Objective:To produce rhBMP-4 with bioactivity in E.coli. Methods: The full-length human BMP-4 gene was mutated by PCR without changes in amino acid sequence, then the synthesized gene was cloned into plasmid pET-3c, transducted into BL21(DE)plysS, and induced by adding IPTG to a final concentration of 1.0 mmol/L. The protein product was purified using ion-exchange chromatography method and then renaturated, bioactivity was checked by C2C12 differentiation in vitro and mouse ectopic bone formation in vivo. Results: A 438 bp gene fragment encoding mature peptide of hBMP-4 was cloned , the protein product was mostly in the form of inclusion body, after renaturation, the engineering protein shows better bioactivity. Conclusion:The mutant strategy can enhance the expression of bioactive rhBMP-4 in E.coli expression system.

Adolescent ; Adult ; Aged ; Brain ; blood supply ; diagnostic imaging ; Carbon Monoxide Poisoning ; diagnostic imaging ; Child ; Female ; Humans ; Male ; Middle Aged ; Tomography, Emission-Computed, Single-Photon

BACKGROUNDA bioactive compound from Paecilomyces tenuipes (BCPT) has an inhibitory effect on monoamine oxidase A (MAO-A) and monoamine oxidase B (MAO-B) in vitro and in vivo, which indicates BCPT may be a potential antidepressant. In this study we aimed to study the antidepressant effects of BCPT in the chronic unpredictable stress (CUS) model in rats and explore underlying mechanisms in the hypothalamic-pituitary-adrenal (HPA) axis.

METHODSThe antidepressant effects of BCPT were studied in the chronic unpredictable stress model in rats. Animals were housed isolated, except the control group. Rats were exposed daily to different random stressors from day 1 to 21. Awarding response was detected by calculating the 24-hour consumption of sucrose water. Cortisol (CORT) and adrenocorticotropic hormone (ATCH) contents in serum and arginine vasopressin (AVP) contents in the pituitary body were detected by radio immunoassays. Total RNA of hippocampus or hypothalamus was extracted and subjected to reverse transcription-polymerase chain reaction (RT-PCR) for the measurement of corticotrophin releasing hormone (CRH) mRNA or mineralocorticoid receptor (MR) mRNA and glucocorticoid receptor (GR) mRNA levels. Statistical analyses were performed using one way analysis of variance (ANOVA) followed by Student-Newman-Keuls (SNK) test.

RESULTSChronic unpredictable stress resulted in reduction of sensitivity to reward and abnormality in the HPA axis in the animal model. BCPT improved the reward reaction as measured by increasing sucrose consumption, remarkably reduced serum CORT and ACTH levels and the AVP content in the pituitary body in the CUS-treated rats, decreased the expression of CRH mRNA, enhanced the expression of hippocampus MR mRNA, GR mRNA and decreased the ratio of MR/GR.

CONCLUSIONSBCPT has potentially antidepressant-like activity and normalized the HPA axis hyperactivity in a CUS model of depression in rats. This may be an important mechanism of its antidepressant effect.

Animals ; Antidepressive Agents ; pharmacology ; Chronic Disease ; Corticotropin-Releasing Hormone ; genetics ; Hydrocortisone ; blood ; Hypothalamo-Hypophyseal System ; drug effects ; physiology ; Male ; Monoamine Oxidase Inhibitors ; pharmacology ; Paecilomyces ; chemistry ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Receptors, Glucocorticoid ; genetics ; Receptors, Mineralocorticoid ; genetics ; Stress, Psychological ; physiopathology ; Sucrose ; administration & dosage

Osteoarthritis is a degenerative joint disease,with the characters of degradation of articular cartilage, the formation of the joint marginal osteophyte and synovium lesions. Previous studies have focused on the treatment of articular cartilage lesions. In recent years, new research in shows synovial inflammation plays an important role in OA. Synovium lesions and synovial inflammation-related factors induced the degradation and destruction of articular cartilage, and promoted the development of osteoarthritis. The role of synovial lesions in osteoarthritis is increasingly prominent, and the treatment for synovial lesions will become a new target. So this paper reviews the various manifestations of synovial in osteoarthritis.
Humans ; Osteoarthritis ; pathology ; Synovial Membrane ; pathology

OBJECTIVETo observe the change of cytokine interleukin IL-1 beta, IL-6, IL-10, tumor necrosis factor-alpha (TNF-alpha) occurred in acute paraquat (PQ) poisoning rats and to investigate the mechanism of acute lung injury caused by paraquat (PQ) poisoning.

METHODSAll 72 healthy adult Wistar rats were random assigned into normal control groups, paraquat high dose group (120 mg/kg), paraquat middle dose (60 mg/kg) group, paraquat low dose group (30 mg/kg). Three observing periods of time included 8, 24, 72 h and the standards of TNF-alpha, IL-1 beta, IL-6, IL-10 were determined.

