Objective To investigate the effects of the three methods of decocting with deslag, decocting without deslag, and double decocting on the content of nine ingredients baicalin, baicalein, ginsenoside Re, ginsenoside Rb1, monoammonium glycyrrhizinate hydrate, liquiritin, 6-gingerol, berberine hydrochloride, palmatine hydrochloride, and total flavonoids in Banxia Xiexin Decoction (BXD). Methods Nine index components were determined by HPLC. The HPLC analysis was performed on Welch Ultimate XB-C18 column (250 mm × 4.6 mm, 5 μm) with mobile phase of acetonitrile-0.1% phosphate aqueous solution for gradient elution; And carried out at column temperature of 28 ℃, volume flow of 0.9 mL/min, and detection wavelength of 203, 252, 280, and 355 nm. The total flavonoids were determined by colorimetry. Results Nine kinds of ingredients and total flavonoids could be detected in three different decoctions. In the method of decocting with deslag, baicalin, baicalein, ginsenoside Rb1, monoammonium glycyrrhizinate hydrate, and liquiritin increased by 10.01%, 12.88%, 29.09%, 16.75%, and 15.02%, respectively, compared with decocting without deslag; It decreased by 5.54%, 4.15%, 14.49%, 7.85%, and 9.18%, respectively compared with double decocting; Ginsenoside Re, 6-gingerol, berberine hydrochloride, and palmatine hydrochloride increased by 37.90%, 3.78%, 5.33%, and 5.99% compared with decocting without deslag, respectively; compared to the double decocting methods, it increased by 1.07%, 11.57%, 3.41%, and 1.93%. The total flavonoids increased 22.61% higher than decocting without deslag and 6.54% higher than double decocting. Conclusion: The results can effectively reflect the quality difference of different decocting methods. Among the three methods of decoction, the method of decocting without deslag has significantly improved the dissolution of the active ingredients of each component in the decoction, and improve the clinical efficacy of BXD to a certain extent. It provides a good experimental basis for the decocting without deslag method used in Zhang Zhongjing’s Treatise on Febrile Diseases.

Objective: To clone hGLP-1 cDNA in the pBS SK(+/-)vector and construct the expression vector of pGEX-4T-3/hGLP-1cDNA to express GST-hGLP-1 fusion protein. Methods: The hGLP-1 cDNA was constructed by 6 synthetic oligonucleotides fragments, followed by the procedure of annealing and ligation with oligonucleotides fragments. The hGLP-1 cDNA was cloned into the pBS SK(+/-) vector, and was selected by α-complementation. It was confirmed by DNA sequening, then inserted into the MCS of the fusion expression vector pGEX-4T-3. The recombinant vector was transformed into E. coli TG1. Results: The recombinant plasmid DNA was digested with restrictive endonuclease BamHⅠand XhoⅠ. The result demonstrated that the hGLP-1 cDNA was successfully inserted into the pGEX-4T-3 vector and fusion protein GST-hGLP-1 had been expressed in SDS-PAGE. Conclusion： Expression of GST-hGLP-1 fusion protein can provide foundation for obtaining a larger quantity of recombinant hGLP-1 for experimental and clinic studies.

Objective: To clone hGLP-1 cDNA in the pBS SK(+/-)vector and construct the expression vector of pGEX-4T-3/hGLP-1cDNA to express GST-hGLP-1 fusion protein. Methods: The hGLP-1 cDNA was constructed by 6 synthetic oligonucleotides fragments, followed by the procedure of annealing and ligation with oligonucleotides fragments. The hGLP-1 cDNA was cloned into the pBS SK(+/-) vector, and was selected by α-complementation. It was confirmed by DNA sequening, then inserted into the MCS of the fusion expression vector pGEX-4T-3. The recombinant vector was transformed into E. coli TG1. Results: The recombinant plasmid DNA was digested with restrictive endonuclease BamHⅠand XhoⅠ. The result demonstrated that the hGLP-1 cDNA was successfully inserted into the pGEX-4T-3 vector and fusion protein GST-hGLP-1 had been expressed in SDS-PAGE. Conclusion： Expression of GST-hGLP-1 fusion protein can provide foundation for obtaining a larger quantity of recombinant hGLP-1 for experimental and clinic studies.

PurposeTo investigate the clinical value of inverted X-ray plain film and MRI examination in the diagnosis of congenital anorectal malformation (CARM). Materials and Methods Thirty-eight cases with operatively proved anorectal malformation were reviewed; inverted X-ray plain film and MRI examination were performed in all patients before surgery. The relationship between the rectum blind side and pubococcygeal line (PC line), and the type of anal atresia was determined, to compare the diagnostic accuracy of inverted plain film with MRI for CARM typing. Results Of all the 38 cases, 19 cases were with low imperforate anus, 8 cases with median imperforate anus, and 11 cases with high imperforate anus. The accuracy rate of inverted X-ray plain film and MRI examination for the diagnosis of CARM typing was 92.1% (35/38) and 97.4% (37/38) respectively, and the difference between them was not statistically significant (χ2=1.37, P>0.05). 7 cases of fistula, 5 cases of spinal cord malformations and 1 case of right kidney agenesis can be clearly demonstrated on MRI. Conclusion Both inverted X-ray plain film and MRI can diagnose the typing of CARM accurately, but MRI is also able to diagnose the fistula, visceral, spinal cord lesion and other abnormalities accompanied with CARM, while reducing the dose of X-ray radiation and damage in children, thus has higher clinical application value compared with inverted X-ray plain film.

