Objective To study the inhibition of anti-PDGF on proliferation of cultured human retinal pigment epithelial cells(hRPE) in vitro.Methods hRPE were cultivated and were exposed to different concentrations of anti-PDGF(0,1?10-6,5?10-6,1?10-5,5?10-5 and 1?10-4 mg?L-1) respectively .Growth curves were measured with cell counting and the vitalities of cells were examined by percentage of vital cells and total cells.Using MTT staining colorimetric to measure the inhibitory rate.The changes of cell cycle of hRPE were collected and their growth were detected with FCM analysis and the morphological changes of cells were observed by light microscope and electron microscope.Results Anti-PDGF of 1?10-6mg?L-1 stimulated hRPE proliferation slightly.AntiPDGF at dosages ranging from 5?10-6mg?L-1 to 1?10-4mg?L-1 inhibited cell proliferation effectively in a dose-dependent and time-dependent manner(P

Objectives To explore the calcium signaling mechanism of STIM1 in breast cancer cells. Meth-ods After SiRNA interruption, Western blot and Transwell were used to measure protein expression of STIM1 and cell migration in MDA-MB-231 cells respectively. The relationship between STIM1 and SOCE calcium signaling were analysed by Laser confocal microscopy. Western blots were used to measure protein expression of FAK after si-lence STIM1. Results The numbers of cells without STIM1 were significantly lower than those cells with STIM1 by Transwell assay. STIM1 mediated SOCE in MDA-MB-231. Blocking SOCE might inhibite cells migration. Si-lence STIM1 did not affect the expression or activation of FAK in MDA-MB-231 cells. Conclusion STIM1 influ-ences cell migration through SOCE pathway in breast cancer cells, which is independent on the expression or activa-tion of FAK.

According to the sequence of P450 cDNA of Eleutherococcus senticosus, specific primers were designed. Frokaryotic ex pression vector pET30a-P450 was constructed and the prokaryotic expression conditions were optimized. Results showed that the BL21 after being transformed with the recombinant expression vector accumulated the high amount of recombinant protein. SDS-PAGE analysis showed that the recombinant protein was about 53 kDa. The recombinant accumulated the highest amount of recombinant protein af ter IPTG (1 mmol x L(-1)) at 27-37 degrees C for 24 h. Consequently P450 gene of E. senticosus could be expressed successfully by prokaryotic expression vector pET30a-P450. Induction temperature, IPTG concentration, medium type and amount of induction time could all influence the expression of target protein, but the impact strength was different.
Cloning, Molecular ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Eleutherococcus ; enzymology ; genetics ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Plant Proteins ; genetics ; metabolism ; Plasmids ; genetics ; metabolism ; Recombinant Proteins ; genetics ; metabolism

OBJECTIVETo explore the expression of Ghrelin and high mobility group protein B1 (HMGB1) in the serum and the intestinal tissue of sepsis model rats, and to evaluate the effect of electro-acupuncture (EA) at Zusanli (ST36) on the expression of HMGB1 and Ghrelin.

METHODSForty-eight male Wistar rats were randomly divided into four groups, i.e., the sham-operation (sham), the cecal ligation and puncture group (CLP), the CLP + EA at Zusanli (ST36) group (EA), and the CLP + Ghrelin receptor blocking agent + EA group (GHSRA), 12 in each group. A sepsis rat model was prepared by CLP. The incision of the abdominal wall was immediately sutured along the ventral midline for rats in the Sham group. In the EA group EA at Zusanli (ST36) was performed 20 min after CLP surgery with the constant voltage (2 - 100 Hz, 2 mA) for 30 min. In the GHSRA group, Ghrelin receptor blocking agent, [D-Arg1, D-Phe5, D-Trp79, Leu11]-substance P (700 nmol/kg), was administered through intravenous injection immediately after CLP, and 20 min later, EA at Zusanli (ST36) was performed in the same way as for rats in the EA group. Blood samples were withdrawn 12 h after CLP. The serum levels of Ghrelin and HMGB1 were detected using ELISA. Ghrelin expressions and the number of Ghrelin immunopositive cell in the jejunum were determined by immunohistochemistry. HMGB1 contents of the jejunum tissue were detected by Western blotting.

