Objective To explore the forensic identification points of deaths caused by secondary dam-ages of road traffic accidents. Methods Seventeen deaths caused by secondary dam ages of road traffic accidents were collected. Through scene investigation and necropsy, the basic inform ation of the acci-dents, distribution and property of the injuries, and other inform ation were collected. A ccording to the collected data, the scene was reconstructed in order to confirm the injury process, analyze the way, mechanism and severity of injury, distinguish antem ortem injury from postm ortem injury, and determ ine the cause of deaths. Results C ertain features such as serious injuries, multiple traum as, com bined in-juries, co-existence of antem ortem and postm ortem injuries, multiple causes of wounds, com plex injury mechanism , as well as the mutual dam aging and overlapping injuries were quite characteristically noted in these secondary traffic accident cases. Conclusion Forensic assessment of deaths caused by secondary dam ages of road traffic accidents should be synthetically analyzed and judged through scene investigation and necropsy.

Objective To detect the mouse testicular gene expression pattern differences be-tween spermatogonial stem cell (SSC) proliferative and differential stages and study the molecular reg-ulation mechanism in SSC proliferation and differentiation. Methods With the interval of 24 days, male Kunming mice were injected intraperitoneally with two doses of busulfan (10 mg/kg) to establish spermatogenesis regeneration models. 36 k Mouse Genome Array was used to detect the differential gene expression profiles between the stages of SSC proliferation and differentiation. Bioinforrnsties analysis was conducted in GO (gene ontology) and KEGG (Kyoto Encyclopedia of Genes and Ge-nomes) pathway to describe the potential roles that may play in spermatogonial stern cells behavior regulation. Results Nine hundred and eleven differential expression genes were identified by gene arrays in mice testes, consisting of 608 up-regulated and 303 down-regulated in SSC proliferation stage and SSC differentiation stage. The differential expression genes were classified by their biological process, molecular function and cellular component, respectively. Alterations with statistical signifi-cance (P<0.05)appeared in 84 KEG(;signal pathways, including Notch and Wnt signaling pathways which had been proved to be important for stem cell maintenance. Fifty-six differential expression genes were selected as genes related to stem cells, among which 40 genes were up-regulated, including some stem cell biomarkers(such as Cd9, StraS, hgbl-, Oct4 and Thyl)and some growth factors(such as Fgf2, Pdgfa and Csfl). Conclustion The regulation of SSC proliferation and differentiation involves inmany differentially expressed genes in various signal pathways. This study provides a molecular basis for the elucidation of the molecular mechanism behind self-renewal and differentiation of spermatogonial stem cells.

Objective To explore the underlying clinical factors and precautionary measures of fluid extravasation in patients with calyceal calculi treated by ureteroscopic holmium laser lithotripsy.Methods A retrospective review was made on clinical records of 138 patients with calyceal calculi receiving retrograde ureteroscopic holmium laser lithotripsy from May 2005 to March 2009. The relevance was studied between the occurance of fluid extravasation complications and various clinical factors using x2 test and binary Logistic regression. The clinical factors included patients' sexes, age groups (＜30 years, 30-50 years, ＞50 years), history of treatment (ESWL or open surgery) for upper urinary tract calculi, preoperative upper urinary tract infection, intraoperative placement of ureteral catheter and the length of procedure duration (＜ 50 min, 50-80 min, ＞ 80 min). Results Fluid extravasation complications occurred in 24 patients. The sexes and age groups were irrelevant to the occurance of fluid extravasation complications; while history of ESWL or open surgery and preoperative infection in upper urinary tract, without intraoperative ureteral catheter placement and long duration of procedure were responsible for the higher rates of the fluid extravasation complications.Conclusion Reasonable selection of patients and timing of operation, regular intraoperative ureteral catheter placement and control the length of procedure duration help to reduce fluid extravasation during retrograde ureteroscopic lithotripsy.