OBJECTIVETo observe clinical effects of posterior short-segment fixation with undermining decompress by posterior ligament complex for the treatment of upper lumbar burst fractures.

METHODSFrom October 2010 to March 2013,23 patients with upper lumbar burst fractures (Denis B type) were treated by posterior short-segment fixation with undermining decompress by posterior ligament complex. There were 18 males and 5 females aged from 26 to 64 years old with an average of 45.7 years old. Twelve patients were caused by falling down, 5 cases were caused by traffic accident, 4 cases were the bruise injury caused by heavy object and 2 cases were caused by other injury. Fourteen patients were L1 fracture and 9 patients were L2 fracture. Thirteen patients were combined with nerve injuries (degree D according to ASIA classification). Internal fixation were removed from 12 to 20 months with an average of 14.3 months. JOA scores and imaging changes were recorded and compared at different time points.

RESULTSAll patients were followed up from 18 to 24 months with an average of 20.4 months. Thirteen patients with nerve injuries were completely recovered at 3 to 6 months after operation. JOA score at 1 year after operation was 20.63 ± 0.92, and 20.38 ± 1.06 at 3 months after removal of internal fixation,which were improved obviously than 9.90 ± 2.73 at 3 months after operation. (P > 0.05) Anterior height of injured vertebrae, vertebral body angle and local Cobb angle was (95.0 ± 0.53)%, (2.78 ± 1.36) and (2.43 ± 1.52) °respectively, and improved obviously than that of before operation (P < 0.05). There was no statistical significance in JOA scores at 3 months after removal of internal fixation and 1 year after operation (P > 0.05).

CONCLUSIONposterior short-segment fixation with undermining decompress by posterior ligament complex for the treatment of upper lumbar burst fractures has advantages of minimally invasive, could effective recover vertebrae height, maintain stability of spine, decrease low back pain. It is a safe and effective operative method.

Adult ; Decompression, Surgical ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Lumbar Vertebrae ; injuries ; Male ; Middle Aged ; Spinal Fractures ; surgery

Objective To evaluate the dynamic contrast-enhanced MRI (DCE-MRI)in detecting experimentally induced testicular ischemia. Methods Thirty healthy male New Zealand rabbits were randomly assigned into 6 groups. There were 5 rabbits in each of the following experimental groups: ( 1 ) Normal control, (2) Sham-operated, (3) ischemia of 3 h group, (4) ischemia of 6 h group, (5) ischemia of 12 h group, (6) ischemia of 24 h group. In all experiment groups, the right testis served as the internal control while the left testis served as the experimental side. DCE-MRI for each animal lasts about 10 minutes. Signal enhanced ratios (SERs) of ROI for both sides of each group were calculated by a computer, and parameters of SERs of 30 s, 75 s, 120 s and maximal SER were used for statistical analysis.Time intensity curves (TICs) were made for two sides of each group via Excel 2003 software and classified into 4 types. Statistical analysis was performed to compare the differences of SERs between left and right testis by two independent Kolmogorov-Smirnov test. Results In group I and 2, significant enhancement was observed on both testes of 10 rabbits. The enhancement decreased gradually with the elongation of ischemia in torsion groups. Three cases of type Ⅰ and 2 cases of type Ⅱ were observed in group 1,5 cases of type Ⅰ in group two, 2 cases of type Ⅰ and 3 cases of type Ⅱ b in group three, 2 cases of type Ⅰ and 2 cases of type Ⅱ b in group four, 5 cases of type Ⅱ b in group five and 5 cases of type Ⅲ in group six were noticed in the left testes. And in TICs of right testes, all cases showed TICs of type Ⅰ except 2 cases of type Ⅱ a in group six. In four torsion groups, the values for SER75 of the left side were 0. 084%, 0. 076%, 0.164% and 0.065%, while the right side were 0.255%, 0.410%, 0.586% and 0.302% (P ＜0.05). The values for SER120 in group three, five and six were 0.221% , 0.158% and 0.059% for the left side, and 0.405%,0.522% and 0.207% for the right side(P ＜0.05). The values for MSER in group three, five and six were 0.217% ,0.164% and 0.072% for the left side, and 0.405%, 0.586% and 0.302% for the right side(P ＜0.05). Conclusion DCE-MRI technique may be useful in the diagnosis of testicular torsion, which shows potential in the clinical application.

