the TNBC were 28.4% (25/88) and 34.1% (30/88), respectively, Both of which were higher than those in the HR + group and Her-2 group, respectively (P=0.032 and P=0.026). 3-year survival rate of the TNBC was 71.5% and it was lower than that of HR group (P=0.021) and not significantly different from that of Her-2 group (P=0.474). Conclusions TNBC is the breast cancer with high aggressive pathologic futures and poor prognosis. EGFR and ERCC1 expression were positive in a portion of TNBC cases.

OBJECTIVE: To determine the role of fragile histidine triad (FHIT) and MDM2 in carcinogenesis of oral submucous fibrosis (OSF). METHODS: The expression of FHIT and MDM2 was examined by immunohistochemical S-P method in 44 OSF cases, 15 canceration tissues of OSF, and 10 normal oral mucosa tissues. RESULTS: The expression of FHIT was positive in the normal oral mucosa epithelium. The positive expression of FHIT decreased in the OSF and canceration tissues of the OSF.The rate of FHIT positive expression was significantly lower in canceration tissues of OSF than that of the OSF (P < 0.05). The expression of MDM2 was negative in normal oral mucosa epithelium. The positive expression of MDM2 increased in the OSF and canceration tissues of the OSF, and the rate of MDM2 positive expression was significantly higher in the canceration tissues of OSF than that of the OSF (P < 0.05). CONCLUSION The loss of FHIT and over-expression of MDM2 may play an important role in the carcinogenesis of OSF.
Acid Anhydride Hydrolases ; genetics ; metabolism ; Female ; Humans ; Immunohistochemistry ; Male ; Mouth Mucosa ; metabolism ; Mouth Neoplasms ; metabolism ; Neoplasm Proteins ; genetics ; metabolism ; Oral Submucous Fibrosis ; metabolism ; Proto-Oncogene Proteins c-mdm2 ; genetics ; metabolism

OBJECTIVETo analyze the pharmacokinetics of intraperitoneal chemotherapy with mitomycin C (MMC) bound to activated carbon particles.

METHODSA nude mouse model with transplanted human gastric cancer was established. The mice were given MMC by i.v. or intraperitoneal (i.p.) injections, or given i.p. MMC bound to activated carbon particles (MMC-CH). Pharmacokinetic assays were carried out at different time points (0.5, 1, 2, 3, 6, 12 and 24 h) in 7 mice per each time point, to compare the MMC concentrations revealed by the above mentioned methods.

RESULTSThe MMC concentration in peritoneal exudate, omentum and lymph nodes of MMC-CH group was significantly higher than that of MMC solution i.p. group and MMC i.v. group (P < 0.001). On the other hand, the MMC level in serum was significantly lower than that in two control groups (P < 0.001). High MMC level was maintained longer than 24 hours in the MMC-CH group. Intraperitoneal chemotherapy with MMC solution resulted in a low MMC concentration in serum, peritoneal exudates and lymph nodes, and only a transient high level of MMC in the omentum. After i.v. administration, a significantly higher level of MMC concentration occurred in the serum, but only a shortly increased concentration of MMC in the omentum, and lower concentration in peritoneal exudate and lymph nodes as compared with those in the other two groups (P < 0.001).

CONCLUSIONHigh concentration of MMC in peritoneal exudate, omentum and lymph nodes maintained longer than 24 hours and a significantly lower MMC serum concentration can be achieved by administration of intraperitoneal administration of MMC bound to activated carbon particles.

Animals ; Antibiotics, Antineoplastic ; administration & dosage ; pharmacokinetics ; Charcoal ; administration & dosage ; pharmacokinetics ; Female ; Humans ; Injections, Intraperitoneal ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Mitomycin ; administration & dosage ; pharmacokinetics ; Neoplasm Transplantation ; Stomach Neoplasms ; drug therapy ; metabolism ; pathology

