OBJECTIVETo study the overexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells for repairing articular cartilage injury in vivo.

METHODSRabbit bone marrow mesenchymal stem cells (BMSCs) were transduced with lentivirus vector containing Sox9 gene and then cartilage specific molecule was detected by RT-PCR in vitro. Total 48 knee joints of 24 mature New Zealand white rabbits were randomly divided into 3 groups according to different defect treatment. After animals anesthesia,a full-thickness cylindrical cartilage defect of 4 mm diameter and 3 mm deep was created in the patellar groove using a stainlesssteel punch. Meanwhile, the transfected cells were implanted to repair the rabbit model with full-thickness cartilage defects. Cartilage defects tissue was observed with light microscope, electron microscope, HE and immunohistochemistry staining to assess the repair of defects by the complex at 6 weeks or 12 weeks after the implantation.

RESULTSAt 3 days after the transfection, Sox9 gene expression was highest and Sox9 gene expression decreased with the increase of time. At 3 days after the transfection, the expression of collagen type II began and reached the peak at 14 days. It showed that the bone marrow mesenchymal stem cells went into chondrogenic differentiation after transfected by Sox9 gene. Histological observation showed that at 6 weeks after the operation, the defects in the experimental group was filled with hyaline like cartilage tissue, 12 weeks after operation,the defects of cartilage and subchondral bone had satisfactory healing. Both at 6 and 12 weeks postoperatively, the defects were filled with fibrous tissues in control groups. Meanwhile, immunohistochemical staining of sections with type II collagen antibodies showed the proteins in the regenerated tissue stained positive for type II collagen and stronger than the control groups. The histological scoring system indicated that the cartilage repair of experiment groups were better than the two control groups with statistical significances.

CONCLUSIONOverexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells (BMSCs) promote the repair of cartilage defect.

Animals ; Bone Marrow Cells ; metabolism ; Bone Marrow Transplantation ; Cartilage, Articular ; injuries ; metabolism ; Cell- and Tissue-Based Therapy ; Female ; Genetic Vectors ; genetics ; metabolism ; Humans ; Lentivirus ; genetics ; metabolism ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; metabolism ; Osteoarthritis ; genetics ; metabolism ; therapy ; Rabbits ; SOX9 Transcription Factor ; genetics ; metabolism ; Tissue Engineering

OBJECTIVETo observe the changes of systemic and pulmonary hemodynamics and the plasma levels of inducible nitric oxide synthase (iNOS) and endothelin-1 (ET-1) and investigate their association in patients with hepatopulmonary syndrome (HPS).

METHODSTwenty-six patients with HPS undergoing orthotopic liver transplantation (OLT) were enrolled in this study with 20 patients without hypoxemia as the control group. Blood samples were taken one day before OLT to measure the plasma levels of iNOS and ET-1 using fluorescence quantitative polymerase chain reaction (FQ-PCR) and radioimmunoassay, respectively, with 10 healthy volunteers serving as the healthy control group. Before the operation for OLT, the parameters of systemic and pulmonary hemodynamics were monitored after anesthesia induction.

RESULTSThe systemic and pulmonary hemodynamics in patients without hypoxemia was characterized by high cardiac output and low resistance, and by comparison, the patients with HPS showed even higher cardiac output and lower mean pulmonary artery pressure, pulmonary artery wedge pressure, systemic vascular resistance and pulmonary vascular resistance. The two patient groups had comparable plasma iNOS and ET-1 levels, which were both higher than those in the healthy control group.

CONCLUSIONThe hemodynamics in patients with end-stage liver disease exhibit a pattern of high cardiac output and low resistance, which is more obvious in HPS patients possibly in association with elevated plasma levels of iNOS and ET-1.

Adult ; Aged ; Case-Control Studies ; Endothelin-1 ; blood ; Female ; Hemodynamics ; physiology ; Hepatopulmonary Syndrome ; blood ; physiopathology ; Humans ; Male ; Middle Aged ; Nitric Oxide Synthase Type II ; blood ; Pulmonary Circulation ; physiology ; Young Adult

OBJECTIVETo establish quality standard of Zhuang medicine Lonicera dasystyla, and provide scientific basis for the quality control of L. dasystyla.

