OBJECTIVE: To investigate the protective effects of metformin hydrochloride (MF) on osteoblasts which were exposed to constant high glucose condition, and the relationship between MF and the expression of peroxisome proliferator-activated receptor (PPAR)γ. METHODS: Mouse cranium osteoblasts were cultured in vitro and divided into 5.5 mmol · L-1 normal glucose +0 μmol · L-1 MF group, 25 mmol · L-1 high glucose with different concentration of MF (0, 25, 50, 100 μmol · L-1 MF, respectively) and intervened for 1,2 and 3 w, then the mRNA expression levels of PPARγ and bone morphogenetic protein (BMP)-2 were detected by RT-PCR, and the protein expression levels of PPARγ was examined by Western blot. RESULTS (1)The mRNA expression levels of PPARγ was marked higher in high glucose group compared to the control group with the same intervene time (all P < 0.05); when the intervention time prolonged to 3w, the protein expression of PPARγ increased significantly (P < 0.05); the mRNA expression levels of BMP-2 in the high glucose group was lower than in the control group (all P < 0.05). (2)At the same intervening time, the expression of PPARγ mRNA decreased while BMP-2 mRNA increased (all P < 0.05) with the increasing concentration of MF (from 0 μmol · L-1 to 100 μmol · L-1); when the intervention time prolonged from 1w to 3w, the mRNA expression levels of PPARγ increased in all the high glucose groups while the mRNA expression levels of BMP-2 decreased (all P < 0.05). (3)When interviewed for 3w, the protein expression of PPARγ decreased gradually with the increasing concentration of MF in high glucose condition (P < 0.05). CONCLUSION: Pathological concentration of glucose could lead to poorly differentiated of osteoblasts, which might association with the enhanced mRNA and protein expression levels of PPARγ; MF can protect osteoblasts by restraining the expression of PPARγ.

ObjectiveTo evaluate the safety and efficacy of urinary kallidinogenase for recombinant tissue-type plasminogen activator (rt-PA) intravenous thrombolytic treatment in patients with acute cerebral infartion MethodsA randomized control study was applied. All 44 patients with acute cerebral infartion were randomized 1:1 to the experimental group (22 cases) and the control group (22 cases). Patients were administrated rt-PA(0. 9 mg/kg)in control group, and patients were given urinary kallidinogenase by intravenous drip (0.15 PNAU/d, for 7 days) after rt-PA intravenous thrombolytic treatment (0.9 mg/kg)in experimental group. The main evaluation index was the incidence of symptomatic intraeerebral hemorrhage within 24 hours, and the secondary assessing items were NIHSS and BI. ResultsThere was 1 case (4.6%) with symptomatic intracerebral hemorrhage in the experimental group and 2 (9.1%) in the control group (X2 =0.00, P= 1.000),and reinfarction rate showed a decreasing tendency in experimental group (18.2% vs. 31.8%, X2=1.091,P=0.296). Compared with the control group, the NIHSS scores were significantly lower 1,21,90 days after thrombolytic therapy (t=2.119, 2.913, 2.187);P=0.041, 0.0 06, 0.042),and the BI scores were obviously higher at 90 days after thrombolytic therapy in experimental group(t= 2.39,P= 0.012). ConclusionsWithout increasing the risk of intracerebral hemorrhage, urinary kallidinogenase may improve the curative effect for rt-PA intravenous thrombolytic treatment in patients with acute cerebral infartion

[Objective] To determine the effects against human simplex virus type I(HSV-1) and adenovirus type Ⅲ(ADV-3) anti-viral in vitro and in vivo of eye drop prepared by mixed extract of Heartleaf houttuynia and Lonicera japonica.[Methods] By using different numbers of host cells with cytopathic effect(CPE) as the observation index,models of HSV-1 infected Vero cells and ADV-3 infected HeLa cells were established to determine the anti-viral effects in vitro of HLD with different dosages.Rabbit eye conjunctivitis model infected by HSV-1 was established to observe the anti-viral effects in vivo of HLD with different dosages,and the biopsy specimens of pathogenic eye conjunctive tissue were examined by routine pathological methods.In these experiments,a commercial acilovir eye drop(ACV) was used as the control.[Results] The minimum dosages of HLD to completely inhibit the CPEs of HSV-1 and ADV-3,the host cells were 32 mg/ml and 64 mg/ml respectively.After treatment with HLD,the inflammatory reactions of rabbit eye conjunctivitis caused by HSV-1 infection were obviously weak or disappeared.The curative rates of 1,3 and 6 g/ml HLD treating experimental rabbit eye conjunctivitis were 50.0%,83.3% and 100% respectively.Furthermore,the curative effect using 6 g/ml HLD has no significant difference compared to that using ACV drop.[Conclusion] HLD has a certain anti-viral effect in vitro and a stronger anti-viral effect in vivo,indicating the potential of HLD for developing a new drug to prevent or treat viral eye conjunctivitis.

