BACKGROUND: Above curative dose, there is a dose-dependent correlation of the survival rate of fat transplantation with vascular endothelial growth factor (VEGF), the curative dose of VEGF determines the tissue volume of transplanted fat. OBJECTIVE: To investigate the changes of the blood supply rebuilding and the survival rate of the fat transplant in the microenvironment of continuous infusion of VEGF using osmotic pump. DESIGN, TIME AND SETTING: A cellular morphology observational experiment was performed from September 2007 to October 2008 in Department of Plastic Surgery at the Affiliated Hospital of Guangdong Medical College. MATERIALS: Ninety healthy SD rats were used to establish autologous fat transplantation models. Human VEGF was produced by American ONCOGEN Corporation. Type Alzet2002 mini-osmotic pump was produced by American Alzet Corporation. METHODS: Ninety rats were equally divided into 3 groups randomly. The operation control group was only processed into autologous fat transplantation; Experimental control group received autologous fat transplantation and continuous infusion of 2 000 ?L physiological saline using osmotic pump at a speed of 0.5 ?L/h for 14 days; Experimental group received autologous fat transplantation and continuous infusion of 2 000 ?L physiological saline containing 1 ?g VEGF using osmotic pump at a speed of 0.5 ?L/h for 14 days. MAIN OUTCOME MEASURES: At the 7th, 14th, 28th, 42nd, 64th days postoperation, the graft’s survival volumes were measured using mini-scale test tube; the cell morphology changes were observed with hematoxylin-eosin stain; The newly born microvessels on experimental group and control groups were researched with immunohistochemistry staining. RESULTS: At 14 days postoperation, the graft’s survival volumes of experimental group was higher than operation control group and experimental control group (P 0.05). At each time point, the vascular density of the fat transplant in the experimental group was higher than that in other groups (P 0.05). CONCLUSION: VEGF can speed up vasifaction and raise fat graft survival significantly.

Objective To compare the curative effect,patient's satisfaction degree,and adverse reaction on seborrheic keratosis by level-cutting and frozening with liquid nitrogen.Methods 68 cases were randomly divided into liquid nitrogen frozening group(32 cases,group A)and flat-cutting group (36 cases,group B).Liquid nitrogen was spurted until freeze-thaw status in group A,and the ramnant skin lesion needed for other treatments,but no more than 3 times.The skin lesion in group A was completely resected with scalpel following its basilar part after local anesthesia,and elaeoyarn was pressurized to stop bleeding on wound,which was removed afer 5 days.The effects,satisfactory degree and adverse reaction were analyzed after 3 mouths of treatment.Results The effect and satisfactory degree in group B was significantly better than that in group A(P＜0.01),but the adverse reaction in group B was significantly lower than that in group A(P＜0.05).Conclusion Compared with liquid nitrogen frozening,the therapy of flat-cutting has advantages to seborrheic keratosis,with good efficacy,high patient's satisfaction and low adverse reaction.

Objective To investigate the effects of Pollen Typhae total flavone (PTF) on INS-1 pancreaticβcell damage induced by palmitic acid ( PA) . Methods INS-1 pancreatic β cells were given long-term induction with PA to establish the impaired cell model, and then were intervened with PTF. Cell viability was determined by tetrazolium salt ( XTT) colorimetry. Results PA impaired the viabilities of INS-1 pancreatic β cells in concentration- and time-dependent manner, and PTF improved the impairment of INS-1 pancreatic β cells induced by PA in concentration -dependent manner. Moreover, PTF showed better improvement on the impairment when the INS-1 pancreatic β cells were impaired more seriously by PA. Conclusion PTF has effects on ameliorating the impairment of INS-1 pancreaticβcells induced by PA for long time.

Objective To investigate the bone marrow supernatant expression of sclerostin in the patients with multiple myeloma (MM) and to clarify its clinical signification .Methods The sclerostin level was quantified by using ELISA ,and the gene expression of sclerostin was determined by RT-PCR .Results The sclerostin level was (0 .54 ± 0 .21)pg/mL in the MM group ,which was sig-nificantly higher than (0 .31 ± 0 .06)pg/mL in the control group (t=5 .67 ,P<0 .01) .The sclerostin level was (0 .65 ± 0 .17) pg/mL in the recurrent and refractory MM group ,which was significantly higher than (0 .47 ± 0 .21) pg/mL in the control group and the newly diagnosed group (t=8 .44 ,3 .27 ,P<0 .01) ,RT-PCR verified that the BMMNC of most patients expressed sclerostin gene .The expression of sclerostin in the MM group was negatively correlated with alkaline phosphatase (ALP)(r= -0 .379 ,P=0 .005) ,and positively related with the correction blood calcium ,bone loss points ,serum β2-micro globulin(β2-GM ) ,proportion of serum M protein and clinical International Staging System (ISS) stages .The median follow-up periods were 29(6-65) months ,the low sclerostin group had the median survival period of 48(6-65) months and the high sclerostin group had the median survival pe-riod of 24(6-52) months ,the difference between them had statistical significance (χ2 = 12 .74 ,P< 0 .01) .Conclusion Its level may reflect the bone destruction ,osteogenesis inhibition degree and myeloma burden ,and reflect the median survival period of MM patients to some extent .

