Objective To evaluate the effect of the autogenous split-thickness cranial bone grafts on treatment of deformity of oral and maxillofacial regions. Methods The split-thickness cranial bone was harvested through the coronal incisions or the parietotemporal region incisions. The bone graft was then fashioned to the appropriate size and configuration and fixed to the regions of defects and deformities. The cranial bone was used to reconstruct facial bone framework or as sustaining bone graft for facial augmentation. Results 31 patients with the oral and maxillofacial deformities or bone defects were repaired with the cranial bone grafts, including 17 cases of orbital floor defects, 6 cases of malunion of the zygoma fractures, 5 cases of secondary deformity after ankylosis of temporomandibular joint, 2 cases of bone reconstruction after tumor resection and 1 case of cleft palate deformity. The followed-up period ranged from 6 months to 7 years, averaging 11 months. There were no complications of infection and extrusion, no obvious bone resorption was observed, and the facial appearance were greatly improved. Only one patient had a small ectropion which persisted three months. After six-month, the ectropion was not obvious. Conclusion Autogenous bone grafts used to reconstruct the defect and deformity of oral and maxillofacial regions can reduce the risks of infection and extrusion, and there is less visible scar and less bone resorption. Skull bone is an ideal source of bone graft material in the oral and maxillofacial deformity.

Objective To investigate the effects of rhubarb on the expression of glucocorticoids receptor (GR)and peripheral blood lymphocytes in burning-induced septic rats. Methods Sixty-six male healthy Sprague-Dawley(SD)rats were randomly divided into sham operated control group(n=18),sepsis model group(n=24) and rhubarb treatment group(n=24),each group was further randomly divided into 12,24 and 72 hours subgroups according to different time points. The model of scald sepsis was replicated by scald injury induced by boiling water at the rat back accounting for 30% total body surface area(Ⅲ grade of scald),and administration of endotoxin (5 mg/kg)into the peritoneal cavity 12 hours after scald injury. After the successful establishment of septic models, the rats in the rhubarb treatment group were immediately infused with 50 mg/kg rhubarb powder dissolved in 1 mL saline through a gastric tube,while the rats in sham operated control group and sepsis model group received saline by the same way as a substitute for rhubarb. The the binding capacity of GR of peripheral blood leucocyte and binding activity of GR of hepatocyte were analyzed by radiation ligands binding assay. The CD4+,CD8+as well as CD4+/CD8+ ratio in peripheral blood lymphocytes were detected by flow cytometer. Results The binding capacity of GR of peripheral blood leucocyte and binding activity of GR of hepatocyte were significantly decreased in a time-dependent manner in sepsis model group compared to those of the sham operated control group,while in the rhubarb treatment group they were increased in a time-dependent manner after interference of rhubarb, and they were higher than those in the model group at the same time points〔leukocyte GR binding capacity (locus/cell)at 12,24,72 hours :1 515.38±300.44,1 859.63±258.26,1 890.50±307.88 vs. 1 122.63±225.39, 1 008.88±150.41,724.38±91.19;hepatocyte GR binding capacity(fmol/mg):210.19±26.26,258.01±20.98, 283.38±38.21 vs. 153.11±30.07, 129.83±26.89, 94.08±14.30, all P<0.01〕. Compared with the sham operated control group,the CD4+ and CD8+ were decreased in various degrees at 12 hours and 24 hours in the septic group, at 24 hours the differences being statistically significant (P<0.01 and P<0.05). CD4+/CD8+ratios were decreased significantly at all time points,the differences were statistically significant at 24 hours and 72 hours(both P<0.01). The CD4+ T cell and CD4+/CD8+ ratio at all the time points were increased at various degrees in the rhubarb treatment group,and the differences from those in the sepsis model group at 24 hours and 72 hours were statistically significant (1.58±0.69, 1.56±0.49 vs. 1.02±0.41, 1.01±1.68, both P<0.01). Conclusion Rhubarb can modulate the binding capacity of GR of peripheral blood leucocyte and the binding activity of GR of hepatocyte,and via its influence on the number of peripheral leucocytes,the immune dysfunction in the sepsis processes is improved.

