Objective To explore the inhibitive effect of BSP on Caspase dependent apoptosis of preosteoclasts RAW264.7 cells by bone sialoprotein (BSP).Methods RAW264.7 cells were treated with the inhibitors of signaling pathway molecules and/or BSP.The activities of caspase3/8/9 in RAW264.7 cells were detected with caspase3/7/8/9 kits and the interaction between Caspase-8 and integrin αVβ3 was detected by Co-IP.Results The activities of Caspase3/8/9 in BSP-treated RAW264.7 cells were decreased and the apoptosis of RAW264.7 cells were inhibited significantly.In addition,there was a positive correlation between caspase-8 activity and caspase-3 activity.There was interaction between caspase-8 and integrin αVβ3.Conclusion The anti-apoptotic effect of BSP on RAW264.7 cells is mainly through the regulation of exogenous apoptosis pathway and the apoptosis induced by caspase-8 is inhibited by the binding of BSP and αvβ3.

Objective To investigate the role of tissue-resident memory T lymphocytes in the pathogenesis of psoriasis.Methods Clinical information was collected from 32 patients with progressive plaque psoriasis.Tissue specimens were obtained from both lesional and nonlesional psoriatic skin of all the patients,as well as from faded lesions in 9 of these patients.Tissue specimens from the normal skin of 10 healthy individuals served as the controls.Immunohistochemical staining was performed to detect the two characteristic surface markers CD69 and CDI03 on tissue-resident memory T lymphocytes and to analyze the status of these T lymphocytes at different stages of psoriasis.The results of immunohistochemical staining were compared by t test.Results The mean number of CD69+CD103+ T lymphocytes per high-power field was significantly higher in lesional skin than in nonlesional skin of the 32 patients (11.34 ± 7.60 vs.2.72 ± 4.20,t =8.46,P ＜ 0.01),but similar between psoriatic lesions in the 9 patients before and after subsidence (14.33 ± 2.21 vs.12.00 ± 4.58,t =1.98,P =0.08).There was no significant difference in the mean number of CD69+CD103+ T lymphocytes between nonlesional psoriatic skin and normal control skin (2.72 ± 4.20 vs.1.70 ± 2.98,t =0.71,P ＞ 0.05).Conclusion Tissue-resident memory T lymphocytes may play a role in the formation and recurrence of psoriatic lesions in patients.

Objective: To analyze the long-term prognosis of re-operation in patients with functional tricuspid regurgitation (FTR) after left sided valve replacement (LSVR) and hence evaluate the optimal timing of mentioned re-operation. Methods: A total of 59 FTR patients who had re-operation after their prior LSVR in our hospital from 1999-01 to 2013-01 were analyzed. The clinical information and post-operative follow-up results were recorded in all patients. Results: There were 5/59 (8.5%) patients died in peri-operative period and the overall post-operative mortality was 11.9% (7/59). The follow-up data of 54 survivors were available for the mean time of 51.1 (21-188) months. There were 19/54 (35.2%) patients suffered from MACE and 30 (55.6%) were beneifted by improved cardiac function. Uni-variable analysis indicated that pre-operative NYHA class IV (P=0.008), pre-operative right ventricular (RV) dysfunction (P=0.037), concomitant left-sided redo-operation (P=0.017) and TVR operation (P=0.002) were associated with all cause mortality of tricuspid re-operation. Multi-variable Cox regression analysis showed that pre-operative RV dysfunction was the only independent risk factor of long term MACE-free accumulating survival rate (HR=3.0, 95% CI 1.11-8.2,P=0.031); while TVR operation (HR=12.8, 95% CI 1.53-107.02,P=0.019) and pre-operative NYHA class IV (HR=5.3, 95% CI 1.20-24.51,P=0.032) were the independent risk factors for long-term mortality in patients after tricuspid re-operation. Conclusion: Patients with compensatory RV function showed better long term prognosis after secondary tricuspid operation. Aggressive re-operation before the occurrence of right ventricular dysfunction could be beneficial for relevant patients.

