Objective To analyze and compare the effect,safety and prognosis of early and late digital shaping titanium mesh cranioplasty after skull bone flap defect.Methods One hundred and twelve patients with craniocerebral trauma or cerebral hemorrhage treated by decompressive craniectomy from Jan.2012 to Dec.2013 were included.Seventy-six cases were treated by digital shaping titanium mesh cranioplasty after average 38.9 (32～ 52) d after decompressive craniectomy (early repair group),and thirty-six cases were given operation after 114.2 (90～153) d (advanced repair group).The operation time,skin flap free time,operation blood loss,complications,postoperative patients' activities of daily living (ADL) score,Fugl-Meyery score,neurological deficit score and the recovery of cognitive function of the two groups were compared and analyzed.Results Operation time of early repair group and late repair group were (92.33±13.71) min and (141.67±18.12) min,flap free time were (13.29± 3.43) min and (38.56±7.35) min,and operation bleeding volume were (352.36 ± 21.83) ml and (523.53±32.51) ml (P＜0.01).The subdural effusion rate and the dural rupture rate during skin flap free of early repair group was significantly less than that of late repair group (P＜0.01).ADL and Fugl-Meyery scores in early repair group were increased obviously compared with advanced repair group with significant difference (P＜0.05 or P＜0.01),while the neurological deficit score decreased significantly (P＜0.05).The Montreal cognitive assessment suggested that cognitive function in early repair group was better than that of the late repair group with significant difference (P＜0.05).Conclusions Early digital shaping titanium mesh cranioplasty after skull bone flap defect is better than late treatment with easy soft tissue separation,reduced bleeding and improved prognosis.Digital titanium mesh shaping can ensure precise fit with skull defect,significantly shorten the operation time,and reduce the operation risk.Early digital shaping titanium mesh cranioplasty may have promising clinical application.

Aim To study the transport of geniposide and geniposide in Zhizi Bopi Decoction in MDCK cell membrane model. Methods The safety concentration of geniposide and Zhizi Bopi Decoction in MDCK cells were determined by MTT assay. Then the MDCK cell membrane model was used to investigate the transport of drugs. Firstly, the effects of time, drug concentra-tion, P-gp inhibitor and EDTA on the absorption and transport of geniposide were studied systematically. Secondly, the differences were compared between the transport of the same concentration of geniposide as single compound and that in Zhizi Bopi Decoction in MDCK cell model. The drug concentration was deter-mined by high performance liquid chromatography ( HPLC) to calculate the apparent permeability coeffi-cient (Papp). Results Geniposide transport in MDCK cell monolayer was time and concentration dependent. P-gp inhibitors had no significant effect on its transport and the transport of geniposide was enhanced by ED-TA. The absorption Papp of different concentrations of geniposide in Zhizi Bopi Decoction were ( 8. 96 ± 0. 35 ) × 10 -7 cm · s-1 , ( 8. 95 ± 0. 38 ) × 10 -7 cm · s-1 and (9. 16 ± 0. 30) × 10 -7 cm·s-1, significantly higher than the absorption Papp of geniposide as single compound(5. 85 ± 0. 44) × 10 -7 cm·s-1, (6. 88 ± 0. 38) × 10 -7 cm·s-1 and (6. 31 ± 0. 19) × 10 -7 cm ·s-1 ( P<0. 05 ) . Conclusion The transport of ge-niposide in MDCK cell membrane model is passive transport and is not affected by P-gp. Geniposide may transport via the paracellular route. The Zhizi Bopi De-coction can increase the absorption of geniposide.