METHODCharacteristics of materia medica, microscopic features, TLC indentification, inspection, extractum and determination of chlorogenic acid, macranthoidin B, dipsacoside B were carried out through the experience, microscopic, physical and chemical methods, respectively. The standard of quality control was formulated thereafter.

RESULTThe characteristics of materia medica, microscopic features, TLC indentification were specified, the average contents of water, total ash, acid-insoluble ash, alcohol-soluble extracts, chlorogenic acid were 11.6%, 6.6%, 0.2% , 24.4%, 1.16%, respectively, the total amount of macranthoidin B and dipsacoside B was 3.13%. Quality standard of L. dasystyla was proposed according to experimental results.

CONCLUSIONThe quality of L. dasystyla can be controlled effectively with the quality standard.

Chlorogenic Acid ; analysis ; isolation & purification ; Chromatography, Thin Layer ; Drugs, Chinese Herbal ; chemistry ; standards ; Lonicera ; chemistry ; Oleanolic Acid ; analogs & derivatives ; analysis ; isolation & purification ; Quality Control ; Saponins ; analysis ; isolation & purification ; Solubility

OBJECTIVETo investigate the role of sodium cromoglycate in brain protection and its effects on brain tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) expressions after global cerebral ischemia-reperfusion (IR) injury in gerbils.

METHODSTwenty-four healthy male gerbils were randomized into 3 equal groups, namely the sham-operated group with isolation of the bilateral carotid arteries but without occlusion, IR injury model group with bilateral carotid artery occlusion, and sodium cromoglycate treatment group with bilateral carotid artery occlusion and sodium cromoglycate administration at 25 mg/kg via the lingual vein as soon as the reperfusion start with another dose 1 h later. The animals were then sacrificed and the thalamus were removed, fixed in 10% formaldehyde and sliced for observation under light microscope with HE staining. The rest brain tissues were prepared into homogenate to determine the content of TNF-alpha and IL-1beta. The right hemispheres of the gerbils were measured for wet weight and dry weight to calculate the water content in the brain.

RESULTSThe water content in the brain of the gerbils in the model group was the highest among the groups, and that in sodium cromoglycate treatment group was significantly less than that of the model group (P<0.05). Microscopic examination showed the most severe brain tissue damage in the model group with also the highest TNF-alpha and IL-1beta levels in the brain. The brain TNF-alpha and IL-1beta levels in sodium cromoglycate group were significantly decreased as compared with those in the model group (P<0.05).

CONCLUSIONSodium cromoglycate can alleviate brain IR injury possibly by lowering the TNF-alpha and IL-1beta levels in the brain tissues.

Animals ; Brain Ischemia ; metabolism ; Cromolyn Sodium ; pharmacology ; Gerbillinae ; Interleukin-1beta ; metabolism ; Male ; Neuroprotective Agents ; pharmacology ; Random Allocation ; Reperfusion Injury ; prevention & control ; Tumor Necrosis Factor-alpha ; metabolism

AIMTo explore the changes of rat platelet phospholipase A2 (PLA2) mRNA content in bacteria infected rat and study the cDNA and amino acid sequences of the PLA2 structure to lay a good foundation for the development of new antibiotics.

METHODSThe PLA2 mRNA level in blood was determined by RT-PCR. The DNA sequence was cloned and analyzed.

RESULTSAfter injection of bacteria in rats, the mRNA level of PLA2 in blood increased markedly. The cDNA and amino acid sequence were highly homologous to other PLA2 cDNA from different tissues of the rat.

CONCLUSIONPlatelet PLA2 in blood responded quickly to bacteria infection in gene level. Therefore, the PLA2 protein was produced increasingly which was shown to control the infection with bacteria. Although there are little difference between PLA2 cDNA cloned from blood and other sources in DNA and amino acid sequences, the catalytic site for enzymatic activity and basic structure are identical.

Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; DNA, Complementary ; metabolism ; Male ; Molecular Sequence Data ; Phospholipases A ; blood ; genetics ; metabolism ; Phospholipases A2 ; RNA, Messenger ; metabolism ; Rats ; Rats, Wistar ; Staphylococcal Infections ; blood ; Staphylococcus aureus

Objective To explore the method of pharmaceutical care for patients with non-ST segment elevation acute myocardial infarction and pulmonary tuberculosis by clinical pharmacist.Methods Clinical pharmacist participated in an acute non-ST segment elevation myocardial infarction patients with pulmonary tuberculosis,analyzing the treatment options for patients,assessing the efficacy of anti-platelet drugs,discussing gene polymorphisms and the interactions efficacy between anti-TB drugs and anti-platelet drugs.Results and conclusion Analyzing the function and characteristics of each drug,managing of drug interactions,providing individual treatment strategies for patientssare the key points for clinical pharmacists participating in clinical work.