[Objective] To screen and identify the new Y-STR loci from the Y chromosome and examine the polymorphism of these Y-STR loci. [Method] To seek and locate the position of 5 Y-STR loci, including DYS709, DYS720, DYS721, DYS722, and DYS723, and perform sequencing of these 5 Y-STR loci. Then to investigate the polymorphism in unrelated Chinese Han males. [Results] Five Y-STR loci were identified from Y chromosome sequence. By scrutinizing the physical position on Y chromosome of previously reported Y-STRs, we found that three loci were novel and two loci overlapped with two loci published only online. All loci could be male-specifically amplified with a product size ranging from 185 bp to 278 bp. After 108 males of the Chinese Han Population (Guangzhou) were examined, we found 5 DYS709, 11 DYS720 alleles, 4 DYS721 alleles, 6 DYS722 alleles, and 6 DYS723 alleles. A total of 95 haplotypes were identified, 84 of which were unique, and with a haplotype diversity of 0.997 2±0.001 2(HD±SE). [Conclusion] This set of Y-STRs can be used as Y chromosome genetic makers in related fields.

Objective To investigate Toll-like receptor-4 (TLR4) protein expression in human pancreatic adenocarcinoma,and to evaluate the relationship between TLR4 protein expression and angiogenesis.Methods Sixty-two surgically resected human pancreatic adenocarcinoma specimens and 35 normal para-cancerous tissues were investigated for TLR4 protein expression by immunohistochemical SP methods,and CD31 antibody was used to mark microvascular endothelial cells and determine the microvessel density (MVD).The correlation among TLR4 protein expression and MVD and clinicopathologic features of pancreatic adenocarcinoma were analyzed.Results TLR4 protein positive expression rate and MVD in human pancreatic adenocarcinoma was 74.2％ (46/62) and 47.3 ± 13.5,respectively,which were significantly higher than those in the normal pancreatic tissue [17.1％ (6/35),12.6 ±4.8; P ＜0.01].TLR4 protein positive expression rate in the cases with lymph node metastasis was 83.8％,which was significantly higher than that in the cases without lymph node metastasis (60.0％,P =0.036).TLR4 protein positive expression rate in the patients with stage Ⅲ and Ⅳ of TNM classification was 85.3％,which was significantly higher than that in the patients with stage Ⅰ and Ⅱ (60.7％,P=0.028).MVD was closely related to tumor size,lymph node metastasis and TNM stage of pancreatic adenocarcinoma (P =0.008,0.036,0.010).There was a strong positive correlation between TLR4 protein expression and MVD (r =0.534,P ＜0.01 ).Conclusions TLR4 protein expression is closely related to the development and progression of human pancreatic adenocarcinoma and its potential mechanism is related to the promotion of tumor angiogenesis.

Objective To observe the effect of triptolide combined with gemcitabine on proliferation and apoptosis of pancreatic cancer cells ,and to analyze the relevant mechanisms .Methods After treated with TPL ,GEM or TPL combined with GEM in vitro , PANC-1 cells proliferation was accessed by MTT assay and the interaction between the two drugs was calculated .Apoptotic mor-phological changes and apoptosis rate of the cells were investigated by Hoechst 33258 staining and flow cytometry ,respectively .The expression of signal transduction and transcription factor 3(STAT3) ,cysteine aspartate specific proteases-3(caspase-3) protein were detected by Western blot analysis .Results TPL ,GEM or TPL combined with GEM could significantly inhibit the prolifera-tion of PANC-1 cells ,and the combination of the two drugs had a synergistic effect .The cells of the TPL group ,GEM group ,as well as the combined group showed typical apoptotic morphological changes .Compared with the TPL group and GEM group ,the number of apoptotic cells of the combined group increased significantly .Compared with the control group ,the cells apoptosis rate of the TPL group ,GEM group and combined group was significantly increased (P< 0 .05) ,and the apoptosis rate of the the combined group was significantly higher than that of the monotherapy group(P<0 .05) .TPL combined with GEM synergistically inhibited p-STAT3 protein expression and activated caspase-3 protein expression .Conclusion TPL combined with GEM can synergistically in-hibit proliferation and induce apoptosis of pancreatic cancer PANC-1 cells ,its mechanism is related to the inhibition of STAT 3 sig-naling pathway ,promotion the expression of caspase-3 protein .