?AIM:To analyze the clinical features of Terrien marginal degeneration ( TMD) .?METHODS:Fifty-six patients(90 eyes) admitted in our hospital from January 2013 to January 2015 were selected as observation group.At the same period, 56 healthy (88 eyes) for corneal examination were as control group to analyze the clinical features of TMD. With immunohistochemistry and enzyme linked immunoassay method ( ELISA), the levels of HLA-DR, HLA-DQ and tumor necrosis factor-α( TNF-α) in blood samples of TMD patients were tested.?RESULTS:The transparent degree of the eye, lipid deposition in TMD patients with early, advanced, swelling and hole-wearing period were significantly different (χ2=10.85,χ2=65.32, P<0.05).Astigmatism in TMD patients with early and advanced, swelling and hole -wearing period were significantly different (P<0.05).The levels of HLA-DR, HLA-DQ and TNF-αin blood samples between the two groups were significantly different ( t=45.326, t=23.564, t=19.86, P<0.05).?CONCLUSION: Terrien's marginal degenerative is an inflammatory disease characterized by increased levels of TNF-α, HLA-DQ, and HLA-DR in peripheral blood, decreased corneal transparency, astigmatism and lipid deposition.This research provides experiment evidence for the mechanism of TMD.

Objective To investigate whether the effect of puerarin on right ventricle hypertrophy of pulmonary hypertensive rats induced by chronic hypoxia-hypercapnia was related to new peptide Apelin or its receptor(APJ).Methods Thirty male Sprague-Dawley rats were randomly divided into three groups, they are control group,hypoxia-hypercapnia 4-week model group,and hypoxia-hypercapnia 4-week plus puerarin group.The concentrations of Apelin-36 protein in plasma and homogenate of right ventricular muscle were detected by radioimmunoassay.The mRNA expressions of Apelin and APJ in right ventricu- lar muscle were measured by semi-quantitive reverse transcription polymerase chain reaction(RT-PCR). Results The weight ratio of right ventricle to left ventricle plus septum[RV/(LV+S)] in model group was significantly higher than that in control group(P0.05).The plasma concen- tration of Apelin-36 protein in model group was significantly higher than that in control group(P

AIMTo investigate the effects of L-arginine liposome on nitric oxide(NO) and nitric oxide synthase gene (NOS mRNA) in rats chronically exposed to hypoxia-hypercapnia.

METHODSFourty male SD rats were randomly divided into four groups (n=10): normal control group(NC), hypoxia-hypercapnia group (HH), hypoxia-hypercapnia + L-arginine group(HL) and hypoxia-hypercapnia + L-arginine liposome group (HP). Contents of NO in plasma were measured using colorimetric analysis. Expression of nitric oxide synthase gene were measured with situ hybridization.

RESULTS(1) The mean pulmonary artery pressure(mPAP) and weight ratio of right ventricle to left ventricle and septum(RV/LV + S) of HP group were obviously lower than those of HH group and HL group. (2) The NO contents in plasma of HP group were obviously higher than those of HH group and HL group (P < 0.01). (3) Situ hybridization showed the average value of integral light density(LD) of ecNOS mRNA in pulmonary arterioles was significantly higher in rats of HP group than that of HH group and HL group (P < 0.01). (4) Light microscopy showed WA/TA (vessel wall area/total area) and PAMT (media thickness ratio of pulmonary arterioles) were significantly lower in rats of HP group than those of HH group (P < 0.01).

CONCLUSIONL-arginine liposome could lower the mPAP and lighten the remodeling of pulmonary arterioles of the rats chronically exposed to hypoxia-hypercapnia than L-arginine does. It suggests that L-arginine liposome significantly promotes the membrane transportin of L-arginine.

Animals ; Arginine ; pharmacology ; Gene Expression ; Hypercapnia ; metabolism ; Hypoxia ; metabolism ; Liposomes ; pharmacology ; Male ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Rats ; Rats, Sprague-Dawley

The objective of this study was to investigate the effect of cytomegalovirus (CMV) infection on actin and microfilament in human embryo fibroblast cells (HF) and to explore the possible relationship with CMV replication. The cell shape was observed by microscopy after the infection of CMV, RT-PCR assay was used to detect the mRNA expression of beta-actin gene, while Westen-blot was used to measure the level of beta-actin protein. CMV immediately early antigen (IE) in HF cells was analyzed by indirect immunofluorescence assay. Microfilament alteration was determined by cytoskeleton fluorescence probe. The results showed that CMV IE was observed in more than 95% of HF cells after infection, primarily located in nucleus. HF cells infected by CMV changed from thin shuttle shape to round and thick ball shape, even detached from wall. Beta-actin got a significant and gradual decreasing of mRNA level in time-dependent manner (P < 0.05). Compared with uninfected group, the expression of beta-actin protein decreased to (74.2 +/- 13.4)% at 96 hours after infection (P < 0.05). In infected HF cells, microfilaments were ruptured, arranged turbulently, as well as cells merged and fluorescence density of microfilament obviously reduced. It is concluded that cytomegalovirus can induce alteration of actin and microfilament, which may be helpful for CMV to infect, replicate and reactivate in host cells.
Actin Cytoskeleton ; metabolism ; ultrastructure ; Actins ; metabolism ; Cell Line ; Cytomegalovirus Infections ; metabolism ; pathology ; Fibroblasts ; pathology ; ultrastructure ; virology ; Humans