Objective To explore the relationship between HLA?B allele polymorphisms and nasopharyngeal carcinoma ( NPC) in Xinjiang, China and its clinical significance. Methods A total of 226 patients were assigned to NPC group, while 207 healthy volunteers were assigned to control group. PCR amplification with sequence?specific primers was used to determine HLA?B alleles. Comparison of HLA?B allele frequency between the above two groups, between Han and Uygur populations, and between patients with various clinical characteristics of NPC was made by chi?square test. The Kaplan?Meier method was used to calculate the survival rates and the log?rank univariate analysis was used to explore the relationship between survival rates and HLA?B allele frequency. Results In all the subjects or Han population alone, the allele frequency of HLA?B?46 in the NPC group was significantly higher than that in the control group ( P=0. 000;P=0. 000 ) . In Uygur population, however, there was no significant difference in the allele frequency of HLA?B?46 between the NPC group and the control group (P>0. 05). In the patients with NPC, those less than 30 years old had a significantly higher allele frequency of HLA?B?44 than those no less than 30 years old (P=0. 029);those with differentiated non?keratinizing carcinoma had a significantly higher allele frequency of HLA?B?48 than those with undifferentiated non?keratinizing carcinoma ( P=0. 029);those with stage T1+T2 disease had a significantly higher allele frequency of HLA?B?48 than those with stage T3+T4 disease ( P=0. 029) . The 5?year overall survival, disease?free survival, distant metastasis?free survival, and local relapse?free survival rates had no relationship with the expression of HLA?B?46, HLA?B?44, or HLA?B?48 in NPC patients ( all P>0. 05) . Conclusions HLA?B?46 allele is probably a NPC susceptibility gene in Han population in Xinjiang. HLA?B?44 is probably associated with early age of onset, while HLA?B?48 is probably associated with the pathological type and T stage of NPC. Therefore, HLA?B alleles are probably associated with the development and progression of NPC.

Anti-angiogenic drugs such as bevacizumab can effectively alleviate the patients′brain edema and clinical symptoms and improve the patients′life quality by reducing vascular permeability and the blood-brain barrier damage.Bevacizumab has got positive efficacy in the clinical,so that it is considered as an effective and safe treatment for malignant brain edema.

Objective:To screen and characterize the common epitopes recognized by monoclonal antibody 3A8,which binds several kinds of Gram negative bacteria,which were recognized by monoclonol antibody 3A8 agaisnt different strains of LPS.Methods:Phage clones binding to 3A8 were screened from C7C phage displayed peptide library,the attached phage clones were eluted by LPS2630(O111:B4).After 3 rounds of panning,the positive phage clones were identified by ELISA,and the amino acid sequences of these positive clones were deduced from DNA sequences.In order to predict how these peptide mimics interaction with 3A8,peptides(A2 and A5) based on conserved sequences of positive clones were synthesized and conjugated with KLH for further study.The binding of A2-KLH and A5-KLH to 3A8 were identified by ELISA.Results:15 of 33 phage clones were identified as positive clones and shown specific binding with 3A8.LPS2630 potently inhibited the binding of phage clone to 3A8.In analysis of the amino acid sequence,there were seven kinds of sequences containing highly hydrophilic residues,and Ser Pro Pro/Pro X Pro was the conserved sequence.The peptides A2 and A5 could bind to 3A8 specially.Conclusion:The conserved sequences containing Ser Pro Pro/Pro X Pro are obtained,which are recognized by mAb 3A8 against broad spectrum Gram negative bacteria and several strain LPS.The synthetic peptides based on the conserved sequence can bind to 3A8,indicating that the peptides can mimic a common epitope of Gram negative bacteria,which be expected as candidate epitope for vaccinization.