Objective To observe the relationship between Toll-like receptor 4 (TLR4) and the sensitivity of PANC1 cells to gemcitabine (GEM),and to analyze the potential mechanism.Methods PANC1 cells were divided into GEM group,lipopolysaccharide (LPS) + GEM group and TLR4-siRNA + GEM group.GEM group was treated by GEM alone.LPS + GEM group was pretreated with 1 mg/L LPS for 4 h and then treated by GEM.TLR4-siRNA + GEM group was transfected with 100 pmol/mL TLR4-siRNA for 4 h and then treated by GEM.The untreated cells were used as the control group.MTT method was used to detect the cell proliferation.Morphological changes and apoptosis rate of the cells were examined by Hoechst33258 staining and flow cytometry,respectively.The protein expression of TLR4,phosphorylated AKT (p-AKT) and activated Caspase-3 were detected by Western blot.Results The median inhibition concentration (ICs0) of GEM in the GEM group,LPS + GEM group and TLR4-siRNA + GEM group was (8.9 ± 0.32),(14.21 ±0.95),(3.96 ± 0.27) mg/L,respectively.The IC50 in LPS + GEM group was significantly higher than that in GEM group (P ＜ 0.01),and the IC50 of GEM in TLR4-siRNA + GEM group was significantly lower than that in GEM group (P ＜0.01).Compared with that in GEM group,the cells with typical apoptotic morphological changes were decreased in LPS + GEM group,which was increased in TLR4-siRNA + GEM group.The apoptotic rate in control group,GEM group,LPS + GEM group,TLR4-siRNA + GEM group was (2.1 ± 0.3) ％,(15.1 ± 2.3) ％,(9.8 ± 1.5) ％,(22.9 ± 3.1) ％,respectively.Compared with that in GEM group,the cells apoptotic rate was significantly reduced in LPS + GEM group (P ＜0.01),which was significantly increased in TLR4-siRNA + GEM group (P ＜0.01).TLR4 protein level in the 4 groups was 0.83 ±0.08,0.81 ±0.07,0.85 ±0.07 and 0.16 ±0.03;p-AKT protein level 0.61 ±0.05,0.36 ±0.03,0.73 ± 0.07 and 0.21 ± 0.02;activated Caspase-3 protein level was 0.66 ± 0.05,0.73 ± 0.07,0.45 ± 0.04 and 0.91 ± 0.07,respectively.The expression of TLR4 and p-AKT in TLR4-siRNA + GEM group was significantly lower than that in GEM group (P ＜0.01),while the expression of activated Caspase-3 protein was increased significantly (P ＜ 0.05).Compared with the GEM group,the expression of p-AKT protein in LPS + GEM group was significantly increased (P＜0.01),and the expression of activated Caspase-3 protein was significantly decreased (P＜0.01).Conclusions TLR4 can inhibit the sensitivity of pancreatic cancer PNAC1 cells to GEM,and the mechanism is related to the activation of PI3K/AKT pathway and downregulation of activated Caspase-3.

Objective To evaluate the effect of chronic low potassium on K+uptake rate in the my?ocardium and skeletal muscle of rabbits. Methods Thirty?two adult male rabbits, aged 12-14 weeks, weighing 2?0-2?7 kg, were randomly divided into 4 groups ( n=8 each) using a random number table:normal feeding group ( group N) , low potassium feeding group ( group L) , potassium supplementation con?trol group ( group SC ) and potassium supplementation experimental group ( group SE ) . N and SC groups were given a normal diet only, and L and SE groups were fed with a low potassium diet for 15 days. Potassi?um chloride ( KCl) 0?5 mol∕L was then infused intravenously at the initial rate of 60 μmol·kg-1 ·min-1 in SE and SC groups. Blood samples were obtained from the central artery of the left ear every 5 min for meas?urement of plasma K+ concentrations. The infusion rat of KCl was then adjusted until the plasma K+concen?tration reached 5?5 mmol∕L and maintained at this level for 1 h, and then infusion was stopped. The total volume of KCl infused was recorded. The hearts and soleus muscle of animals were excised for determination of K+content. K+uptake and uptake rate were calculated. Results Compared with N group, the plasma K+concentration, and K+content in the myocardium and soleus muscle were significantly decreased in group L ( P<0?05) . Compared with SC group, the total volume of KCl infused, and K+uptake and uptake rate in the myocardium and soleus muscle were significantly increased in group SE ( P<0?05) . Conclusion Chro?nic hypokalaemia can increase K+ uptake rate in the myocardium and skeletal muscle of rabbits.

