Objective To construct the ?-gal reporter genes containing the 5′-end flanking of endothelial-overexpressed lipopolysaccharide-associated factor 1 (EOLA1) gene in different sequence lengths and identify the sequence, which regulates the gene expression of EOLA1 by the ?-gal analysis system. Methods The target sequences were amplified by the method of genome walker, and were inserted into the upstream of ?-gal gene located in the ?-gal enhancer vector by the directional clone technique respectively; the regulative sequence was identified by analyzing the ?-gal activities of reconstructed plasmid in ECV304 cells. Results The regions, containing 2 659 bp and 1 951 bp upstreaming from exon 1, significantly stimulated the reporter gene activity as compared with that of the ?-gal control vector in transfected cells. But the region, containing 361 bp upstreaming from exon 1, did not stimulate the reporter gene activity. Conclusion There is an up-regulative element of gene transcription in the region of -361 to -1 951 bp in EOLA1 gene upstream.

Objective To investigate the relationship between plasma triglycerides ( TG ) , cholesterol (TC)levels and breast cancer.Methods 105 cases of breast cancer(age from 22 to 69 years)and 261 cases of healthy control(age from 16 to 52 years)admitted from May.2014 to Oct.2014 were studied.TG and TC were tested.Subgroup analysis was performed by age (<35, 35-45, ≥45 years) and BMI( <18.5, 18.5-24, ≥24).Results TG level in breast cancer group was significantly higher than that of the healthy control group (t=-3.97,P=0.00), and TC level was lower than that in the healthy control group (t=2.49, P=0.01).TG lev-el of breast cancer patients≥45 years old was higher than that in the healthy control group ( t=-4.31, P=0.00).TC level of breast cancer patients <35 years and from 35 to 45 years was lower than that of the healthy control group(t=3.12, P=0.00及t=2.41, P=0.02); TG levels in breast cancer group were significantly lower than those of the healthy control group for all BMI subgroup ( t=-3.67, P=0.01 for BMI<18.5; t=-3.87, P=0.00 for BMI from 18.5 to 24.0;t=-2.01, P=0.04 for BMI≥24).TC level of breast cancer patients was higher than that of the healthy control group for patients whose BMI <18.5(t=-3.41, P=0.01) and it was lower than that of the healthy control group for patients whose BMI≥24.Conclusions The relation-ship between TG , TC and breast cancer is associated with age .Compared with healthy group , breast cancer pa-tients≥45 years have a higher TG level and breast cancer patients <45 years have a lower TC level .

Objective To analyze the relationship between the number of pelvic ring fractures, the location of fracture and Tile type of pelvic fracture and pelvic fracture healing status. To discuss how to understand the pelvic malunion, severe pelvic malunion and bony pelvis deformity （destruction） in Classification of the Impairment Related to Injury. Methods One hundred and ninety-eight cases of evaluation of body impairment class of pelvic fracture caused by traffic accident accepted by Zhongcheng Institute of Forensic Science in 2016 were collected. The cases were statistically analyzed in terms of the distribution of the number of pelvic ring fracture, the location of pelvic ring fracture and Tile type of pelvic fracture in various healing statuses. Results There were 16 cases without pelvic ring fracture, 18 cases of single pelvic ring fracture, 91 cases of 2 pelvic ring fractures, 73 cases of 3 or more pelvic ring fractures, 136 cases of anterior or posterior pelvic ring fractures, 46 cases of simultaneous anterior and posterior ring fractures. There were 34 cases of Tile A type, 130 cases of Tile B type and 24 cases of Tile C type. Statistical analysis results showed that, the differences in distribution of the number of pelvic ring fracture, the location of pelvic ring fracture and Tile type of pelvic ring fracture in various healing statuses had statistical significance （P<0.05）. Conclusion In determination of the healing status of pelvic fracture, preliminary judgment can be made by using the number of pelvic ring fracture, status of pelvic ring fracture and Tile type of pelvic fracture. Caution is required when dealing with single pelvic fractures （including dislocations）, and determining the destruction of bony pelvis.
Fracture Healing ; Fractures, Bone ; Humans ; Joint Dislocations ; Pelvic Bones ; Pelvis