ObjectiveTo investigate the expression of CCL2 and its effects on the proliferation and adhesiveness on leukemia cells in patients with acute lymphoblastic leukemia(ALL). MethodsThe bone marrow mesenchymal cells (BMMSC) and bone marrow mononuclear cells (BMMNC) of 30 ALL patients and 30 healthy controls were studied, and CCL2 level was evaluated by ELISA. CCL2 gene mRNA level in ALL was determined by RT-PCR. The cell proliferation and adhesiveness were detected by MTT assays. The cell counts were measured by flow cytometry. ResultsThe BM plasma levels of CCL2 in patients at diagnosis were significantly higher than that in healthy controls [(780.12±129.61) pg/ml vs (120.49±25.21) pg/ml,t =4.96, P =0.00]. Supernatant levels of CCL2 in BMMSC were significantly higher than that of BMMNC [(572.38±35.39) pg/ml vs (193.85±15.45) pg/ml,t =5.37,P =0.00]in vitro.CCL2 cannot induce leukemia proliferation alone,but could induce leukemia proliferation in BMMNC and BMMSC co-culture in a dose- and time-dependent manner.CCL2 could increase the leukemia adhesive to the BMMSC compared with control (r =0.824,P =0.02).ConclusionPatients with B type ALL had higher levels of CCL2 which was secreted by BMMSC. The leukemia could induce the BMMSC to secrete CCL2. CCL2 could promote the survival and proliferation of leukemia in the presence of BMMSC and BMMNC, and enhance ALL cells adhesion toBMMSC in a dose-dependent manner.

Objective To evaluate the advantages of cytomegalovirus (CMV) real-time PCR, to monitor CMV infection in a population of transplantation recipients in comparison with the qualitative PCR method.Methods 150 plasma samples and leukocytes samples collected from 59 bone marrow transplant recipients and 9 liver transplant recipients were tested by qualitative CMV PCR assay in parallel; Real-time CMV PCR using Roche Light Cycler was performed with 150 plasma samples and 54 control plasma samples.Results Using real-time PCR as the reference standard, the sensitivity and specificity of the qualitative CMV PCR assay with leukocytes samples were 62.5% and 95.5%, with plasma samples were 57.5% and 99.1% respectively. The recipients with the level of CMV DNA over 5?10~3 copies/ml by real-time PCR had higher percentage of developing CMV disease than those below 10~3 copies/ml.Conclusion The quantitative detection of CMV DNA by real-time PCR with plasma is a rapid, specific and sensitive method to monitor CMV infection in patients after transplantation and to guide antivirus therapy.

Objective:To investigate the germline mutation status in multi-pathway in Chinese female breast cancer patients and explore their correlation with clinicopathological characteristics. Aim to enrich the database of breast cancer germline gene mutations in Chinese population and provide laboratory evidence for the application of breast cancer targeted drugs.Methods:From January 2017 to July 2019, whole blood samples were collected from 148 women (age of onset concentrated in the 24~80 years old) diagnosed pathologically with breast cancer in the Department of breast surgery, Peking University People′s Hospital. Germline mutations in HR, MMR, BER, and KDR pathway related genes were detected by next-generation sequencing. The pathogenicity interpretation was performed, and pathogenic, likely pathogenic, and mutations of uncertain significance were screened. The clinicopathological characteristics including age at the onset, luminal typing, tumor size, metastasis, and family history were analyzed, and the correlation between mutations in different pathway genes and clinicopathological characteristics was analyzed by the Chi-squared test and Fisher′s exact probability test.Results:Among the 148 patients, there were 69 cases of HR mutations (including three types of mutations, including pathogenic, likely pathogenic and uncertain significance), 16 cases of MMR mutations, 6 cases of BER mutations and 8 cases of KDR mutation. ATM mutations in the HR pathway were associated with luminal typing ( P=0.054), and patients with HER2+breast cancer were more likely to carry ATM mutations. PMS2 mutations in the MMR pathway were correlated with tumor size ( P=0.060), and patients with tumor size>50 mm were more likely to carry PMS2 mutations. KDR mutations was significantly correlated with luminal typing and family history. ( P=0.021, P=0.024). Conclusion:The mutation frequency in BER, KDR, MMR and HR pathways in Chinese breast cancer patients increased successively. Germline mutations in ATM, PMS2 and KDR genes may be involved in the development of breast cancer in the Chinese population. Multi-pathway gene detection of breast cancer can provide laboratory evidence for the use of PARP inhibitors, trastuzumab and other targeted drugs.