RESULTSEvery index of the PQ group was significantly higher than that in the NS group at the same period of time (P<0.05 or P<0.01). In the 72 h group, the high dose group was significantly higher than the middle and low dose group (P<0.05), and there was no significantly difference between the middle and low dose group (P>0.05). For the comparison of index in the same dose group, the group of 72 h was much higher than 8 h group and 24 h group (P<0.05), and there was no difference between the 8h group and 24 h group (P>0.05).

CONCLUSIONThe cytokine may play an important role in paraquat-induced acute lung tissue injury.

Acute Disease ; Animals ; Cytokines ; blood ; Disease Models, Animal ; Female ; Interleukin-10 ; blood ; Interleukin-1beta ; blood ; Interleukin-6 ; blood ; Male ; Paraquat ; poisoning ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; blood

Objective To compare the efficacy of surgical treatment(ST) and mechanical ventilation treatment(MVT) for flail chest.Methods From March 2013 to May 2017,81 patients with flail chest who underwent ST(28 cases) and MVT(53 cases) were enrolled in ICU.The relevant indicators of efficacy were compared including mechanical ventilation (MV) time,bed rest time,oxygen administration time,catheterization time,antibiotic use time,drug analgesia,drug analgesia patients,proportion of stop/postoperative sputum alone,ICU treatment time,hospital stay,mortality,readmission within two months and long-term pulmonary function after discharge.Results The MV time,bed time,oxygen time,antibiotic use time,ICU treatment time and hospitalization time of MVT were longer than those of ST group,the differences were significant(P ＜ 0.05).MVT had more analgesic times,more analgesic cases,smaller proportion of offline/postoperative sputum along and less hospitalization fees compared with ST group,the differences were significant(P ＜0.05).There was no significant difference in the main indexes of pulmonary function between two groups in 3 months after discharge.Conclusion ST and MVT respectively has advantages and disadvantages for flail chest.There is no significant difference in long-term pulmonary function of ST and MVT.While MVT has less trauma and lower cost.The appropriate treatment could be selected according to the clinical situation.

FGF21 (fibroblast growth factor 21) is a recently described member of the FGF family. It has been previously demonstrated that FGF21 is a potent regulator of glucose homeostasis. To improve stability of FGF21 for better efficacy, a new form of recombinant FGF21 was generated by fusion of a full length FGF21 gene and the Fc fragment of human IgG4 with flexible linker sequence. To examine the glucose regulation activity of FGF21-L-Fc, 3T3-L1 pre-adipocytes were differentiated into adipocytes, and glucose uptake activity of FGF21-L-Fc was examined by glucose oxidase and peroxidase (GOD-POD) assay. The results showed that in comparison with wild type FGF21, FGF21-L-Fc was more potent in stimulation of glucose uptake by 3T3-L1. In vivo studies on the modified protein demonstrated that FGF-L-Fc had a better efficacy in lowering blood glucose of the STZ-induced diabetic animals and controlled glucose level for a longer time. The results provided a sound basis for further studies.
3T3-L1 Cells ; Adipocytes ; metabolism ; Animals ; Blood Glucose ; metabolism ; Diabetes Mellitus, Experimental ; drug therapy ; metabolism ; Escherichia coli ; metabolism ; Fibroblast Growth Factors ; genetics ; pharmacology ; Glucose ; metabolism ; Immunoglobulin G ; genetics ; pharmacology ; Male ; Mice ; Recombinant Proteins ; genetics ; pharmacology

OBJECTIVETo characterize the human immunodeficiency virus (HIV) -specific T lymphocyte responses and identify the immunodominant regions in Chinese HIV-1 recombinant subtype B/C chronic infectors at complete genome level.

METHODSTwenty-five HIV-1B/C recombinant chronic infectors were screened for their specific T lymphocyte responses to a panel of peptides corresponding to the complete HIV-1 subtype B genome by gamma interferon ELISPOT assay. Kruskal-Wallis nonparametric analysis of variance was used to test significant differences across gene regions, and Tukey pairwise analysis was used to identify differences between gene regions. Spearman rank correlation was used to assess the relation between responses. Results The order of recognized frequencies of specific T lymphocyte responses to HIV proteins was Nef>Vpr>Gag>Pol>Vpu>Env>Rev>Vif>Tat. When adjusted for protein length, Nef, Vpr, Gag, and Pol were the most intensely targeted proteins and the central region of Nef, Gag p24, Pol RT, and Vpr was most frequently recognized. No significant correlation was observed between the magnitude of IFN-gamma production of HIV-l-specific T lymphocyte responses and plasma viremia, breadth of response and CD4 counts. Conclusion The central region of Nef, Gag p24, Pol RT, and Vpr is most frequently targeted in HIV-1 B/C recombinants chronic infectors. HIV-l-specific T lymphocyte responses and plasma viremia or CD4 counts play no protective role at complete genome level in these infectors.

Adolescent ; Adult ; Asian Continental Ancestry Group ; CD4 Lymphocyte Count ; Chronic Disease ; Female ; HIV Infections ; immunology ; HIV-1 ; immunology ; Human Immunodeficiency Virus Proteins ; Humans ; Male ; T-Lymphocytes ; physiology ; Viral Load ; Young Adult

OBJECTIVETo test the hypothesis that the introduction of antisense transforming growth factor beta receptor I (TBRI) plasmid and antisense tissue inhibitor of matrix metalloproteinase (TIMP-1) eukaryotic expressing plasmid into a rat liver fibrosis model may influence the progression of liver fibrosis.