Objective To investigate the characteristics of the road traffic accident (RTA) casualties in Shanghai in 2003.Methods.The characteristics of RTA casualties were analyzed retrospectively on the data of Public Security Bureau and other related information.Results There were 54 197 RTA cases in 2003 with 1 406 death and 11 178 injured.The incidence rate and fatality of RTA was 311.48/10~4 and 11.17% respectively.The vehicle or population injury rate and mortality were 64.24/10~4,8.08/10~4,83.31/10~5 and 10.48/10~5,respectively.The fatality of suburb was higher than that of urban.Most of the casualties were bicyclist,pedestrians and motorcyclist,ranged mainly from 21 to 50 years old. Head injury and multiple traumas were the leading cause of death.Conclusion The RTA related injury had become a public health problem.The key strategies to decrease the fatality and disability of RTA were to propagandize the traffic safety perception in citizen and enforce the construction of rescue system for RTA including the EMSS.

ObjectiveTo observe the effect on repairing facial nerve injury of rabbits by neural stem cells and autologous fasia. Methods22 rabbits with transected facial nerve were divided into 2 groups randomly, control group (8 rabbits,15 sides totally), which transected facial nerve were wrapped by autologous fasia, and treament group (14 rabbits, 20 sides totally), which were wrapped by neural stem cells and autologous fasia. Six weeks after transplantation, neuro-electrophysiological test, immunohistochemical examination were done. The number and thickness of myelin in the re-connected area of transected facial nerve were observed. ResultsThe transplanted animals recovered much better than that in control group (P<0.05). Immunohistochemical examination showed a great deal of BrdU positive cells around the re-connected area of transected facial nerve. Immunohistochemical staining also found plenty of regenerative myelins in this area in the treatment group. While in control group, there were no BrdU positive cells and only a few of regenerative myelins in the same area. ConclusionTransplantation of neural stem cells combined with autologous fasia might become the new method to treat facial nerve injury.

ObjectiveTo explore the method of co-culture of Schwann cell(SCs) with fascia and provide experimental basis for repairing transected nerve.MethodsSCs were co-cultured with fascia.Double staining by anti-BrdU and anti-S-100,S-100 fluorescent staining and anti-BrdU staining were used.ResultsThere were a plenty of SCs around fascia proliferated rapidly and disposed in parallel. SCs could be distinguished from fibroblastic cells by S-100 fluorescent staining and also be staining positive by anti-BrdU antibody,implying their high proliferous ability. Anti-BrdU and anti-S-100 staining showed numerous double staining positive SCs on the fascia: nucleus was stained deep blue while cytoplasm was stained red.ConclusionMany SCs with high proliferous ability were seen on the fascia, which can be used to repair transected nerve.

Objective To study the anti-inflammatory,antitussive,and analgesia effects of Jinhua qingre capsules.Methods The anti-inflammatory effect was assessed by the methods include xylene-induced mouse auricular swelling and histamine-induced pigment oozing from skin vessel in rats;The antitussive and analgesic effect were assessed by ammonia water induced cough model and acetic acid-induced twisting method.Results In anti-inflammation experiment,the high dose (12 g·kg-1) and moderate (6 g·kg-1) groups of Jinhua qingre capsules showed significant inhibitory effect on auricular swelling and significant difference compared with control group (P ＜ 0.01,P ＜ 0.05.);the high dose (10 g· kg-1) and moderate dose (5 g·kg-1) groups of Jinhua Qingre capsules play a marked inhibitory role in the increase in mouse peritoneal capillary permeability (P ＜ 0.01,P ＜ 0.05).In antitussive experiment,high dose (12 g· kg-1) and moderate dose (6 g· kg-1) groups had significant inhibitory effect on cough caused by ammonia water (P ＜ 0.01,P ＜ 0.05) compared with control group.In analgesic experiment,the high dose (12 g·kg-1),moderate dose (6 g·kg-1),and low dose (3 g·kg-1) groups effectively reduced the writhing frequency of mice (P ＜ 0.01,P ＜ 0.05).Conclusion Jinhua qingre capsules have potential effects on anti-inflammatory,antitussive,and analgesic.

