OBJECTIVETo test the different contrctile responses of extracellular nucleotides, such as ATP, UTP and nucleotide uridine adenosine tetraphosphate (Up4A) in gastric longitudinal muscle (LM) and circular muscle (CM). Examined the effect of P2X and P2Y receptor antagonists (in this study, we used IP5I and suramin) and cyclooxygenase inhibitor (indomethacin) on Up4A induced contractile responses in LM and CM.

METHODSThe rats were sacrificed and the stomachs were opened to gain LM and CM. Using organ bath system to assess contrctile responses of smooth muscle.

RESULTSUp4A could induce contractile responses in both CM and LM, which were similar with ATP and UTP. IP5 did not attenuate Up4A could induce contractions in both LM and CM, but suramin and indomethacin significantly inhibited Up4A contraction in CM, but not in LM.

CONCLUSIONOur results suggest that extracellular nucleosides and their inhibitors induce different responses between LM and CM.

Adenosine Triphosphate ; pharmacology ; Animals ; Dinucleoside Phosphates ; pharmacology ; Indomethacin ; Muscle Contraction ; Muscle, Smooth ; physiology ; Nucleotides ; pharmacology ; Rats ; Suramin ; Uridine Triphosphate ; pharmacology

Abstract: Objective To explore the threshold of ALT for initiating antiviral therapy in HBV infected patients, and to provide a basis for initiating antiviral therapy in chronic HBV-infected patients. Methods This retrospective cohort study recruited 707 consecutive treatment-naïve chronic hepatitis B (CHB) patients undergoing diagnostic liver biopsy in the department of infectious diseases of the Affiliated Hospital of Yan′an University from October 2013 to August 2018. Liver biopsy specimens were obtained under ultrasound guidance using Menghini 16G disposable needles. The METAVIR scoring system, which is commonly used internationally, was used to divide the patients into the group with mild liver tissue injury and the group with significant liver tissue injury, and the alanine aminotransferase (ALT) levels were measured separately. Receiver operating characteristic (ROC) curve and Mann-Whitney U test were used to evaluate the diagnostic value of ALT for significant liver tissue injury under different demographic characteristics. Results Of 707 patients, 292 (41.30%) had significant liver tissue injury confirmed by liver biopsy (METAVIR ≥A2 and/or F2). When the ULN of ALT was set to NICE criteria (30 U/L for males, 19 U/L for females), AASLD criteria (35 U/L for males, 25 U/L for females) and EASL or APASL criteria (40 U/L for males and females), CHB patients with 30 years olds, 22 U/L for HBeAg negative and 31 U/L for HBeAg positive patients. Conclusions The threshold of ALT for initiating antiviral therapy in chronic HBV patients should be individualized, especially should be down-regulated for the females, olders and HBeAg-negative patients.

OBJECTIVETo observe the clinical efficacy of methotrexate (MTX) combined with total glucosides of Peony (TGP) on rheumatoid arthritis (RA).

METHODSSixty-one RA patients were divided into 2 groups, 30 patients in the MTX group were only administered orally with MTX, while 31 patients in the combined treated group were treated with MTX plus TGP, the therapeutic course for both groups was 3 months.

RESULTSThe total effective rate was 90%, 94%, 100% in the MTX plus TGP group, and 87%, 90%, 94% in the MTX group at 4, 8 and 12 weeks after treatment respectively, comparison of the therapeutic effect between the two groups showed insignificant difference (P > 0.05). The erythrocyte sedimentation rate (ESR) and the level of C-reactive protein were significantly decreased in both groups, but the decrement in the MTX plus TGP group was more than those in the MTX group.

CONCLUSIONMTX plus TGP treatment is characterized by quick initiating, with stable clinical efficacy, few side effects and high compliance, it is more suitable for aged RA patients.

Adult ; Aged ; Anti-Inflammatory Agents, Non-Steroidal ; therapeutic use ; Arthritis, Rheumatoid ; drug therapy ; Drug Therapy, Combination ; Female ; Glucosides ; therapeutic use ; Humans ; Immunosuppressive Agents ; therapeutic use ; Male ; Methotrexate ; therapeutic use ; Middle Aged ; Paeonia ; chemistry ; Phytotherapy

OBJECTIVETo investigate various methods for constructing soybean lecithin (SL)-based vesicles and evaluate the permeation-enhancing effect of SL-based vesicles on the penetration of insulin through buccal mucosa.