RESULTSCompared with the Sham group, the number of serum immunopositive cells and the expression of HMGB1 in the jejunum tissue significantly increased and levels of Ghrelin and the expression rate of immunopositive cells significantly decreased in the CLP group (P < 0.05). Compared with the CLP group, the number of serum immunopositive cells and the expression of HMGB1 in the jejunum tissue significantly decreased, but levels of Ghrelin and the expression rate of immunopositive cells significantly increased in the EA group (P < 0.05). Compared with the EA group, the number of serum immunopositive cells and the expression of HMGB1 in the jejunum tissue significantly increased in the GHSRA group (P < 0.05), but there was no statistical difference in levels of Ghrelin between the two groups (P > 0.05). The serum level of HMGB1 was negatively correlated with Ghrelin in the Sham group, the CLP group, and the EA group (r = -0. 528, P < 0.01).

CONCLUSIONSEA at Zusanli (ST36) could inhibit the expression of HMGB1 in the jejunum of septic rats, and promote the expression of Ghrelin. The expression of HMGB1 was inhibited by Ghrelin receptor blocking agent, which suggested that the anti-inflammation of EA at Zusanli (ST36) might be associated with Ghrelin.

Animals ; Disease Models, Animal ; Electroacupuncture ; Ghrelin ; metabolism ; HMGB1 Protein ; metabolism ; Jejunum ; metabolism ; Male ; Rats ; Rats, Wistar ; Sepsis ; metabolism

OBJECTIVETo observe the effect of Yifei Qinghua Granule (YQG) on vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), angiostatin, and endostatin in tumor tissue of Lewis Lung cancer mice, and to explore its anti-tumor mechanisms.

METHODSTotally 70 C57BL/6 mice were randomly divided into the model group, the low, medium, and high dose YQG groups, the gefitinib group, the gefitinib plus medium dose YQG group, and the cyclophosphamide (CTX) group, 10 in each group. The models were established by subcutaneously injecting Lewis lung cancer cells from the right axilla of C57BL/6 mice. Mice in the model group were given with 0.4 mL pure water by gastrogavage, once daily. Mice in the low and medium dose YHG groups were given with YHG at the daily dose of 5 and 10 g/kg by gastrogavage, once daily. Those in the high dose YHG group were given with YHG at 10 g/kg by gastrogavage, twice daily. Those in the gefitinib group were given with gefitinib 100 mg/ kg by gastrogavage, once daily. Those in the gefitinib plus medium dose YHG group were given with gefitinib at 100 mg/kg by gastrogavage in the morning and YHG at 10 g/kg by gastrogavage in the afternoon. All medication was started from the 2nd day of inoculation, lasting 14 successive days. Those in the CTX group were given CTX at 60 mg/kg by peritoneal injection on the 3rd and the 7th day of the experiment. Mice were sacrificed at the fifteenth day of the experiment. Tumors were taken out. Expressions of VEGF, bFGF, angiostatin, and endostatin in the tumor tissue were detected using immunohistochemical assay.

RESULTSCompared with the model group, the expression of VEGF significantly decreased, expressions of angiostatin and endostatin significantly increased in each group (P < 0.01). The expression of bFGF significantly decreased in the gefitinib group (P < 0.05). There was no statistical difference in VEGF among all groups (P > 0.05). The angiostatin expression was significantly higher in the CTX group than in the low dose YQG group (P < 0.01). The expression of endostatin was significantly higher in the high dose YQG group and the gefitinib plus medium dose YQG group than in the low and the medium dose YQG groups (P < 0.01). The expression of endostatin was significantly higher in the gefitinib plus medium dose YQG group than in the gefitinib group (P < 0.05).

CONCLUSIONThe action mechanism of YQG in treating lung cancer might be achieved through reducing the expression of angiogenesis promoting factor VEGF and increasing expressions of angiogenesis inhibitors angiostatin and endostatin.

Angiostatins ; metabolism ; Animals ; Carcinoma, Lewis Lung ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Endostatins ; metabolism ; Fibroblast Growth Factor 2 ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Phytotherapy ; Vascular Endothelial Growth Factor A ; metabolism