Objective To generate a comD gene knock-out mutant of Streptococcus pneumoniae,and determine the correlation of comD gene and the bacterial resistance against β-lactam antibiotics and understand the effect of closantel down-regulating comD, comE and comC mRNA levels. Methods A suicide plasmid pEVP3comD was constructed for comD gene knock-out and a comD gene knock-out mutant (comD-)was generated through homologous recombination and insertion inactivation. PCR and immunofluorescence method were used to identify the comD- mutant and real-time fluorescence quantitative PCR was applied to detect the changes of comD, comE and comC mRNA levels before and after closantel treatment in comD-mutant and wild-type strain. Double agar dilution method was performed to determine the sensitivity of comD- mutant and wild-type strain to penicillin G and cefotaxime. Results The comD gene in genome DNA of the generated comD- mutant was inactivated by sequencing and immunofluorescence detection. 50 μ mol/L or 100 μmol/L closantel had a function to down-regulate the comD, comE and comC mRNA levels ( P ＜ 0. 05) whereas 25 μmol/L closantel did not. Both the MIC values of penicillin G and cefotaxime inhibiting comD- mutant were 32 μg/ml which was significantly higher than that of wild-type strain (0.06 μg/ml and 1 μg/ml). Conclusion In this study a comD gene knock-out mutant of S. pneumoniae was successfully generated. There is a close correlation between comD gene and β-lactam antibiotics resistance of S. pneumoniae. Closantel has a function to inhibit the competence formation of S. pneumoniae through down-regulating the transcription levels of comD, comE and comC genes.

Adult ; Audiometry, Pure-Tone ; Case-Control Studies ; Evoked Potentials, Auditory, Brain Stem ; Hearing Loss, Noise-Induced ; diagnosis ; physiopathology ; Humans ; Male ; Middle Aged ; Noise, Occupational

Objective To evaluate the changes in the expression of hypoxia-inducible factor 1 alpha (HIF-1α) in the proliferated intima of pulmonary arterioles in rats with pulmonary hypertension.Methods Thirty-two pathogen-free male Sprague-Dawley rats,aged 2 months,weighing 200-230 g,were used in the study.Thirty-two rats were randomly divided into 4 groups with 8 animals in each group using a random number table:shamoperation group (group S),left pneumonectomy group (group P),monocrotaline group (group M),and left pneumonectomy combined with monocrotaline group (group PM).The rats underwent left pneumonectomy in P and PM groups.In M and PM groups,monocrotaline 60 mg/kg was injected subcutaneously at 7 days after operation.On day 35 after operation,mean pulmonary artery pressure (mPAP) and right ventricle systolic pressure (RVSP)were measured via direct puncture of right ventricle outflow tract with gauge 24 iv catheter connected to pressure transducer.At the end of experiment,the hearts and right lobes of the lung were excised for measurement of right ventricle hypertrophy index (RVHI) and the thickness of the medial layer (tunica media) of pulmonary arterioles (MT).Vascular occlusion score (VOS) was performed.The expression of HIF-1α,α-smooth muscle actin (α-SMA),and proliferating cell nuclear antigen (PCNA) in the pulmonary arterioles was determined.Results Compared with S group,MT was significandy increased,and the expression of α-SMA was up-regulated in P group,mPAP,RVSP,RVHI and MT were increased in M group,mPAP,RVSP,RVHI,MT and VOS were increased in PM group,and the expression of HIF-1α,α-SMA,and PCNA was up-regulated in M and PM groups.Compared with P group,mPAP,RVSP,RVHI and MT were significantly increased,and the expression of HIF1α,α-SMA,and PCNA was up-regulated in M and PM groups.Compared with M group,RVSP,RVHI,MT and VOS were si gnificantly increased,and the expression of HIF-1α,α-SMA,and PCNA was up-regulated in PM group.Conclusion The mechanism of pulmonary arteriole intimal proliferation is related to up-regulated HIF-1α expression and promoted proliferation of vascular smooth muscle cells in rats with pulmonary hypertension.