METHODSFragments of TBRI cDNA and TIMP-1 cDNA were obtained by reverse transcription polymerase chain reaction (RT-PCR) and then amplified by nest PCR. pcDNA3.1(+)-antisense TBRI eukaryotic expressing plasmid was constructed by directional and inverted joins with the purified linear pcDNA3.1(+) and the purified fragment of TBRI, as well as, pcDNA3.1(+)-antisense TIMP-1 eukaryotic expressing plasmid. The recombinant was identified by restriction endonuclease digestion and DNA sequence analysis. The recombinant plasmids were encapsulated with Lipofectmine 2000, and then they were injected intraperitoneally into the liver fibrosis model rats. The protein expression of type I collagen was evaluated by immunohistochemistry. VG staining of liver slides of the rats was used for histopathological examination.

RESULTSCompared with the empty plasmid control group and the disease control group, the deposition of type I collagen decreased in the three antisense treatment groups: antisense TBRI group (4.37+/-1.30) x 10(5), P less than 0.05; antisense TIMP-1 group (3.40+/-0.91) x 10(5), probability value less than 0.05; antisense TBRI + antisense TIMP-1 group (0.90+/-0.32) x 10(5), P less than 0.01; treatment control group (6.90+/-1.61) x 10(5); disease control group (7.34+/-1.68) x 10(5); and the normal control group (0.41+/-0.21) x 10(5)]. Significant differences in the pathological grades of fibrosis were found between the normal control group and the other five groups (P less than 0.05) and also between the disease control group and the three antisense treatment groups (antisense TBRI group P less than 0.05; antisense TIMP-1 group P less than 0.05; antisense TBRI + antisense TIMP-1 group P less than 0.01), but no difference was found between the empty plasmid control group and disease control group (P more than 0.05).

CONCLUSIONBoth antisense TBRI eukaryotic expressing plasmid and antisense TIMP-1 eukaryotic expressing plasmid can inhibit the progress of liver fibrosis. A combined action can inhibit the progress of liver fibrosis more.

Animals ; Antisense Elements (Genetics) ; Female ; Genetic Vectors ; Liver Cirrhosis ; pathology ; Protein-Serine-Threonine Kinases ; genetics ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Receptors, Transforming Growth Factor beta ; genetics ; Tissue Inhibitor of Metalloproteinase-1 ; genetics

OBJECTIVETo explore the pathological mechanisms of Guizhi Decoction () syndrome and the therapeutic molecular mechanisms of the Guizhi Decoction, Mahuang Decoction (), Sangju Decoction ( ) and Yinqiao Powder (), as well as the potentially biological basis that Guizhi Decoction is most effective only for the patients with Guizhi Decoction syndrome in clinical practice.

METHODSWe first got serum samples from the patients suffering from both upper respiratory tract infection and Guizhi Decoction syndrome identified by the doctors of Chinese medicine (CM) in the clinic. Four formulas with therapeutic actions of pungent warmth or pungent coolness for superficial syndromes were chosen and four kinds of rat serum samples each containing one of the above-mentioned herbal formulas were collected, then the effects of Guizhi Decoction syndromes' patient serum as well as the effects of sera containing the formulas after being stimulated by the patient serum samples on both the mRNA expression of certain toll-like receptor (TLR) subtypes and the release of some inflammatory cytokines in RAW264.7 cells were tested and analyzed in vitro.

RESULTSThe expression of TLR-3, TLR-4 and TLR-9 mRNA among the 9 tested TLR subforms were up-regulated in the macrophages stimulated by the sera from untreated upper respiratory infection patients with the Guizhi Decoction syndrome (symptomcomplex). The products such as interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α and interferon (IFN)-β from stimulated macrophages through TLR signaling pathways were also increased correspondingly. Interestingly, the changes induced by the Guizhi Decoction syndrome patients' sera were masked significantly after the macrophages were incubated with the sera from donors treated with Guizhi Decoction. Similarly, the three other exterior-releasing formulas were all effective in reversing the up-regulated changes of certain TLR subforms to different degrees, but both the number of targeted TLRs and efficacy of them seemed to be inferior to that of Guizhi Decoction.

CONCLUSIONEvidence from these experiments might contribute to the scientific explanation of both the pharmacological mechanisms of Guizhi Decoction and also the CM theory that Guizhi Decoction is specifically prescribed for the treatment of Guizhi Decoction syndrome (The gearing formula to the symptom-complex).

Animals ; Cell Survival ; drug effects ; genetics ; Cytokines ; secretion ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gene Expression Regulation ; drug effects ; Healthy Volunteers ; Humans ; Inflammation Mediators ; metabolism ; Inhibitory Concentration 50 ; Macrophages ; drug effects ; metabolism ; Male ; Mice ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Syndrome ; Toll-Like Receptors ; genetics ; metabolism