To simply and rapid authenticate Lonicera japanica. Rapid allele-specific PCR primer was designed base on trnL-trnF 625 G/T Single nucleotide polymorphism and the PCR reaction systems including annealing temperature was optimized; optimized results were performed to authenticate L. japanica and its 9 adulterants. When 100 x SYBR Green I was added in the PCR product of 87 degrees C initial denatured 1 min; 87 degrees C denatured 5 s, 68 degrees C annealing 5 s, 30 cycle; L. japanica visualize strong green fluorescence under 365 nm UV lamp whereas adulterants without. The results indicate rapid allele-specific PCR could authenticate L. japanica and its adulterants rapidly and simply.
Alleles ; DNA Primers ; genetics ; Drug Contamination ; prevention & control ; Lonicera ; classification ; genetics ; Polymerase Chain Reaction ; methods ; Quality Control

ObjectiveTo explore the value of virtual touch tissue quantification (VTQ) in evaluating Lauren classification of advanced gastric carcinoma.MethodsForty-one patients with gastric cancer proved by endoscopic biopsy performed preoperative VTQ examination,and the findings were compared with postoperative pathologic results via hematoxylin -eosin and Alcian Blue-Periodic Acid Schiff (AB-PAS) staining.ResultsIn 41 patients,26 cases were diagnosed as diffuse type and 15 cases as intestinal type by pathology after operation.The shear wave velocity(SWV) of diffuse type was higher than that of intestinal type [(1.72± 0.83)m/s vs (1.05± 0.66)m/s,t =2.819,P=0.008] measured by virtual touch tissue quantification (VTQ).According to the area under the ROC curve,the cut- off value of SWV in gastric cancer tissues for assessing the diffuse type was 1.045 m/s with a sensitivity of 80.8％ and specificity of 73.3 ％ respectively.ConclusionsVTQ could be considered as a promising method to distinguish the Lauren classification in patients with advanced gastric carcinoma.

Objective To investigate the relationship between CYP3A5 gene polymorphism and tacrolimus concentration/dose ratio in children living donor liver transplantation and the correlation with clinical efficacy,for the relatives living donor liver transplantation in children tacrolimus individualized medication providing reference indicators.Methods Peripheral blood samples were collected from children with relatives living donor liver transplantation in the center of liver transplantation,the genotype of CYP3A5 was determined by polymerase chain reaction (PCR)/pyrophosphate sequencing,The dosage of tacrolimus and blood concentration,liver and kidney function and other related indicators were measured within 3 months after operation According to genotypes,the children can be divided into gene expression group (CYP3A5 *1/*1 and CYP3A5 *1/*3) and non-expression group (CYP3A5*3/*3).The drug concentration (C0),tacrolimus dose / body weight (D/W) ratio,drug concentration/dose (C0/D) ratio of each genotype at 1 day,3 d,5 d,7 d,14 d,28 d,2 months and 3 months after administration and the genotype at the time point on liver and kidney function was carried out statistics.Results Among the 80 cases,36 cases (45.0％) were CYP3A5*3/*3,37 cases (46.2％) were CYP3A5*1/*3,7 cases (8.8％) were CYP3A5*1/*1.CYP3A5 gene expression group reached a therapeutic concentration range (C0 ＞ 8 μg/L) than the gene non-expression group takes longer time.There was no significant difference in CYP3A5 gene expression group between the non-expression group on the initial dose (P＞ 0.05);CYP3A5 gene expression group than the non-expression group,tacrolimus C0 within 1 month after operation were statistically significant.CYP3A5 gene expression group than the non-expression group,tacrolimus D/ W in addition to the first day after surgery,other time points were statistically significant (P＜0.05).CYP3A5 gene expression group than the non-expression group,C0/D at the above time points were statistically significant (P＜0.05);There were no significant differences in liver and kidney function between the two genotypes (P ＞ 0.05),but there was significant difference in the alkaline phosphatase.Conclusion CYP3A5 gene expression in children than non-expression group of children need higher doses to reach the therapeutic drug concentration;CYP3A5 gene polymorphism had significant effects on early tacrolimus C0,D/W and C0/D values;CYP3A5 gene polymorphism is instructive for the administration of tacrolimus in children with living donor liver transplantation,CYP3A5 gene type tests should be regular,improve efficacy and reduce the incidence of adverse reactions.

Objective To explore the key signaling pathways and molecules of bone sialoprotein (BSP) to promote proliferation and differentiation of preosteoclasts RAW264.7. Methods RAW264.7 cells were treated with BSP and the inhibitors of signaling pathway molecules. MTS and TRAP staining kits were used to evaluate the effects of proliferation and differentiation. The activity assay kits of Calcineurin, AKT, JNK, ERK and p38 were used to detect their activities. Results Inhibiting ERK and p38 can inhibit cell proliferation induced by BSP significantly, while inhibiting AKT, Calcineurin and PI3K can reduce the role of promoting the differentiation by BSP. With increasing treatment time of BSP, the activity of Calcineurin in RAW264.7 cells increased. Furthermore, the activity of AKT was maximum at 48 h after treated with BSP, while the activity of JNK, ERK and p38 were maximum at 24 h after treated with BSP. Conclusions BSP mainly regulates the ERK and p38 of MAPK pathway to promote RAW264.7 cell proliferation, and regulates AKT, PI3K and Calcineurin pathways to promote RAW264.7 cell differentiation.