Objective To explore the application value of end-to-side closed in situ pancreaticojejunostomy in pancreaticoduodenectomy.Methods The clinical data of 22 patients with obstructive jaundice who were admitted to the Second Affiliated Hospital of Harbin Medical University from January to March 2014 were retrospectively analyzed.All the patients were explored whether tumors invaded inferior vena cava, superior mesenteric vein and portal vein after gallbladder decompression under general anesthesia by tracheal intubation.The standard or extended pancreaticoduodenectomy was applied according to the intraoperative results.The method of pancreaticojejunostomy was end-to-side closed in situ anastomosis of pancreatic duct and jejunal seromuscular layer.The operation time, intraoperative blood loss, postoperative gastrointestinal function recovery time, amylase concentration of drainage at postoperative day 1,3,5, postoperative complication, pathological classification and duration of hospital stay were observed.Patients were followed up by outpatient examination and telephone interview till May 2014.The out-patient follow-up included color Doppler ultrasound examination of effusion near the pancreatic stump, and the telephone interview included whether there were diarrhea of exocrine pancreatic insufficiency.Measurement data with normal distribution were presented as-x ± s (range) , and measurement data with skewed distribution as M(range).Results All the 22 patients underwent successfully the operation, including 17 undergoing standard pancreaticoduodenectomy and 5 undergoing extended pancreaticoduodenectomy, with end-to-side closed in situ anastomosis of pancreatic duct and jejuna seromuscular layer.The operation time of pancreaticoduodenectomy and end-to-side closed in situ pancreaticojejunostomy were (313 ± 37)minutes (range, 228-360 minutes) and(13 ± 4) minutes (7-22 minutes) , respectively.The intraoperative blood loss was (400 ± 207) mL (range, 100-800 mL).The mean tumor size was 3.69 cm2(range, 0.72-1.68 cm2).The recovery time of gastrointestinal function was (5 ±2)days (range, 4-7 days).The serum amylase at postoperative day 1, 3, 5 in the 21 patients was (145±30)U/L (range, 116-180 U/L), (136±40)U/L (range, 105-176 U/L), (147 ±38)U/L(range, 110-175 U/L), and the drainage amylase was (220 ±56)U/L (range, 172-289 U/L), (240 ±54)U/L (range, 192-300 U/L) , (245 ± 52) U/L (range, 190-298 U/L) , respectively.The serum amylase at postoperative day 1, 3, 5 in the patient with pancreatic fistula was 156 U/L, 178 U/L and 177 U/L, and the drainage fluid amylase was 500 U/L, 620 U/L and 605 U/L, respectively.There was 1 patient in the 22 patients with pancreatic duct stent and without death.Among the 4 patients with postoperative complications, 1 patient with grade A postoperative pancreatic fistula recovered after continuous external drainage, the other 3 including 1 case of infection,1 case of pneumonia and 1 of stress ulcer bleeding also recovered after symptomatic and supportive treatment.Postoperative pathological examinations of the 22 patients showed 12 cases of ductal adenoeareinoma, 2 of neuroendocine tumors, 1 of simple cyst, 1 of cystadenocarcinoma, 1 of squamous carcinoma, 1 of adenocarcinoma, 1 of ampullary carcinoma, 1 of tubular adenoma, 1 of leiomyoma and 1 of atypical intraductal hyperplasia.The average length of hospital stay was (11 ±3)days (range, 2-15 days).There were no effusion near the pancreatic stump showed in color Doppler ultrasound examination and diarrhea of exoerine pancreatic insufficiency.Conclusion End-to-side closed in situ pancreaticojejunostomy is safe and feasible, and can be applied to any pancreatic duct size and texture.

Objective:It was a part of an overall study of the clinical research literature about ischemic cerebrovascular disease treated by various acupuncture techniques,namely "restoring consciousness and inducing resuscitation technique","needling Baihui(DU20) through Qubin(GB7)","Jiao’s three-needle technique" and "temporal three-needle technique".The papers collection for analysis were designed on the random,control and blind methods.Methods: A data bank was set up with Access 2003.A statistical analysis was made for the method of employing ratio and constituent ratio about the research literature mamaged with the ramdom,control and blind methods.Results: The ratio of those that were designed randomly is as follows: the "restoring consciousness and inducing resuscitation technique"(50%),"needling Baihui(DU20) through Qubin(GB7)"(100%)(Only two articles meet the request and futher exploration is needed),and the "temporal three-needle technique"(50%).And the "Jiao’s three-needle technique"(40%).The ratio of those that were designed with the control method is as follows: the "restoring consciousness and inducing resuscitation technique"(60%),"needling Baihui(DU20) through Qubin(GB7)"(100%)(Only two articles meet the request and further exploration is needed,the "temporal three-needle technique"(75%),and "Jiao’s three-needle technique"(70%).The blind method was not used.Conclusion: It is suggested that the three methods should be stressed in the study of acupuncture in the treatment of ischemic cerebrocvascular disease.Only in this way,can there be a maximum comparability and the sampling error is decreased.The bias due to subjective factors of the researchers and subjects can be overcome.