METHODSThe ultrasonic method, high speed shear method and high pressure homogenization method were respectively used to prepare the SL-based vesicles, and the particle size of the vesicles was measured with photon correlation spectrometry (PCS). The penetration rate of insulin through porcine buccal mucosa was investigated with the Valia-Chien diffusion cells.

RESULTSThe average particle sizes of 3 formulations of SL-based vesicles were 97.39, 85.60, and 100.60 nm when prepared by ultrasonic method, and were 58.7, 88.7, and 91.9 nm when prepared by high pressure homogenization method. Both vesicles presented good stability. However, the SL-based vesicles prepared by high speed shear method had larger average diameters and were found to be unstable. Transmission electron microscopy showed that SL-based vesicles had a spherical shape and the result accorded with PCS. The permeation flux of insulin of formulation 1 and control solution were 0.0024 and 0.0008 IU x ml(-1) x min(-1), respectively. The accumulative amount of formulation 1 at 180 min was (0.436 +/- 0.010 ) IU x ml(-1), which was 1.46 times higher than the control solution.

CONCLUSIONSThe SL-based vesicles obtained using high pressure homogenization method are characterized by small particle size, narrow distribution, good stability, and powerful permeation-enhancing effect, which enables them to be good carriers for the buccal delivery of insulin.

Absorption ; Administration, Topical ; Chemistry, Pharmaceutical ; Drug Carriers ; administration & dosage ; chemical synthesis ; pharmacokinetics ; Drug Delivery Systems ; methods ; Insulin ; metabolism ; Mouth Mucosa ; metabolism ; Nanotechnology ; methods ; Phosphatidylcholines ; pharmacokinetics ; Soybeans ; chemistry

OBJECTIVETo determine the location of the vertebral artery foramens from C(3) to C(6) and their relationship to the point 1 mm medial to the center of the lateral mass and to identify the value of oblique radiograph for cervical lateral mass screw trajectory by a cadaveric study.

METHODS(1) Twenty-eight cervical specimens (C(3)-C(7)) of human cadavers aged from 28 to 79 years were analysed. The transverse radiographs of C(3)-C(6) vertebrae were taken and the angle between the parasagittal plane and the line connecting the point of the lateral mass with the lateral limit of the transverse process foramen of C(3)-C(6) were measured. (2) The K-wires were drilled into lateral mass of C(3)-C(6) starting 1 mm medial to the center of the lateral mass and exiting by the juncture between the transverse process and the facet in ten specimens. Four wire placements under direct visualization, including placement of the wire tip staying the ventral cortex and 2, 4, 6 mm over-penetration of the ventral cortex of lateral mass, were performed separately on each specimen. After each placement, radiographs were taken on 45 degrees oblique left and 45 degrees oblique right views. Each intervertebral foramen on the oblique radiographs was divided into two parts: superior and inferior parts. The former is the true intervertebral foramen, while the latter is the intertransverse foramen on the gross specimen. The number of wire tips in each part was quantified for each placement. All results on the radiographs were compared with those on the gross anatomy.

RESULTS(1) The angles between the parasagittal plane and the line connecting the posterior starting point of the lateral mass with the lateral limit of the transverse foramen (C(3)-C(6)) were lateral to the sagittal plane, ranging from 5 degrees to 12 degrees. Among the vertebrae, there were no statistically significant difference (P > 0.05). (2) 15% of the wires without over-penetration and 41.3% with 2 mm over-penetration were found in the inferior parts of the intervertebral foramen in oblique views, while the wires were not noted in the intervertebral foramen by gross anatomy. with 4 mm over-penetration of the ventral cortex, 35% and 65% of wires were noted in the superior and inferior parts of the intervertebral foramen respectively, while only 28.8% of wires were found in the inferior part approximating the nerve roots in gross specimens. With 6 mm over-penetration, the number in the intervertebral foramen were 63.8% superiorly and 36.2% inferiorly on the oblique radiographs while all the tips were at the inferior part (intertransverse foramens) in gross specimens. The tip of wire crossed the line connecting the posterior borders of the intervertebral foramens in oblique radiographs when it penetrated the ventral cortex of lateral mass 4 mm or more.