Objective:To study the overexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells for re?pairing articular cartilage injury in vivo. Methods:Rabbit bone marrow mesenchymal stem cells(BMSCs)were transduced with lentivirus vector containing Sox9 gene and then cartilage specific molecule was detected by RT-PCR in vitro. Total 48 knee joints of 24 mature New Zealand white rabbits were randomly divided into 3 groups according to different defect treat?ment. After animals anesthesia,a full thickness cylindrical cartilage defect of 4 mm diameter and 3 mm deep was created in the patellar groove using a stainlesssteel punch. Meanwhile,the transfected cells were implanted to repair the rabbit model with full thickness cartilage defects. Cartilage defects tissue was observed with light microscope,electron microscope,HE and im?munohistochemistry staining to assess the repair of defects by the complex at 6 weeks or 12 weeks after the implantation. Re?sults:At 3 days after the transfection,Sox9 gene expression was highest and Sox9 gene expression decreased with the increase of time. At 3 days after the transfection,the expression of collagen typeⅡbegan and reached the peak at 14 days. It showed that the bone marrow mesenchymal stem cells went into chondrogenic differentiation after transfected by Sox 9 gene. Histologi?cal observation showed that at 6 weeks after the operation,the defects in the experimental group was filled with hyaline like cartilage tissue,12 weeks after operation,the defects of cartilage and subchondral bone had satisfactory healing. Both at 6 and 12 weeks postoperatively,the defects were filled with fibrous tissues in control groups. Meanwhile,immunohistochemical stain?ing of sections with typeⅡcollagen antibodies showed the proteins in the regenerated tissue stained positive for typeⅡcolla?gen and stronger than the control groups. The histological scoring system indicated that the cartilage repair of experiment groups were better than the two control groups with statistical significances. Conclusion:Overexpression of Sox9 gene on rab? bit bone marrow mesenchymal stem cells(BMSCs)promote the repair of cartilage defect.

OBJECTIVETo assess the prevalence of several tyrosine kinases (TKs) gene mutations including c-Kit, FLT3 and JAK2 V617F in core binding factor related acute myeloid leukemia (CBF-AML), and analyze their impact on clinical characteristics and prognosis.

METHODSMutations of c-Kit, FLT3-ITD and FLT3-TKD were detected by genomic DNA PCR and sequencing, and JAK2 V617F mutation screening by allele-specific PCR in 58 newly diagnosed CBF-AML patients [28 AML with inv(16) and 30 with t(8;21)], and analyze the patients clinical characteristics and prognoses.

RESULTSc-Kit aberrations were detected in 32.8% cases, including 6 cases mutated in exon 8 (mutKIT8) and 13 mutated in exon 17 (mutKIT17). MutKIT8 was more prominent in inv(16) than in t(8;21) patients (21.4% vs 0, P = 0.009). Only 2 cases had FLT3-ITD and 7 (12.1%) FLT3-TKD mutations. The result of JAK2 V617F mutation screenings in these CBF-AML patients was negative. The frequency of receptor tyrosine kinases(RTK) mutations was 46.6% and only one case had two kinds of missense mutations (mutKIT8 & TKD(+)). Median age of onset was higher for mutKIT17 than for wide-type c-Kit (wtKIT) patients (55 vs 31, P = 0.003). c-Kit mutations were significantly associated with decreased overall survival (OS) and continuous complete remission (CCR) rates (P = 0.053, and 0.048 respectively), and so did more for exon17 mutated patients reduced (P = 0.005, and 0.013 respectively). FLT3-TKD mutation showed no effects on prognosis of CBF-AML patients.

CONCLUSIONSRTK mutations are common in patients with CBF-AML. c-Kit mutations frequently and JAK2V617F mutation rarely appear in CBF-AML. c-Kit mutations, especially mutKIT17 confers higher relapse risk and poorer prognosis.