Objective To construct a prokaryotic expression system for expressing the phosphatase-encoding gene phpP in StkP/PhpP signaling couple in Streptococcus pneumonia ( S.pneumoniae) strains, and to further understand the phosphatase activity of the recombinant protein rPhpP.Methods The entire phpP gene of S.pneumoniae strain ATCC6306 was amplified by PCR.The PCR products were sequenced.A prokaryotic ex-pression system for expressing the phpP gene was constructed by the genetic engineering technique.The ex-pressed protein rPhpP and the solubility of rPhpP were assessed by SDS-PAGE and gel image analyzer.Ni-NTA affinity chromatography was performed to purify rPhpP.The changes of phpP gene transcription after the treat-ment with sublethal dosages of penicillin and cefotaxime were determined by real-time fluorescent quantitative RT-PCR.The functional domain in the sequence of the phpP gene and its type was analyzed by bioinformatic softwares.The activity of rPhpP in hydrolyzing the substrate of PP2C phosphotase was measured with Serine/Threonine Phosphotase Assay Kit.The enzyme kinetic parameters of rPhpP were calculated.Results The se-quence of the cloned phpP gene was identical with that reported in GenBank.The rPhpP in soluble form was ex-pressed in the constructed prokaryotic expression system.An increased expression of phpP gene at mRNA level was induced by sublethal dosage of penicillin or cefotaxime.The domain of PP2Cc type phosphatase was detec-ted in the sequence of phpP gene.The purified rPhpP protein could hydrolyze phosphopeptides [ RRA ( pT) VA], a substrate of PP2C type phosphatase, in a dose-dependent manner with Km and Kcat values of 277.35μmol/L and 0.71 S-1 ,respectively.Conclusion The protein encoded by phpP gene of S.pneumoniae was a PP2C type phosphatase.The expression of phpP gene could be enhanced by sublethal dosage of penicillin or ce-fotaxime.

Objective To study the changes of gastrin 17 (G17) and pepsinogen (PG) after gastric bypass surgery in gastric antrum resection, and the influences of different surgical methods on postoperative peptic ulcer. Methods Clinical data of 63 patients with gastric bypass surgery in our hospital from October 2013 to October 2015 were divided into resection of pyloric antrum group (n=33) and preserved pyloric antrum group (n=30). The values of G17, PGⅠ, PGⅡand PGⅠ/PGⅡwere detected by enzyme linked immunosorbent assay at 1 month, 6 months and 12 months after operation. The correlation between the different surgical methods and the incidence of peptic ulcer was analyzed between two groups. Results The G17 levels were significantly decreased in resection of pyloric antrum group 6 and 12 months after operation than those in preserved pyloric antrum group (P<0.05). Compared with preserved pyloric antrum group,PGⅠ and PGⅡ levels was significantly decreased 12 months after operation (P<0.05). There was no significant difference in the ratio PGⅠ/PGⅡat 1 month, 6 months and 12 months after operation between two groups (P>0.05). There was no significant difference in postoperative peptic ulcer between two groups (P>0.05). Conclusion Gastric bypass after resection of the pyloric antrum can reduce the postoperative secretion of G17, PGⅠ and PGⅡ, but which can not reduce the incidence of postoperative anastomotic ulcer.

Objective To construct a mutant strain of Streptococcus pneumoniae ( S.pneumoniae) with ciaH gene-knockout (ΔciaH) and to analyze the correlation between the ciaH gene and the bacterial re-sistance against β-lactam antibiotics.Methods The ciaH gene segament of S.pneumoniae strain ATCC6306 was amplified by PCR.The PCR product was sequenced after T-A cloning.A suicide plasmid pEVP3ciaH was constructed for the deletion of ciaH gene and then transformed into the ATCC 6306 strain by using the CaCl2 method .The mutant strain of S.pneumonia strain ATCC6306 with ciaH gene-knockout (ΔciaH) was genera-ted through homologous recombination , insertion inactivation and amphemycin screening , which was further identified by PCR , sequencing analysis and laser confocal microscopy .Double agar dilution method was used to detect the minimal inhibitory concentrations ( MICs ) of penicillin G ( PCN ) and cefotaxime ( CTX ) against theΔciaH mutant strain and the wild type strain .The differences between the MICs were further ana-lyzed.The changes of ciaH gene expression at mRNA level after treatment with 1/4 MIC of PCN or CTX were detected by real-time fluorescent quantitative RT-PCR ( qRT-PCR ) .Results The ciaH gene in the genomic DNA of the generated ΔciaH mutant strain was inactivated by insertion as indicated by PCR and se-quencing analysis .Results of the immunofluorescence assay showed that the ΔciaH mutant strain did not ex-press the CiaH protein .The MICs of PCN and CTX against the ΔciaH mutant strain were 32 μg/ml and 64μg/ml, respectively, which were significantly higher than that of the wild type strain (0.06 μg/ml and 1μg/ml) (P<0.01).The expression of ciaH gene at mRNA level was significantly elevated after treatment S.pneumoniae ATCC6306 strain with 1/4 MIC PCN or CTX (P<0.01).Conclusion The CiaH protein in the CiaH/CiaR two-component signaling system is involved in the resistance of S.pneumoniae against β-lac-tam antibiotics.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Liver ; pathology ; Liver Cirrhosis ; pathology ; Paeonia ; Rats ; Rats, Sprague-Dawley