Objective To evaluate the safety and efficacy of retroperitoneal laparoscopic resec-tion and reconstructive surgeries in urology. Methods Retroperitoneal laparoseopic resection and re-constructive surgeries were performed on 245 patients including 17 cases of adrenalectomies, 32 cases of radical nephrectomies, 12 cases of partial nephrectomies, 53 cases of nephrectomies, 5 cases of nephroureterectomies, 6 cases of unroofing of peripelvie renal cysts, 46 cases of unroofing of renal cysts, 4 cases of unroofing of polyeystic kidneys, 12 cases of pyeloplasties, 58 cases of ureterolithoto-roles. Results All 245 surgeries were successfully completed. The mean operation time was 59 (20-250) min and the estimated blood loss was 5-300 ml with no transfusion. There was no serious complication during perioperative period. Conclusion Retroperitoneal laparoscopic resection and re-conatruetive surgery in urology is safe and effective with the advantages of minimal invasion, quick re-covery and few complications.

Objective To construct the recombinant plasmid containing human filaggrin gene,purify and identify the immunoreactivity of the recombinant protein,and establish the indirect ELISA to detect AFA for diagnosis of RA.Methods The constructed plasmids were transformed into E. Coli Rosettagami(DE3).This fusion protein was purified by NAT chromatography.ELISA coated with the fusion protein Was established to detect the AFA in serum of patients,which included 114 cases of RA,56 cases of SLE,32 cases of OA and 40 cases of normal controls. The correlation between the results of AFA and anti-CCP in RA group were compared. Results 321 bp fragment of filaggrin gene was amplified and the recombinant expression vector pET-28a( + )-filaggrin was constructed. The sequence of filaggrin gene was the same as the sequence reported in the literatures. The Rosetta-gami (DE3) strains of E. Coli with recombinant vector showed high level of filaggrin protein after induction. The SDS-PAGE showed that the plasmid expressed the filaggrin fusion protein with molecule weight of 14 000 Da. The expression protein could be purified by Ni-NAT with activity. The absorbance value of AFA in RA group was 0.473 ±0. 248 while they were 0. 160 0. 088, 0. 121±0. 070, 0.050 0. ±018 in SLE, OA and normal groups respectively. There were significant differences of absorbance values of AFA between RA and SLE, OA, control group (t = 12.004, 14. 464, 18.078, P<0. 01, respectively). The positivities of anti-filaggrin in RA, SLE and OA were 48.2%, 5.4% and 3. 1% respectively. The positivities of AFA were significantly different between RA, OA and normal control groups (x~2 = 67. 088, P < 0. 01). There was positive correlation of results between AFA and anti-CCP antibody (r = 0.42, P < 0. 05 ) . The consistency rate of results between AFA and anti-CCP was 70. 1%. Anti-CCP was negative in 10 out of 114 patients with AFA positive. AFA can be used to diagnose RA with sensitivity of 48. 2% , specificity of 96.9% , positive predictive value of 93. 2% and negative predictive values of 67. 9% . Conclusions The purified human filaggrin fusion protein is successfully purified. The indirect ELISA method based on the recombinant protein shows good sensitivity and specificity. Joint detection with AFA and anti-CCP can improve the positive rate of detection.

Objective To summarize the clinical characteristics of primary pigmented nodular adrenocortical disease (PPNAD).Methods Eight patients with PPNAD from 2001 to 2009 in PUMC Hospital were reviewed,their clinical data were collected.Results PPNAD often occurred in adolescents.62.5% of patients with PPNAD were complicated with Carney complex(CNC).In addition to general features of Cushing's syndrome,amenorrhea and growth retardation in stature were very frequent in clinical manifestations of PPNAD.Plasma ACTH was undetectable,circadian rhythm of plasma cortisol was disappeared,glucocorticoid excretion was increased paradoxically during the dexamethasone suppression test in 50% patients with PPNAD.Adrenal imaging from 75% patients revealed normal-sized adrenal glands or suspectable micronodular changes.Adrenal pathologic analysis revealed numerous brown cortical nodules containing lipofuscin pigmentation.Unilateral adrenalectomy may relieve symptoms of Cushing's syndrome,but plasma ACTH and circadian rhythm of plasma cortisol were difficult to recovere.Hypercorticoidism might re-occure after unilateral adrenalectomy.Conclusion PPNAD should be bewared in ACTH independent Cushing's syndrome patients without apparent adrenal mass,and CNC should be screened and followed up.