The study was purposed to analyze DNA and allele structure of the partial D phenotypes D(Va) and D(VI) of the Rhesus blood group in Chinese. Through polymerase chain reaction (PCR) and direct genomic DNA sequencing, the RHD gene was detected in three weak D individuals identified serologically. The results showed that among the three weak D individuals, one was identified as partial D phenotype D(Va) (Hus) type and genotyped DccEe; another two were testified as D(VI) III type and genotyped DCcee. Moreover, the breakpoints of the replaced region by RHCE in D(Va) (Hus) were 5' end of the exon 5 and 3' end of the intron 5, and there were 7 novel polymorphisms in intron 5: 23-25(GCA)2, 98G>A, 168-169insG, 205-206insT, 494-495insA, 1256-1257insC, 1347G>T. In conclusion the whole exon 5 and intron 5 are replaced by RHCE in D(Va) (Hus) detected in Chinese.
China ; Exons ; genetics ; Genotype ; Humans ; Introns ; genetics ; Phenotype ; Polymerase Chain Reaction ; Rh-Hr Blood-Group System ; genetics ; Sequence Analysis, DNA

Objective To explore a new method for one-stage repair of the intestinal leakage based on the principle of magnetic compression anastomosis. Methods Twenty-four dogs were randomly divided into experimental group (n = 12) and control group (n = 12) according to random number table. The model of upper and multiple intestinal leakages was established by making transverse incisions of 1 cm in length on the jejunum wall about 50 cm and 100 cm away from the Treitz ligament. Forty-eight hours later, two NdFeB magnetic rings with the magnetic flux of 2500 G were put into the intestine from the leak sites. The leak sites were pressed between the two rings. The ventages in the control group were sutured. The condition of the dogs was observed after the repair of the leakage. The excreting time was recorded, and the leakage pressures of the anastomotic stoma were detected.The positions of the magnetic rings in the experimental group were detected by X ray. Tissues of the anastomotic stoma were processed by hematoxylin eosin and Masson staining. All data were analyzed using the two-sample t test. Results Severe abdominal infection occurred 48 hours after the establishment of the model. All the intestinal leakages in the experimental group were successfully repaired and the dogs survived for a long time. The magnetic rings were excreted six or seven days after the repair. Eight dogs of the control group survived. The leakage pressure of the anastomotic stoma seven days after the repair was (134 ±23)mm Hg (1 mm Hg =0. 133 kPa) in the experimental group and (91 ± 18)mm Hg in the control group, respectively, with a significant difference between the two groups (t = 3.225, P ＜ 0.05). The leakage pressure of the anastomotic stoma 14 days after the repair was (281 ±7)mm Hg in the experimental group and (271 ±21) mm Hg in the control group, respectively, with no significant difference between the two groups (t =0. 988, P ＞ 0.05). Histological observation showed that after the magnetic compression anastomosis, the intestinal muscle and mucosa recovered well, inflammatory reaction was slight and less collagen fiber and scar was formed. Conclusions Application of magnetic ring with the magnetic flux of 2500 G in one-stage repair of the intestinal leakage in the state of severe abdominal infection is safe and reliable.

Akt1 is a serine-threonine protein kinase that has been implicated in the control of cellular metabolism,survival and growth.Elevated expression of Akt1 has been noted in a significant percentage of human tumors,promoting cellular metastasis.Conversely,some studies have revealed hyperactivated Akt1 inhibited the invasiveness and metastasis of breast cancer cells.To clarify the definite effect of Akt1 on tumorigenesis and development,Akt1 was silenced by RNAi in the highly metastatic murine breast cancer 4T1 cells.Akt1 silencing didn't affect the proliferation of breast cancer cells in MTT assay,while reduced the migration in Transwell assay.Consistent with the above results,Akt1 silencing didn't change the primary tumor weight,but significantly suppressed lung metastasis of 4T1 cells.These observations indicated Akt1 plays an important role in murine breast cancer metastasis,and suggested that Akt1 might be a therapeutic target for breast cancer metastasis.