Objective Few reports are seen about the effects of cooling blood and removing stasis on nephrin and podocin . This study was to evaluate the therapy of cooling blood and removing stasis for Henoch -Sch?nlein purpura nephritis ( HSPN) and its ac-tion mechanisms by observing the effects of Danshao Granular Ⅲ on hematuresis , proteinuria and the expressions of nephridial nephrin and podocin in HSPN rats . Methods Twenty-one SD male rats were e-qually randomized to a blank control , an HSPN model , and a Dan-shao group.At 13 weeks after modeling , the animals in the model group were treated intragastrically with distilled water , while those in the Danshao group with Danshao Granular Ⅲtwice daily for 4 weeks. Then, the urinary red blood cell ( RBC) count was examined , the
24 h urinary protein quantity determined , the glomerular mesangial changes observed under the light microscope , the protein expres-sions and distributions of nephrin and podocin detected by indirect immunofluorescence , and their mRNA expressions determined by re-al-time PCR. Results The urinary RBC count and 24 h urine protein quantity were significantly higher in the HSPN model than in the blank control group (26.5/HP vs 0.3/HP and [2.214 ±1.090]g/24 h vs [0.624 ±0.354]g/24 h, both P<0.01).The model rats showed obvious pathological changes in the renal tissue .The urinary RBC count and 24 h urine protein volume were remarkably de-creased in the Danshao group as compared with the models (0.8/HP vs 26.5/HP and [1.000 ±0.651]g/24 h vs [2.214 ±1.090] g/24 h, both P<0.01).The pathological changes in the renal tissue of the Danshao group were reduced in comparison with those of the model group.The protein expression of nephrin was higher in the former than in the latter (65.975 ±14.414 vs 43.520 ±0.632, P<0.01) and so was that of podocin though with no statistically significant difference (P>0.05).The distributions of nephrin and podocin were improved after Danshao treatment .The mRNA expressions of nephrin and podocin were markedly higher in the Danshao group than in the HSPN models (0.530 ±0.089 vs 0.117 ±0.021 and 0.490 ±0.160 vs 0.033 ±0.025, P<0.05). Conclusion Danshao Granular Ⅲ, with its main action mechanisms of cooling blood and removing stasis , can effectively reduce urinary RBC count and urinary protein quantity and improve the symptoms of HSPN in rats .

Objective To explore changes of gray matter volume in patients with obsessivecompulsive disorder (OCD) in Chinese Han population using optimized voxel-based morphometry (VBM) ,and investigate its relationship with clinical symptoms. Methods Twenty patients with OCD and 20 age,sex and handedness matched healthy controls were scanned using 3D-T1 images on a 3.0 T MR system. The high resolution T1WI was preprocessed according to the optimized VBM protocol in Statistical Parametric Mapping (SPM5). Two-sample t test was performed to characterize the differences of the gray matter volume (GMV) between the OCD patients and healthy controls, and the correlations between the GMV and symptom severity and cumulative illness duration were examined using Pearson correlation in SPSS 16. 0, respectively.Results Compared to controls, OCD patients demonstrated increased GMV in left thalamus, right thalamus and left cerebellum after false discovery rate (FDR) correction. No areas of significantly decreased GMV was observed in OCD patients in relative to healthy controls. The mean eigenvalue ranged from 0. 5782 to 0. 889 representing the left thalamus volume of OCD patients was 0. 6813 ± 0. 0718, and that ranged from 0. 5546 to 0. 9062 was 0. 6869 ± 0. 0808 tor right thalamus. The mean eigenvalues were positively correlated in bilateral thalamus (r = 0. 94, P ＜ 0. 01). Conclusion Using optimized VBM, the current research indicates that the pathophysiology of OCD is associated with GMV abnormalities not only in corticostriato-thalamo-cortical (CSTC) circuit, but also in the cerebellum.

OBJECTIVETo systematically review the effectiveness of bone substitute augmentation combined with internal fixation versus internal fixation alone in treating proximal femoral fractures in the elderly.

METHODSSubject term and keywords were searched from Pubmed, Cochrane databases and CNKI from database foundation to August 2015. Randomized controlled studies and qusi-randomized controlled studies on bone substitutes augmentation combined with internal fixation versus internal fixation alone for the treatment of proximal femoral fractures in the elderly were chosen. Postoperative re-displacement, re-operation rate, complications (infection and bone ununion), functional outcome, quality of life scores and muscle strength were seen as outcome indicators. Enumeration data were statistical analyzed by risk difference and 95% confidence interval. Measurement data were analyzed by standardized mean difference and 95% confidence interval. If the same measurement data were evaluated by different standards in different studies, standardized mean differences and 95% confidence interval were used. The methods of statistical analysis were used by Cochrane databases.

RESULTSEleven RCTs (677 patients) were included. Meta-analysis results indicated that bone substitutes augmentation combined with internal fixation occurred fewer re-displacement [SMD = -0.75, 95% CI (-1.03, -0.47)] and obtained better function [SMD = 0.40, 95% CI (0.20, 0.59)]. While there were no significant differences in reoperation rate [RD = 0.02, 95% CI (-0.05, -0.09)], pain at 1 week after operation [MD = -1.79, 95% CI (-13.55, -9.96)], pain ranged from 6 to 8 weeks [MD = -7.24, 95% CI (-20.07, -5.59)], postoperative pain at 12 weeks [MD = -0.32, 95% CI (-4.9, -3.55)], muscle strength [MD = 1.25, 95% CI (-6.98, -9.48)], bone ununion [RD = 0.02, 95% CI (-0.01, -0.05)] and postoperative complications [MD = 0.01, 95% CI (-0.03, -0.04)].