BACKGROUND: Vessel extracellular matrix (VECM) is a natural scaffold material obtained from vascular tissues, which can stimulate angiogenesis and accelerate vascularization of tissue-engineered graft, however, the mechanism is poorly understood.OBJECTIVE: To explore the vascularization effects of release of vascular endothelial growth factor (VEGF) from VECM in ureteral reconstitution.DESIGN, TIME AND SETTING: An in vitro cytology observation. The experiment was performed at the Biomedical Engineering Laboratory of First Clinical Medical Science College, Wuhan University, between April and August in 2009.MATERIALS: Abdominal aorta was obtained from 5 rabbits to prepare VECM.METHODS: The VEGF released from VECM in vitro was evaluated by ELISA, the effects of cell proliferation by the released VEGF was detected by MTT colorimetric assay. The defected ureters of rabbits were repaired by homologous VECM in vivo.Then the recovery of the defected ureters and the situation of vasculogenesis were detected at different time point.MAIN OUTCOME MEASURES: The detection of VEGF contents in VECM; and the effects of VECM on vascular endothelial cell proliferation and ureteral reconstitution.RESULTS: In vitro experiment presented that the peak amplitude concentration of VEGF released from VECM in PBS solution was (124.10±1.42) ng/L, which showed proliferative effect on vascular endothelial cells. In vivo, there were some blood vessels on the VECM at 2 weeks after implantation. Epithelial coverage was evident in the lumen of the marginal part of the VECM grafts and the smooth muscle extended from the transition zone. After 8 weeks, the quantity of the blood vessel was increased and the caliber of the blood vessels became wide. There was thickness epithelial lamina in the graft, and the muscle fibers had an organized spatial alignment, forming variably sized bundles. After 16 weeks, there were no significant differences between the regenerative tissue and the normal tissue in morphology.CONCLUSION: The homologous VECM can release VEGF when implanted as tissue engineer biomaterial and might be an ideal replacement biomaterial for ureteral reconstitution.

Objective To establish a long-term culture system for mouse spermatogonial stem cells(SSCs). Methods Testis cells from 4-6 days postpartum male transgenic BALB/C mice were collected by a modified two-step enzymatic digestion method.After three differential adherence selections,the enriched germ cells were finally suspended in StemPro-34 SFM medium supplemented with other nutrients factors and plated on mouse embryonic fibroblast(MEF)feeder layer.20 ng/ml Glial cell line-derived neurotrophic factor,10 ng/ml basic fibroblast growth factor and 200 ng/ml GDNF-family receptor al were added to the serum-free medium to promote SSCs proliferation.Aduh male BALB/C mice,4-5 weeks old,underwent intraperitoneal injection of 40 mg/kg busulfan as recipient mice.Cultured SSCs were also injected into the seminiferous tubules of the left recipient testis through micromanipulator and right testis as self-control.Testes of recipient mice were observed by a fluorescence stereomicroscope and HE stains at 2 months after transplantation. Results By improved digestion method,the vitality of isolated testis cells was more than 98%and the stem cells was enriched about 18.5 fold. 1-2 days after transferred to MEF feeder, the round germ cells started to proliferate and had the shape of paired or aligned undifferentiated spermatogonia connected by cytoplasmic bridges. After 3-4 days, SSCs proliferated continuously and formed typical colonies. SSCs from BALB/c mice could be cultured and passaged in a steady state for 3 months. Cryostat section through the transplanted testis showed that most of seminiferous tubules were filled with germ cells expressing EGFP.HE staining further showed clearly that seminiferous tubules contained complete spermatogenesis.Conclusions SSCs from BALB/c mice could be cultured in an improved culture system for 3 months.The culture system could facilitate understanding the regulatory mechanism that governs SSCs and might provide an opportunity for the cure of infertility.

OBJECTIVE:To determine the oil-water partition coefficients (logP) of rosiglitazone at different pH values by two kinds of methods.METHODS:A shake flask-ultraviolet spectrophotometry method was used to determine plasma concentration of rosiglitazone in water phase and organic phase at variable pH values (0.95,2.0,3.0,4.0,5.07,5.7,6.55,7.06,7.5,8.15,9.21) to calculate logP by using with n-caprylic alcohol-water as simulation system.Above logP values were compared with that calculated by Pallas software.RESULTS:The logP value was lower than 1 when pH value was lower than 4;the logP value was higher than 1 when pH value was ranged from 5 to 9.Measured logP values were in good agreement with that calculated by Pallas software.CONCLUSION:Rosiglitazone exists in the form of hydrochloride salt with good water solubility (pH