[Objective] To propose a criterion for making conclusions on paternity tests based on STR genotyping. [Method] To use binomial distribution formula to calculate minimal numbers of STR loci that must be tested for different scenarios in paternity testing. [ Results ] We proposed a set of criteria for making STR paternity testing conclusions. For triplet tests, concluded "paternity positive" for the following four cases when the cumulative paternity index (PI) was greater than 10 000: 1) no inconsistent STR locus was detected in 15 loci (PE > 0.571 4/locus) or 2) only one inconsistent STR locus was detected in 19 loci or 3) only two inconsistent STR loci were detected in 28 loci or 4) only three inconsistent STR loci were detected in 35 loci; otherwise, concluded "paternity negative" when at least four inconsistent STR loci had been detected. For single parent tests, concluded "paternity non-exclusive" for the following cases when the cumulative PI was greater than 10 000: 1) no inconsistent STR locus was detected in 18 loci (PE>0.411/locus) or 2) only one inconsistent STR locus was detected in 29 loci or 3) two inconsistent STR loci were detected in 41 loci; concluded "paternity negative" when three or more inconsistent loci were detected. [Conclusion] Our experience has proven that these criteria are robust in STR paternity testing.

Objective To estimate the validity of Charlson' s weighted index of comorbidities (WIC) used to predicting 28-day mortality among ICU pneumonia patients with underlying diseases.Methods Aretrospective analysis of 160 adult patients with pneumonia admitted to a multi-discipline ICU of Shanghai Changzheng hospital between October 2010 and February 2012 was carried out.Clinical data were collected including age,gender,community acquired pneumonia (CAP) or hospital acquired pneumonia (HAP),underlying diseases,severity-of-sepsis,and 28-day mortality.WIC scores,acute physiology and chronic health evaluation (APACHE) Ⅱ,and sepsis related organ failure assessment (SOFA) were assessed within the first 24 hours after admission.Logistic regression analyses were used to evaluate the predictors for outcome.The receiver operating characteristic curve (ROC) was used to compare the performance of these scores between different methods.Results Of 160 enrolled patients,76 (48.8％) were CAP,82 (51.2％) HAP,and 106 (66.3％) male,54 (33.7％) female,and 99 (61.9％) patients survived and 61 (38.1％) died.The average age was (62.4 ± 17.3) years old.Compared with survivors,WIC,APACHE Ⅱ and SOFA scores were significantly higher in death group (P ＜ 0.05).The multivariate logistic regression revealed that risk of death depends predominantly on age (OR =1.049,95％ CI:1.011-1.088,P =0.011),WIC (OR =1.725,95％ CI:1.194-2.492,P =0.004),APACHE Ⅱ score (OR =1.175,95％CI:1.058-1.305,P =0.003),SOFA score (OR =1.277,95％ CI:1.048-1.556,P =0.015),presence of ARDS (OR =0.081,95％ CI:0.008-0.829,P =0.034),and complicated with severe sepsis (OR =0.149,95％ CI:0.232-0.622,P =0.004).The area under the receiver operating characteristics curve in predicting mortality was 0.639 (0.547-0.730) for the WIC,0.782 (0.709-0.856) for APACHE Ⅱ score,0.79 (0.714-0.866) for SOFA score and 0.842 (0.777-0.907) for the merger of three.Conclusions In pneumonia patients of ICU,WIC is a useful approach to predicting 28-day mortality,and the risk of death significantly depends on co-morbidities.