CONCLUSIONCompared with single internal fixation, bone substitutes augmentation combined with internal fixation for the treatment of proximal femoral fractures in the elderly less occur re-displacement and could obtain better functional recovery.

Aged ; Aged, 80 and over ; Bone Substitutes ; administration & dosage ; Female ; Femoral Fractures ; surgery ; therapy ; Fracture Fixation, Internal ; Humans ; Male ; Middle Aged ; Randomized Controlled Trials as Topic ; Treatment Outcome

OBJECTIVETo investigate clinical effects of strategy by stages for preventing respiratory complications of patients with acute cervical spinal cord injury (ACSCI).

METHODSFrom September 2009 to May 2013,the clinical data of 91 patients with ACSCI underwent surgery were retrospectively analyzed. Among the patients, 42 patients were divided into in-stages group, including 30 males and 12 females with an average age of 50 years old (ranged 28 to 76) which were treated with strategy by stages for preventing respiratory complications; others 49 patients which were not treated with the strategy regarded as control group, including 38 males and 11 females with an average age of 47 years old (ranged 30 to 77). All of them had definite history of trauma, and were admitted to orthopaedics within 48 h after trauma. In in-stages group, respiratory muscle strength training, high-dose ambroxol using and other treatment were performed to prevent respiratory complilcations according to preoperative, intraoperative and postoperative stage. While in control group, there were no systematic and effective measures utilized. Chi-square test was used to evaluate the difference for respiratory complications rate, the rate of tracheostomy or intubation and mortality caused by the respiratory complications between two groups.

RESULTSTen patients developed with respiratory complications in in-stages group (7 patients with pneumonia, 1 with atelectasis and 2 with respiratory failure), among which 3 patients underwent tracheostomy or intubation. In control group, 24 patients developed with respiratory complilcations (15 with pneumonia,3 with atelectasis and 6 with respiratory failure), among which 11 patients underwent tracheostomy or intubation. There was significant difference between two groups (χ2 = 6.12, 4.07; P = 0.013, 0.044). Five patients died because of respiratory complications, one case were in in-stages group and 4 in control group. There was significant difference between two groups (χ2 = l.39, P = 0.238).

CONCLUSIONThe strategy by stages is an effective method for preventing respiratory complications of ACSCI and can reduce the respiratory complications rate and improve the prognosis of respiratory complications.

Acute Disease ; Adult ; Aged ; Cervical Cord ; injuries ; Female ; Humans ; Male ; Middle Aged ; Pneumonia ; prevention & control ; Pulmonary Atelectasis ; prevention & control ; Respiratory Insufficiency ; prevention & control ; Retrospective Studies ; Spinal Cord Injuries ; complications ; Tracheostomy

Objective To observe enhancing effect of nerve regeneration on peripheral nerve defect models bridged by VPA/PRGD conduit combined with adipose derived mesenchymal stem cells (ADSCs).Methods From February,2013 to August,2014,the hollow nerve scaffolds were constructed by PRGD and VPA.The activity of AD SCs proliferation was tested through the method of CCK-8.Moreover,the effect of VPA/PRGD conduit combined with ADSCs on peripheral nerve regeneration was evaluated,as well as recovery of motor function following sciatic nerve resection in rats.A 10 mm sciatic nerve deficit was created in a rat model and bridged by VPA/PRGD conduit combined with ADSCs (group A),VPAfPRGD conduit (group B) and autograft (group C) respectively.The results was analyzed by the method of group t-test of SPSS 19.0.Results At 3 d,6 d,9 d,CCK-8 test showed that the OD value of groups VPA/PRGD and control has no significance(P ＞ 0.05).At 12 weeks after surgery,the numbers of regenerated nerve proximal to the tube of group A (268±7.48),group B (269±6.86) and group C (271±7.55),had no significant difference between two groups(P ＞ 0.05).The numbers of regenerated nerve in the tube of group A (257±6.19) and group C (260±5.60) were significantly higher than those of group B (229±5.08) (P ＜ 0.05).There was no significant between groups A and C (P ＞ 0.05).The numbers of regenerated nerve distal to the tube of group A (246±5.89) and group C (247±5.02) were significantly higher than those of group B (214±7.55) (P ＜ 0.05).There was no significant between groups A and C (P ＞ 0.05).Conclusion These promising results illustrate that this novel VPA/PRGD combined with ADSCs conduit can obviously facilitate the regeneration of injured nerve in rats.