Objective To compare the therapeutic effect of retrograde ureteroscopic Ho:YAG laser lithotripsy (UHL) and percutaneous nephrolithotripsy (PCNL) in treating patients with renal calculi of ≤3 cm.Methods From Feb.2008 to Apr.2011,a total of 109 cases (117 renal calculi in total) treated by either PCNL (50) or UHL (67) were retrospectively analyzed.Operative time,stone free rate,postoperative hospital stay,hospitalization expenses and complications were compared in the 2 groups.Results The mean stone burden of the UHL group and PCNL group was 2.5 cm (1.5-3.0) cm and 2.4 cm (1.3-3.0) cm,respectively.There was no significant difference in two groups.In UHL group,operations were performed successfully in all 67 calculi,of whom,43 patients needed combination of flexible ureteroscopy to break the stone fragments falling into the renal calices.No serious complication was recorded except postoperative fever in 3 cases.In PCNL group,all patients had been successful operated.Postoperative fever occurred in 2 cases.Obvious intraoperative and postoperative haemorrhage appeared in 1 case,and cured by selective renal artery embolization.The operation time of the UHL group and PCNL group was (117.0±36.5) min and (90.0±18.3) min respectively,and had no significant difference in two groups (P＞0.05).The stone free rate of the UHL group and PCNL group was 94％ (63/67) and 92％ (46/50) respectively,and had no significant difference in two groups (P＞0.05).Hemoglobin decline of the UHL group and PCNL group was (0.3±0.1) g/L and (20.6±8.1) g/L,and had significant difference in two groups (P＜0.05).Postoperative hospital stay of the UHL group and PCNL group was (5.0±2.4) day and (7.0±3.7) day,and had statistical difference in two groups (P＜ 0.05).Hospitalization expenses of the UHL group and PCNL group was (15 477.0±754.3) RMB and (27 453.0± 1763.5) RMB,and had statistical difference in two groups (P＜0.05).Conclusions Although UHL and PCNL have similar curative effect in treating renal stones of less than 3 cm,UHL have the advantages of lcss trauma and complication,lower expenses and fewer hospital stay.In treating renal stones of smaller than 3 cm,especially in the contraindication of PCNL,UHL may be a better choice.

Objective To compare the dose distribution between three dimensional conformal radiothera -py(3DCRT) and simplified intensity modulated radiotherapy ( sIMRT) for middle-third esopheagal cancer .Meth-ods Twenty middle-third esopheagal cancer patients were enrolled in this study .Dosimetric values for sIMRT and 3DCRT were calculated for total dose of 66 Gy using 2 Gy/day.The parameters included the conformal index (CI),homogeneity index(HI),mean dose and hot spot(V105%)for the planned target volume(PTV)as well as the monitor units(MUs)for plan efficiency,the mean dose(Dmean)for the organs at risk(OAR).The percentage of the normal lung volume receiving≥30,>20 and >5 Gy(V30 Gy,V20 Gy and V5 Gy)were evaluated to deter-mine lung toxity.Results sIMRT achieved significant better Dmin ,Dmean,V105%,HI,CI and spinal cord than 3DCRT(P<0.05).The normal lung V30 Gy and V20 Gy of 3DCRT and sIMRT were 11.61 ±3.68 vs.10.33 ± 3.02(P=0.010)and 23.37 ±8.38 vs.18.89 ±4.45(P=0.001),respectively.Comparable sparing of normal tissue including heart and the normal lung V 5 Gy with both two plans .The number of MU was 542 .24 ±76 .32 for 3DCRT and 530.8 ±97.13 for sIMRT(P=0.677).Conclusion sIMRT provides favorable tumor coverage , homogeneity and conformity and can get better normal lung sparing than 3DCRT.However sIMRT is not superior to 3DCRT in terms of treatment efficiency .