BACKGROUND:Previous studies have shown that monocytes can transdifferentiate into vascular endothelial cells under the induction of various factors including vascular endothelial growth factor(VEGF).It remains poorly understood whether monocytes can be induced to transdifferentiate into lymphatic endothelial cells in vitro.OBJECTIVE:To explore the possibility of the transdifferentiation of monocytes into lymphatic endothelial cells under inflammatory condition.METHODS:Fresh monocytes from peripheral blood were collected by Ficoll density gradient centrifugation and cultured in an endothelial cell medium,followed by incubation in fibronectin-plated well or treated with tumor necrosis factor a for 24 hours,respectively.The expression of specific markers of lymphatic endothelial cells,such as LYVE-1,Podoplanin,Porx-1 and VEGF receptor 3(VEGFR-3),as well as the endothelial cells markers,such as vWF,endothelial nitric oxide synthase(eNOS)and VEGFR-2,were detected by RT-PCR and immunochemical methods.RESULTS AND CONCLUSION:Prior to induction,monocytes were positive to LYVE-1,but negative for Podoplanin,Porx-1,and VEGFR-3,vWF,eNOS,as well as VEGFR-2.Following induction,the cultured mononcytes were positive for Podoplanin,Prox-1 and VEGFR-3,but remained negative for vWF,eNOS and VEGFR-2.It suggested that monocytes can be induced to express the markers of lymphatic endothelial cells stimulated by fibronectin or tumor necrosis factor a.

Aim To study the mechanism of 8-isopro-pylaminomethyl hesperitin ( IPHP ) intestinal absorp-tion using Caco-2 cell lines. Methods Using Caco-2 cell lines as an intestinal epithelial cell model, the effects of drug concentration, temperature, pH, P-gly-coprotein ( P-gp) inhibitor verapamil and multidrug re-sistance protein 2 ( MRP2 ) inhibitors MK-571 or pro-benecid on IPHP transport across Caco-2 cell lines were all investigated. Results The transportation of IPHP was related to drug concentration. The Papp ( AP-BL) ( × 10 -5) was (2. 21 ± 0. 200) cm·s-1,(3. 56 ± 0. 306) cm·s-1,(3. 81 ± 0. 179) cm·s-1,(4. 23 ± 0. 229 ) cm · s-1 , ( 4. 17 ± 0. 262 ) cm · s-1 , re-spectively, and Papp(BL-AP) ( × 10 -5) was (3. 57 ±0. 209) cm·s-1,(4. 51 ± 0. 113) cm·s-1,(4. 97 ± 0. 229) cm·s-1,(5. 24 ± 0. 550) cm·s-1,(5. 07 ± 0. 557) cm·s-1,respectively. Efflux rate was 1. 61, 1. 26,1. 3,1. 23,1. 21,respectively. Temperature and pH both influenced the transport, While the P-gp in-hibitor verapamil had no effect on the transport of IPHP. MRP2 inhibitors MK-571 or probenecid led to an apparent decrease in the efflux of IPHP. Conclu-sion The results suggest that the transport of IPHP is mainly passive diffusion, and MRP2 but not P-gp may be involved in the transport of IPHP.

Aim To observe the effect of melittin on human hepatocelluar carcinoma HepG2 cell prolifera-tion in vitro and its further mechanisms.Methods The capacity of cellular proliferation and apoptosis was measured with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay,Hoechst 33258 assay and Annexin V-FITC /PI assay.The mR-NA expression of Shh, PTCH1, SMO, GLi1 and HDAC2 was performed by qRT-PCR.And the protein expression of Shh,PTCH1,SMO,GLi1 and HDAC2 was assessed by western blotting.Results Our study found that melittin effectively inhibited cell prolifera-tion and promoted cell apoptosis in vitro using MTT method and Flow cytometry.The mRNA and protein expression of Shh,PTCH1,SMO,GLi1 and HDAC2 were obviously decreased after treated with various con-centrations of melittin for 48h in HepG2 cells.Conclu-sions Taken together,our data suggest that melittin could inhibit cell proliferation and promote cell apopto-sis,reduce the level of HDAC2 and down-regulate the Hedgehog signaling pathway in this process simultane-ously.