CONCLUSIONS(1) There is no risk of damaging the vertebral artery if a screw is directed more than 15 degrees lateral to the sagittal plane at C(3 approximately 6) starting 1 mm medial to the center of the lateral mass. (2) Ideal screw tip position on oblique radiograph may not cross the line connecting the posterior borders of the intervertebral foramen on radiograph. If the screw tip is noted in the superior part of intervertebral foramen on the oblique radiograph, the screw may be identified as dangerous.

Adult ; Aged ; Bone Screws ; Cadaver ; Cervical Vertebrae ; anatomy & histology ; surgery ; Humans ; Internal Fixators ; Middle Aged ; Models, Anatomic ; Spinal Fusion ; instrumentation ; methods ; Spinal Nerve Roots ; anatomy & histology ; Vertebral Artery ; anatomy & histology

OBJECTIVETo investigate the effect of bone mesenchymal stem cell (BMSC) transplantation on repair of glomerular podocytes and on the Nephrin expression in rats with puromycin aminonucleoside (PAN) -induced nephrosis.

METHODSForty-five Sprague-Dawley rats were randomly divided into three groups (n=15 each): a nephrosis model group that received a single intraperitoneal injection of PAN (0.15 mg/g); a BMSC transplantation group that received a single intraperitoneal injection of PAN (0.15 mg/g) followed by BMSC transfusion; a control group that received a single intraperitoneal injection of normal saline. Ten days after injection, the rats were sacrificed. The 24 hrs urinary protein content and serum albumin and cholesterol levels were measured 24 hrs before sacrifice. Changes of glomerular podocytes were observed under an electron microscope. Brdu labeled positive cells in kidneys were measured by immunohistochemical technology. RT-PCR and Western blot were used to assess the expression of mRNA and protein of Nephrin.

RESULTSIn the nephrosis model group, urinary protein and blood cholesterol contents increased, plasma albumin content decreased compared with those in the control group. Extensive fusion of podocyte foot processes was observed in the nephrosis model group. The BMSC transplantation group had decreased urinary protein and blood cholesterol contents and increased plasma albumin content compared with the nephrosis model group. Fusion of podocyte foot processes was also improved. Brdu labeled positive cells were seen in kidneys in the BMSC transplantation group, but not in the nephrosis model and the control groups. Nephrin mRNA and protein expression decreased significantly in the nephrosis model group compared with that in the control group. The BMSC transplantation group had increased Nephrin mRNA and protein expression compared with the nephrosis model group.

CONCLUSIONSBMSCs can repair glomerular podocytes in PAN-induced nephrosis rats, and the changes of Nephrin expression may be involved in the process.

Animals ; Bromodeoxyuridine ; metabolism ; Kidney ; pathology ; Male ; Membrane Proteins ; genetics ; Mesenchymal Stem Cell Transplantation ; Nephrosis, Lipoid ; chemically induced ; pathology ; therapy ; Podocytes ; pathology ; Puromycin Aminonucleoside ; toxicity ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction

Objective: To evaluate the immunogenicity of HIV-1 clade C/B’ vaccine based on modified vaccinia virus Ankara (MVA) vector in mice. Methods: Mice were inoculated with 3-dose HIV vaccine by intramuscular injection. Blood sample were collected every second week, and then the antibodies against HIV were detected. At week 6, mice were killed and cellular immune responses were examined by ELISPOT. Result: The number of spot forming cells in the 107 pfu/ml -dose group was more than those of 105 pfu/ml -dose and 106 pfu/ml -dose groups significantly. HIV specific antibodies emerged at week 2 and elevated rapidly at week 4 and week 6. The level of specific IgG in the 107 pfu/ml -dose group was more than those of 105 pfu/ml -dose and 106 pfu/ml -dose groups significantly. Conclusion: The ADMVA induces both humoral immunoresponse and cellular immune responses.

OBJECTIVETo develop a system to rescue virus by intracellular expression of T7 RNA Polymerase.

METHODSThe gene of T7 RNA Polymerase was amplified and cloned to VR1012 by molecular biological technology. The expression plasmid VR-1a was then identified. VR-1a and EV71 infectious plasmid were co-transfected in Vero cell. CPE was observed and viral gene viral antigen were detected.