ObjectiveTo investigate the effects of using spiral pedunculated bladder muscle flap ureteroplasty in the treatment of long ureteral segment defects ( ＞ 20 cm).MethodsA retrospective analysis was conducted on the clinical effects of five patients who encountered long ureteral segment defects caused during ureteroscopic lithotripsy.The five patients included three males and two females with an age range from 37 to 59 yrs ( average age 48 ).Four of the cases had defects on the left and one case on the right.Two cases had whole ureteral mucosal avulsion and three cases had whole ureteral ruptur from the pelvis to the bladder junction.Defect lengths measured from 21 to 25 cm( mean length 22.5 cm).All five patients underwent emergency surgery using spiral pedunculated bladder muscle flap ureteroplasty and 7 F double J stent placement in the repaired ureters which was fixed on psoas muscles.The average length of the new ureters using spiral pedunculated bladder muscle flap was 22.5 cm.ResultsAll the operations were successful and the operation time was 1 -2 hrs (average 1.5 hrs).Drainage tubes for four patients were removed three days after operation.IN the remaining case the drainage tube was removed 10 days after surgery due to urine leakage.All wounds healed uneventfully.Serum creatinine and blood urea nitrogen were normal two weeks after surgery.Double-J tubes were removed safely under cystoscope eight weeks after surgery.In following-up,one case was found to have mild hydronephrosis and ipsilateral ureter slight expansion six months after surgery,but renal function was normal.There was no abnomality found in the remaining four patients after 2 -4 years of follow-up.The IVU showed normal morphology and good developments in the ipsilateral ureter.ConclusionsSpiral pedunculated bladder muscle flap ureteroplasty is an ideal treatment method in repairing long ureteral segment defects.

OBJECTIVE To explore the potential targets and mechanisms of the modified Baihe dihuang decoction （MBD/ BDD） applied in post-stroke depression （PSD）. METHODS Network pharmacology was used to mine the potential targets and key pathways of MBD/BDD in the treatment of PSD. PSD model rats were induced by focal cerebral ischemia surgery combined with chronic unforeseen mild stress， and then were randomly divided into PSD model group， MBD/BDD group （12.6 g/kg， by raw drug）， and fluoxetine hydrochloride （FLX） group （positive control， 2.3 mg/kg）； a blank control group was also set up， with 8 rats in each group. Each administration group was given a corresponding medication solution by gavage once a day for 21 consecutive days. The intervention effect of MBD/BDD on depression-like symptoms in model rats was evaluated by open field and forced swimming tests. The brain tissues of rats in each group were dissected and total RNA was extracted for transcriptome sequencing and bioinformatics analysis. The mRNA and protein expressions of genes with significant changes and common neurotrophic factors were verified based on the above results. RESULTS A total of 131 MBD/BDD antidepressant-related target genes were obtained （such as IL1B and AKT1， etc.）， which were closely related to neural active ligand-receptor interactions and cyclic adenosine monophosphate signaling pathway. MBD/BDD could significantly prolong or increase the total time spent and distance traveled in the central grid of qiangzhe@cqtcm.edu.cn PSD model rats， and significantly shorten the cumulative immobility time （P＜0.05）. After treatment with MBD/BDD， the number of genes that changed in rat brain tissue was much higher than that in the FLX group， and there were significant differences in gene profiles among the PSD model group， MBD/BDD group， and FLX group. There were 1 351 differentially expressed genes （DEGs） between the MBD/BDD group and the PSD model group， of which 178 were significantly down-regulated and 1 173 were significantly up-regulated （P＜0.05）. Above 1 351 DEGs were involved in neuronal differentiation， chemical synaptic transmission regulation. They were significantly enriched in axonal guidance， cholinergic synapses and neuroactive ligand-receptor interactions. The top 30 genes in terms of up-regulation in the brain tissue of rats of MBD/BDD group were all associated with neuronal proliferation， development， differentiation， and migration. After MBD/BDD intervention， the expressions of Fezf2， Arx， Ostn， Nrgn genes， brain-derived neurotrophic factor and tyrosine kinase receptor B protein in brain tissue of rats were significantly increased （P＜0.05）. CONCLUSIONS The anti-PSD effect of MBD/BDD may be related to the up-regulation of the expression of genes related to neuronal proliferation， development， differentiation and migration， as well as the promotion of neural structural and functional repair.