OBJECTIVE:To establish the method for simultaneous determination of paeoniflorin,amygdalin,ferulic acid and ligustrazine in Modified buying huanwu decoction.METHODS:RP-HPLC method was adopted.The determination was performed on YMC C18 column with the mobile phase consisted of acetonitrile-0.1％ phosphoric acid (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelengths were set at 320 nm (femlic acid),230 nm (paeoniflorin),207 nm (amygdalin),280 nm (ligustrazine).The column temperature was 30 ℃,and sample size was 10 μL.RESULTS:The linear ranges of paeoniflorin,amygdalin,femlic acid and ligustrazine were 0.191 2-1.912 μg/mL (r=0.999 6),0.117 4-1.174 μg/mL (r=0.999 6),0.011 5-0.115 μg/mL (r=0.999 8) and 0.001 66-0.016 6 μg/mL(r=0.999 7),respectively.The limits of quantitation were 1.912,1.174,0.115,0.016 6 μg/mL,and the limits of detection were 0.25,0.40,0.05,0.008 5 μg/mL,respectively.RSDs of precision,stability and reproducible tests were all lower than 2.0％.The recoveries were 96.9％-100.3％ (RSD=1.3％,n=6),95.1％-100.3％ (RSD=2.2％,n=6),95.3％-100.2％ (RSD=2.0％,n=6)and 97.0％-100.0％ (RSD=1.3％,n=6).CONCLUSIONS:The method is reliable,simple and accurate,and is suitable for simultaneous determination of paeoniflorin,amygdalin,ferulic acid and ligustrazine in Modified buyang huanwu decoction.

Objective To observe the bone marrow mesenchymal stem cells (BMSCs) modified by bone morphogenetic protein-7 (BMP-7) gene on the expression of renal BMP-7, transforming growth factor-β1 (TGF-β1) and vascular endothelial growth factor (VEGF), further to explore its protective mechanism on renal injury in rats with chronic renal failure (CRF). Methods BMSCs with high expression of BMP-7 gene (BMSCs-BMP-7) and empty vector-BMSCs (BMSCs-EV) were obtained by lentiviral-mediated gene transfection. Thirty male Sprague-Dawley (SD) rats were randomly divided into 5 groups, 6 in each group: normal control (CON) group; PBS intervention (CRF with PBS infusion, CRF+PBS) group; BMSCs intervention (CRF with BMSCs infusion, CRF+BMSCs) group;BMSCs-EV intervention (CRF with BMSCs-empty vector infusion, CRF+BMSCs-EV) group and BMSCs-BMP-7 intervention (CRF with BMSCs-BMP-7 infusion, CRF+BMSCs-BMP-7) group. The CRF model was established by 5/6 nephrectomy. The CON group was a sham operation group. The corresponding 12-weeks interventions of each experimental group were performed after 2 weeks of modeling, the rats in the CON group and the CRF+PBS group were injected with 1 ml of PBS through the tail vein, and the other three groups were injected with 1 ml of the corresponding cell suspension once a week. At the time of sacrifice, blood and renal tissue samples were reserved. Serum creatinine (Scr) and blood urea nitrogen (BUN) were measured by routine biochemical methods, and the expression of BMP-7, VEGF, TGF-β1 in kidney was assayed by Western blotting. Results At the time of sacrifice, the levels of Scr and BUN in the CRF+PBS group were significantly higher than those in the CON group (all P<0.01); Compared with the CRF+PBS group, the Scr and BUN of the CRF+BMSCs group, CRF+BMSCs-EV group and CRF+BMSCs-BMP-7 group were decreased to different extents, the differences were statistically significant (all P<0.01);the Scr and BUN of the CRF+BMSCs-BMP-7 group were significantly lower than CRF+BMSCs group and CRF+BMSCs-EV group (all P<0.05). The expression of BMP-7 and VEGF were the lowest in the CRF+PBS group. Compared with the CRF+PBS group, the expression of BMP-7 and VEGF in the CRF+BMSCs group, CRF+BMSCs-EV group and CRF+BMSCs-BMP-7 group were significantly increased respectively (all P<0.05). The expression of the BMP-7 and VEGF in the CRF+BMSCs-BMP-7 group were higher than those in the CRF+BMSCs group and CRF+BMSCs-EV group (P<0.01). Compared with the CON group, the expression of TGF-β1 in the CRF+PBS group was significantly increased (P<0.01);compared with the CRF+PBS group, the expression of TGF-β1 in the CRF+BMSCs group, CRF+BMSCs-EV and CRF+BMSCs-BMP-7 group was significantly decreased (all P<0.01);the expression of TGF-β1 in the CRF+BMSCs-BMP-7 group was lower than the CRF+BMSCs and CRF+BMSCs-EV group (both P<0.01). Conclusions BMSCs modified by BMP - 7 has a protective effect on CRF rats; its protective mechanism may be related to antagonizing TGF-β1 and up-regulation of renal VEGF expression.