RESULTSThe gene of T7 RNA Polymerase was successfully cloned into vector VR1012. Vero cell developed to CPE after being transfected VR-1a and EV71 infectious plasmid. EV71 gene was amplified by RT-PCR from the culture. EV71 antigen was also detected by ELISA.

CONCLUSIONThe method can be used to rescue virus. It could apply to immunologic research of EV71 DNA vaccine.

Animals ; Cercopithecus aethiops ; DNA-Directed RNA Polymerases ; genetics ; metabolism ; Enterovirus A, Human ; genetics ; physiology ; Gene Expression ; Genetic Engineering ; methods ; Genetic Vectors ; genetics ; metabolism ; HeLa Cells ; Humans ; Plasmids ; genetics ; metabolism ; Transfection ; Vero Cells ; Viral Proteins ; genetics ; metabolism ; Virus Replication

Objective of this study was to evaluate the efficacy and safety of haploidentical or unrelated donor hematopoietic stem cell transplantation (HSCT) for patients with severe aplastic anemia (SAA). Twenty patients with SAA received allogeneic HSCT from haploidentical or unrelated donors (14 from haploidentical donors and 6 from unrelated donors) from November 2005 to May 2011. Conditioning regimen consisted of fludarabine (FLU), cyclophosphamide (Cy) and anti-thymocyte immunoglobulin (ATG). The patients were administrated with G-CSF-primed bone marrow and mobilized peripheral blood as grafts from haploidentical donor or only mobilized peripheral blood from the unrelated donor. The results showed that the median time of neutrophil and platelet engraftment were 14 (11 - 20) d and 17 (13 - 31) d respectively. All patients who achieved engraftment had complete hematologic recovery with complete donor chimerism, except for two patients who developed graft failure in 2 months after transplantation. Four cases developed acute grade IIGVHD. The chronic GVHD occurred in 7 of the 16 evaluable cases (6 limited, 1 extensive). 14 patients got disease-free survival with follow-up to January 2012. The disease-free survival rate was 68.9%. It is concluded that the haploidentical or unrelated donor hematopoietic stem cell transplantation may become a viable therapeutic option for severe aplastic anemia patients who lack suitable human leukocyte antigen-matched donors and fail immunosuppressive therapy.
Adolescent ; Adult ; Anemia, Aplastic ; surgery ; Child ; Child, Preschool ; Female ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Immunosuppressive Agents ; therapeutic use ; Male ; Transplantation Conditioning ; methods ; Transplantation, Homologous ; Unrelated Donors

This study was aimed to explore the distribution characteristics of the human platelet antigen (HPA) gene of human platelet donors and its polymorphism in Mudanjiang area of Heilongjiang Province in China, to determine platelet antigen system with clinical significance by judging the rate of incompatibility of HPA, as well as to establish a database of donors' HPA. The genotyping of 154 unrelated platelet donors was performed by means of PCR-SSP. The frequencies of gene and genotype were calculated and compared with that in other areas. The results showed that the genes 1a-17a of HPA-a were all expressed in the 154 healthy and unrelated platelet donors. Only genes 1b, 2b, 3b, 5b, 6b and 15b of HPA-b were expressed while genes 4b, 7b-14b, 16b were not expressed. Among the genotypes, aa homozygosity was predominant and HPA15 had the greatest heterozygosity, while HPA3 had lower heterozygosity. There were 23 combined types of HPA, 5 of them had a rate higher than 10%, and the frequencies of the other 18 were lower than 8%. HPA genotype frequencies showed a good consistency to Hardy-Weinberg equilibrium. It is concluded that the distribution of the allele polymorphism of HPA1-HPA17 in Mudanjiang area has its own characteristics, compared with other areas and some countries, the local HPA genotype database of platelet donors is established in Mudanjiang area, which can provide the matching donors for clinical use with immunological significance.
Adolescent ; Adult ; Alleles ; Antigens, Human Platelet ; genetics ; Blood Donors ; China ; Databases, Genetic ; Female ; Gene Frequency ; Genotype ; Heterozygote ; Homozygote ; Humans ; Male ; Middle Aged ; Young Adult