OBJECTIVE: To investigate the gut microbiota in newly diagnosed IgA nephropathy patients with chronic kidney disease (CKD) stages 1-2 and the association between the gut microbiota and the clinical risk factors of IgA nephropathy. METHODS: Fresh fecal samples were collected from nineteen newly diagnosed IgA nephropathy patients with CKD stages 1-2 and fifteen age- and sex-matched healthy controls. Fecal bacterial DNA was extracted and microbiota composition were characterized using 16S ribosomal RNA (16S rRNA) high-throughput sequencing for the V3-V4 region. The Illumina Miseq platform was used to analyze the results of 16S rRNA high-throughput sequencing of fecal flora. At the same time, the clinical risk factors of IgA nephropathy patients were collected to investigate the association between the gut microbiota and the clinical risk factors. RESULTS: (1) At the phylum level, the abundance of Bacteroidetes was significantly reduced (P=0.046), and the abundance of Actinobacteria was significantly increased (P=0.001). At the genus level, the abundance of Escherichia-Shigella, Bifidobacte-rium, Dorea and others were significantly increased (P < 0.05). The abundance of Lachnospira, Coprococcus_2 and Sutterella was significantly reduced (P < 0.05). (2) There was no significant difference in the abundance of gut microbiota between the newly diagnosed IgA nephropathy patients and the healthy control group (P>0.05), but there were differences in the structure of the gut microbiota between the two groups. The results of LEfSe analysis showed that there were 16 differential bacteria in the newly diagnosed IgA nephropathy patients and healthy controls. Among them, the abundance of the newly diagnosed IgA nephropathy patients was increased in Enterobacteriales, Actinobacteria, Escherichia-Shigella, etc. The healthy control group was increased in Bacteroidetes and Lachnospira. (3) The result of redundancy analysis (RDA) showed that Bifidobacterium was positively correlated with serum IgA levels, 24-hour urinary protein levels and the presence of hypertension. Lachnoclostridium was positively correlated with the presence of hypertension. Escherichia-Shigella was positively correlated with urine red blood cells account. Bifidobacterium was positively correlated with the proliferation of capillaries. Faecalibacterium was positively correlated with cell/fibrocytic crescents. Ruminococcus_2 was positively correlated with mesangial cell proliferation, glomerular segmental sclerosis and renal tubular atrophy/interstitial fibrosis. CONCLUSION The gut microbiota in the newly diagnosed IgA nephropathy patients with CKD stages 1-2 is different from that of the healthy controls. Most importantly, some gut bacteria are related to the clinical risk factors of IgA nephropathy. Further research is needed to understand the potential role of these bacteria in IgA nephropathy.
Humans ; Gastrointestinal Microbiome ; RNA, Ribosomal, 16S/genetics* ; Glomerulonephritis, IGA ; Bacteria/genetics* ; Risk Factors ; Renal Insufficiency, Chronic

OBJECTIVETo prepare and characterize monoclonal antibodies (mAb) and polyclonal antibodies against nucleocapsid (N) protein of severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV) and to establish antibodies-based sandwich ELISA for detecting N protein of SARS-CoV, which might apply to early diagnosis of patients with SARS-CoV infection.

METHODSBALB/c mice were immunized with purified recombinant N protein of SARS-CoV for producing mAbs, and New Zealand white rabbits were immunized for producing polyclonal antibodies. The identification of antibodies was performed using indirect enzyme-linked immunosorbent assay (ELISA), indirect fluorescent-antibody assay (IFA), and Western immunoblotting. Capturing and detecting antibodies were selected by pairing the mAbs and polyclonal antibodies one by one and an antibodies-based sandwich antigen capture ELISA was used for detecting N antigen of SARS-CoV.

RESULTSNine mAbs and hyperimmune rabbit polyclonal antibodies, specifically against SARS-CoV nucleocapsid protein were obtained. Using paired ELISA assay, three mAbs N1E8, N8E1 and N10E4 were selected as capturing antibody and rabbit polyclonal antibodies as detecting antibody then triple antibodies-based sandwich ELISA was established following horseradish peroxidase (HRP)-conjugated goat anti-rabbit immunoglobulin G. The recombinant N protein was used as a standard to establish a detection sensitivity of approximated 50 pg/ml with this assay. When tested with 420 serum specimens from serologically confirmed SARS patients, the positive rates of serum N protein were 90.1%, 23% and 0%, in which sera collected from 1 to 10 days, 11 to 20 days and beyond 21 days respectively after the onset of symptoms. The specificity of the assay was 99.86% in 715 control serum specimens. There was no cross-reaction with other respiratory viruses and coronaviruses.

CONCLUSIONSpecific and high affinity mAbs and rabbit polyclonal antibodies were obtained. By paired and optimized sandwich ELISA, a sensitive and specific antigen capture ELISA was established for detecting N antigen of SARS-CoV, which might apply to early diagnosis, source tracing and epidemiological studies of SARS.