OBJECTIVETo investigate the application value of colonic transit test (CTT) combined with anorectal manometry (ARM), barium enema (BE) and defecography (DFG) in accurately evaluating colonic lesions of slow transit constipation complicated with adult megacolon.

METHODSClinical data of 47 above patients admitted between October 2007 and February 2015 in the People's Hospital of Hunan Province were analyzed retrospectively. All the patients were examined with≥2 times of CTT combined with ARM and BE, and 42 cases received additional DFG at the same time, to evaluate colonic lesions before operation. Operative biopsy pathology was used as the standard. The sensitivity, specificity, positive predictive value(PPV) and negative predictive value(NPV) of positioning in the ascending colon and caecum, transverse colon and descending colon were calculated, and the consistency was represented by Kappa test(Kappa value≥0.75 indicates good consistency, meanwhile higher Kappa value indicates better consistency). The Heikkinen score was used to evaluate defecation function at postoperative 6 months.

RESULTSThe age of 47 patients was from 18 to 56 years old. Compared with intraoperative findings and biopsy pathology, the diagnostic coincidence rate was 89.4% by CTT combined with BE and DFG positioning, which suggested pathology-changed colonic segment locating in the ascending colon and cecum (n=12), transverse colon (n=26) and descending colon (n=9), while intraoperative findings and biopsy pathology suggested pathology-changed colonic segment locating in the ascending colon and cecum (n=11), transverse colon (n=23) and descending colon (n=13). The sensitivity was 88.3%, specificity 93.5%, PPV 92.1%, NPV 94.9% and Kappa value was 0.827(P<0.001). Procedures performed included segmental colectomy (n=8), subtotal colectomy (n=29), total colectomy (n=10). There was no serious complication during and after operation. Defecatory function was excellent in 24 cases (60.0%), good in 10 (25.0%), and moderate in 6 (15.0%) evaluated by Heikkinen score at postoperative 6 months. A total of 40 patients were followed up from 1 to 7 years (median 3 years) and there was no long-term diarrhea and recurrence of constipation or giant colon after operation.

CONCLUSIONPreoperative detection of CTT combined with ARM, BE and DFG in patients with slow transit constipation complicated with adult megacolon can make a more precise assessment of the extent of colonic lesions in advance, which has a good clinical predictive value.

Adolescent ; Adult ; Barium Enema ; Cecum ; pathology ; physiopathology ; surgery ; Colectomy ; methods ; Colon ; pathology ; physiopathology ; surgery ; Constipation ; complications ; diagnosis ; pathology ; surgery ; Defecography ; Female ; Gastrointestinal Transit ; physiology ; Humans ; Male ; Manometry ; Megacolon ; complications ; pathology ; surgery ; Middle Aged ; Predictive Value of Tests ; Preoperative Period ; Prognosis ; Recovery of Function ; physiology ; Retrospective Studies ; Sensitivity and Specificity