Adolescent ; Adult ; Aged ; Core Binding Factors ; DNA Mutational Analysis ; Female ; Humans ; Janus Kinase 2 ; genetics ; Leukemia, Myeloid, Acute ; diagnosis ; etiology ; genetics ; Male ; Middle Aged ; Mutation ; Prognosis ; Protein-Tyrosine Kinases ; genetics ; Proto-Oncogene Proteins c-kit ; genetics ; Young Adult ; fms-Like Tyrosine Kinase 3 ; genetics

OBJECTIVETo investigate the expression of several homeobox genes during the wound healing in fetal and adult skin and their roles in fetal scarless wound healing.

METHODSThe expressions of PRX-2, HOXB13, HOX2.2 and HOX2.3 during wound healing in fetal and adult skin were determined with in situ hybridization.

RESULTS(1) PRX-2 positive expression could be identified in normal fetal and adult skin, especially in the fetus. But there was difference in location sites of the genes. The positive expression in normal fetal skin was mainly found in the peripheral cells at the hair shafts within dermal papilla layers and was also found in the epithelium. Nevertheless, weak positive expression of PRX-2 was found in the epithelial basal layer cells in normal adult skin but not in dermal tissue. There was strong positive expression of the PRX-2 in the tissue around the wound in fetus but not of that in adults except the epithelial basal layers. (2) Positive expression of HOXB13 could be identified in both normal fetal and adult skins. And the expression was concentrated mainly in hair follicle cells in the dermis and in the basal layer cells in the epithelium. Furthermore, the expression became weak after trauma, especially in fetal skin. (3) The positive expression of HOX2.2 and HOX2.3 in normal fetal skin was observed mainly in the whole layer of the epithelium and especially in the epithelial basal layers. Weak positive expression could be found in the dermis and strong expression found in the tissue near the wound. But there was no positive expression of the HOX genes in normal adult skin and wounds.

CONCLUSIONThe difference in the HOX expression in fetal and adult skin wound healing might be the key factor leading to different wound healing. Homeobox genes might be closely related with the developmental biology.

Adolescent ; Adult ; Animals ; Burns ; pathology ; Child ; Female ; Fetus ; Gene Expression ; Genes, Homeobox ; Homeodomain Proteins ; metabolism ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Skin ; embryology ; Wound Healing ; genetics ; Young Adult

OBJECTIVETo investigate the feasibility of monitoring therapeutic effect of adenovirus vector containing IL12-IRES-CKb gene on a rabbit VX2 liver tumor model by using phosphorous-31 magnetic resonance spectroscopy (31P MRS).

METHODSA total of 18 healthy New Zealand White rabbits were used to generate animal models by implanting VX2 tumor chips into livers through laparotomy. Tumor-bearing animals were randomly divided into three groups and were injected with AdCMVIL12-IRES-CKb, AdCMV-Empty and saline respectively via ear veins. 31P MRS scan was performed after animals were fed with creatine solution for five days. Animals were euthanized thereafter and tumors were removed for pathological examination, immunohistochemistry (IHC) staining and protein analysis (Western blot).

RESULTSThe intrahepatic and seral expressions of creatine kinase (CKb) and IL-12 were detected only in AdCMVIL12-IRES-CKb group. Tumor diameters pre- and post- treatment in three groups were 1.63+/-0.04 vs 1.62+/-0.03 in AdCMVIL12-IRES-CKb group (P = 0.229), 1.59+/-0.05 vs 1.84+/-0.11 in AdCMV-Empty group (P = 0.003) and 1.60+/-0.02 vs 2.07+/-0.12 in saline group (P = 0.001), respectively. Pcr Changes between pre- and post- treatment among the three groups were compared (F = 6.235, P value is less than 0.05). PCr increased significantly in AdCMVIL12-IRES-CKb group as compared to AdCMV-Empty (P = 0.004) and saline group (P = 0.049), whereas no change found between AdCMV-Empty and saline group (P = 0.153).

CONCLUSION31P MRS, an effective and non-invasive functional imaging method, can be used to monitor the therapeutic effect of adenovirus vector containing IL12-IRES-CKb gene on rabbit VX2 liver tumor model through detecting metabolic product of imaging reporter gene CKb (pCr).

Adenoviridae ; genetics ; Animals ; Creatine Kinase ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; Interleukin-12 ; genetics ; Liver Neoplasms, Experimental ; genetics ; pathology ; Magnetic Resonance Spectroscopy ; Rabbits