Animals ; Antibodies, Monoclonal ; biosynthesis ; Antibodies, Viral ; blood ; Enzyme-Linked Immunosorbent Assay ; Humans ; Mice ; Mice, Inbred BALB C ; Nucleocapsid ; immunology ; Rabbits ; SARS Virus ; immunology ; isolation & purification ; Sensitivity and Specificity ; Severe Acute Respiratory Syndrome ; virology

PURPOSE: To investigate and compare the effects of propofol and midazolam on inflammation and oxidase stress in children with congenital heart disease undergoing cardiac surgery. MATERIALS AND METHODS: Thirty-two ASA class I-II children with congenital heart disease undergoing cardiac surgery were randomly divided into two groups: propofol combined with low dose fentanyl (PF group, n = 16) and midazolam combined with low dose fentanyl (MF group, n = 16). Tracheal extubation time and length of Intensive Care Unit (ICU) stay were recorded. Blood samples were taken before operation (T0), at 2 h after release of the aorta cross-clamp (T3) and at 24 h after operation (T4) to measure interleukin 6 (IL-6), IL-8, superoxide dismutase (SOD) and malondialdehyde (MDA) levels. Myocardium samples were collected at 10-20 min after aorta cross-clamp (T1) and at 10-20 min after the release of the aorta cross-clamp (T2) to detect heme oxygenase-1 (HO-1) expression. RESULTS: Tracheal extubation time and length of ICU stay in PF group were significantly shorter than those of the MF group (p < 0.05, respectively). After cardiopulmonary bypass, IL-6, IL-8 and MDA levels were significantly increased, and the SOD level was significantly reduced in both two groups, but PF group exhibited lower IL-6, IL-8 and MDA levels and higher SOD levels than the MF group (p < 0.05, respectively). The HO-1 expression in the PF group was significantly higher than that in MF group at the corresponding time points (p < 0.05, respectively). CONCLUSION: Propofol is superior to midazolam in reducing inflammation and oxidase stress and in improving post-operation recovery in children with congenital heart disease undergoing cardiac surgery.
Anesthesia, Intravenous/*adverse effects ; Anesthetics, Intravenous/*adverse effects ; Cardiac Surgical Procedures/*adverse effects ; Child ; Female ; Heart Defects, Congenital/*surgery ; Heme Oxygenase-1/blood ; Humans ; Inflammation/*chemically induced ; Interleukin-6/blood ; Interleukin-8/blood ; Male ; Malondialdehyde/blood ; Midazolam/*adverse effects ; Oxidative Stress/*drug effects ; Propofol/*adverse effects ; Superoxide Dismutase/blood

OBJECTIVETo investigate the effect of prescription compatibility on the pharmacokinetics of components from Dachengqi Decoction (DCQD, ) in rats.

METHODSTwenty-four male rats were randomly and equally divided into the DCQD group, Dahuang (Radix et Rhizoma Rhei, Polygonaceae) group, Houpo (Magnolia officinalis Rehd., Magnoliaceae) group, and Zhishi (Fructus Aurantii Immaturus, Rutaceae) group. The blood samples were collected before dosing and subsequently at 10, 15, 20, 30, 45 min, 1, 2, 4, 8, and 12 h following gavage. The levels of aloe-emodin, rhein, emodin, chrysophanol, honokiol, magnolol, hesperidin, and naringin in rat serum were quantified using a liquid chromatography tandem mass spectrometry (LC-MS/MS) method for pharmacokinetic study.

RESULTSThe area under the curve (AUC), mean retention time (MRT), the peak concentration (C(max)) of aloe-emodin, rhein, emodin, and chrysophanol in the DCQD group were significantly different compared with the Dahuang group (P <0.05, respectively). The mean plasma concentration, C(max), and the absorption of Dahuang's component in the DCQD group were obviously lower at each time point than those in the Dahuang group, while the elimination process of Dahuang's component was obviously delayed (P <0.05). Half-lives of aloe-emodin, chrysophanol, and rhein were also extended in the DCQD group (P <0.05, respectively). In the DCQD group, the mean plasma concentration, AUC, C(max) and absorption of honokiol, and magnolol were significantly lower (P <0.01, respectively) at each time point than those in the Houpo group, while the drug distribution half-life time (T(1/2α)), the drug eliminated half-life time (T(1/2β)), MRT, and time of peak concentration (T(max)) were significantly delayed (P <0.05, respectively). Pharmacokinetic parameters of hesperidin and naringin in the Zhishi group were not significantly different as compared with the DCQD group (P >0.05, respectively), while the MRT of naringin was significantly longer.

CONCLUSIONSThe compatibility in Chinese medicine could affect the drug's pharmacokinetics in DCQD, which proves that the prescription compatibility principle of Chinese medicine formulations has its own pharmacokinetic basis.

Administration, Oral ; Animals ; Anthraquinones ; administration & dosage ; blood ; pharmacokinetics ; Biphenyl Compounds ; administration & dosage ; blood ; pharmacokinetics ; Drug Incompatibility ; Emodin ; administration & dosage ; blood ; pharmacokinetics ; Flavanones ; administration & dosage ; blood ; pharmacokinetics ; Hesperidin ; administration & dosage ; blood ; pharmacokinetics ; Lignans ; administration & dosage ; blood ; pharmacokinetics ; Male ; Plant Extracts ; administration & dosage ; blood ; chemistry ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley