Objective:To investigate the feasibility of clinical application of HLA-DRB,DPB1 genotyping by PCR-SSP,RFLP for alloge-neic bone marrow transplantation(Allo-BMT) .Methods: Corresponding to each of the DRB alleles, nineteen different pairs of sequence specific primers were used to PCR amplify them from twenty samples in both recipients and donors and their amplified products were directly analyzed on agarose gel. At the same time, a pair of primer was used to PCR amplify the second exon of DPB1 and the restriction fragment length polymorphisms were analyzed on PAGE after their PCR products were separately digested by ten endonucleases. Results: Each DRB was typed in according to its clear and visualized electrophoresis band; the DPB1 genotype was also determined by analysis of all codes representing polymorphism fragment band with a computer. Four couples of recipient and donor were matched. Conclusion:PCR-SSP and RFLP are rapid, accurate and reliable genotyping approaches for Allo-BMT.

Objective: To investigate the dynamic expression of placenta growth factor (PlGF) in the lungs and its role in paraquat-induced pulmonary fibrosis and to evaluate the effect of ACEI captopril and AT (1) -receptor blocker losartan on paraquat-induced pulmonary fibrosis. Methods: 84 adult healthy female Sprague-Dawley (SD) rats were randomly divided into four groups of different treatments designated as: Control, PQ alone (PQ) , captopril treatment, losartan treatment. Each group was divided into three subgroups of seven animals each. The animals were killed at either 7, 14 or 28 days after PQ administration. The rats in PQ group, treatment group were treated intragastrically (ig) with PQ (40 mg/kg) and the rats in control group were treated with the same dose of saline at the beginning of the experiment. The treatment group received Captopril (60 mg/kg; ig) or Losartan (10 mg/kg; ig) once a day respectively after PQ administration and the other two groups received saline. At the given timepoint, animals were sacrificed and lungs were harvested. A semiquantitative assay of histological examination, hydroxyproline in lung tissues were used to determine the severity of alveolitis and fibrosis. RT-PCR and immunohistochemistry were used to detect the mRNA and protein expression of PlGF. Results: Inflammatory cell infiltration and fibrotic scores were more prominent in the model group, hydroxyproline contents in lung tissue were significantly increased after PQ administration compared to the control group. Captopril, losartan apparently attenuated the degree of lung injury and pulmonary fibrosis. On 7th, 14th days, the levels of alveolitis in the intervention groups were significantly alleviated as compared with the model group (P<0.05) . On 28th days, the levels of pulmonary fibrosis in the intervention groups were significantly alleviated as compared with model group (P<0.05) . The hydroxyproline contents in the intervention groups were significantly decreased as compared with model group (P<0.01) . PlGF mRNA on day 7, 14, 28 (1.28±0.29vs0.10±0.01、0.80±0.07vs0.10±0.01、0.65±0.13vs0.10±0.01) in the PQ group were all upregulated as compared with that of the control group. PlGF mRNA on day 7, 14, 28 in the captopril and Losartan intervention groups were significantly decreased (0.94±0.04、0.71±0.09、0.52±0.24 and 0.80±0.12、0.66±0.11、0.51±0.03) . PlGF positive expression index on day 7, 14, 28 (2.27±0.34 vs0.13±0.01、1.78±0.41 vs0.14±0.03、1.25±0.69 vs0.13±0.01) in the PQ group were all upregulated as compared with that of the control group. PlGF positive expression index on day 7, 14, 28 in the captopril and Losartan treatment groups were significantly decreased (1.53±0.78、1.17±0.79、0.97±0.61 and 1.36±0.63、1.24±0.80、0.83±0.47) . PlGF positive expression index on day 7 in the two intervention groups were significantly decreased, as compared with PQ group (P<0.05) . Conclusion PlGF may plays an important role in the development of pulmonary fibrosis following paraquat-induced lung injury in rats. Captopril and losartan had an inhibitory effect on paraquat-induced pulmonary fibrosis, and the effect may be due to inhibition of angiotensin II and, in part, be associated with reduction in PlGF.

Objective To analyze the correlation of hypersensitive C-reactive protein(hs-CRP),homocysteine(Hcy) and D-dimer(D-D) with pathological change degree of coronary heart disease(CHD),and to investigate the diagnostic specificity and sensitivity of joint detection in CHD.Methods A total of 100 cases of CHD patients(experimental group) and 100 healthy subjects(control group) were enrolled,.Patients of the experimental group were divided into four groups on the basis of complications,including simple CHD group(32 cases),amalgamating hypertension group(46 cases),amalgamating diabetes group(9 cases),amalgamating hypertension diabetes group(13cases).The levels of hs-CRP,Hcy and D-D in the five groups were detected and analyzed.Results The levels of hs-CRP,Hcy and D-D in the experimental group were all higher than the control group(P<0.05).Logistic regression confirmed that three indexes were the independent risk factors for CHD.Each indicator has a certain clinical significance to the diagnosis and treatment of CHD but the value of Hcy could be better.Joint detection of hs-CRP and Hcy could be an ideal combination of detection,and the three joint detection might not be suitable for early diagnosis and treatment of CHD.The levels of hs-CRP and D-D in simple CHD group,amalgamating hypertension group,amalgamating hypertension diabetes group were all higher than amalgamating diabetes group(P<0.05).Conclusion hs-CRP,Hcy and D-D could be the independent risk factors of CHD,and joint detection might be with important clinical value for diagnosis of CHD.

Objective To calculate the residoal liver volume using a virtaal liver surgery planning system,and to investigate the value of standardized estimated liver remnant volume ratio (STELR) in prcdicting hepatic dysfunction after hepatectomy.MethodsThe clinical data of 76 patients with primary liver cancer who were admitted to the First Affiliated Hospital of Fujian Medical University from April 2007 to October 2011 were retrospectivcly analyzed.The virtual resection and residual liver volume measurements were carried out using Liv 1.0 software.The value of STELR in predicting hepatic dysfunction was assessed using receiver operator characteristic (ROC) curves.Effects of different risk factors on postoperative hepatic dysfunction were analyzed using univariate analysis of variance and multivariate Logistic regression models. Results The mean residual liver volumes predicted by the software and resected during operation were (489 ± 206)ml and (459 ± 199 )ml,respectively,with a positive correlation between predicted and actual resection volumes (r =0.916,P ＜ 0.05).Of the 76 patients,48 had mild hepatic dysfunction,19 had moderate hepatic dysfunction and 9 had severe hepatic dysfunction.A critical STELR of 53％ was associated with severe hepatic dysfunction.Severe hepatic dysfunction was detected in 2 out of 59 (3％) patients with STELR ≥ 53％ and 7 out of 17 (41％) patients with STELR ＜ 53％,which represented a significant difference ( x2 =5.085,P ＜ 0.05 ).The result of univariate analysis revealed that STEL,R,operating time,intraoperative blood loss were significant prognostic indicators for severe hepatic dysfunction ( F =7.227,8.630,13.809,P ＜0.05).Multivariate Logistic regession revealed that STELR was a significant independent predictor of severe hepatic dysfunction ( Wald =6.675,P ＜ 0.05 ).Conclusion The likelihood of severe hepatic dysfunction after hepatectomy can be predicted by STELR.

OBJECTIVETo establish a real-time quantitative RT-PCR method for the detection of expression of bcl-2 mRNA.

METHODSThe vector containing bcl-2 gene as standard template was constructed with T-A cloning technique. The fluorogenic probe (i.e.,Taq man probe) was used to establish a real-time RT-PCR. bcl-2 mRNA expression level in Burkitt's lymphoma pre-and post-treated with bcl-2 antisense phosphothioate oligodeoxynucleotides (AS-PS-ODN) was determined with real-time quantitative RT-PCR, also with semi-quantitative RT-PCR.

RESULTSThe expression of bcl-2 mRNA in Burkitt's lymphoma treated with bcl-2 AS- PS-ODN decreased significantly and no changes of bcl-2-mRNA expression in group treated with nonsense ODN were noticed. The semi-quantitative RT-PCR results also demonstrated that bcl-2 level varied as detected with real-time fluorogenic quantitative RT-PCR, but less sensitive and accurate.

CONCLUSIONDetection of bcl-2 mRNA expression with the fluorogenic real-time quantitative RT-PCR is more accurate and sensitive than semi-quantitive RT-PCR.

Electrophoresis, Agar Gel ; methods ; Gene Expression ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; RNA, Messenger ; analysis ; Reverse Transcriptase Polymerase Chain Reaction

Objective: To investigate the effect of nucleophosmin 1 (NPM1) mutant A on TGF-β1-induced K562 cell proliferation and AKT phosphorylation. Methods: K562 cells were infected with Ad5-NPM1 to create an NPM1 over-expression cell model. NPM1 levels were determined by ELISA and Western blot analysis. The levels of AKT and P-AKT were determined by Western blot. MTT was used to measure the proliferation of K562 cells. Results: NPM1 protein levels in K562 cells increased in an Ad5-NPM1-MOI-dependent manner. Cell proliferation and NPM1 levels in the supernatant were significantly increased in K562 cells infected with Ad5-NPM1-30 and Ad5-NPM1-100 compared to those infected with Ad5-vector-100 (P<0.01). Treatment with (10 ng/mL) TGF-β1 increased P-AKT levels, but not total AKT levels in K562 cells. TGF-β1-induced phosphorylation of AKT was significantly increased by infection of K562 cells with Ad5-NPM1-100. No significant differences were found in total AKT levels among all groups. TGF-β1 (10 ng/mL) treatment also in-creased the proliferation of K562 cells. TGF-β1-induced K562 cell proliferation was significantly increased by infection with Ad5-NPM1-100 (P<0.01). Conclusions: NPM1 improves TGF-β1-induced cell proliferation by up-regulating AKT phosphorylation levels.

Objective:To explore the structural alterations in functional brain areas of patients with diabetic peripheral neuropathy (DPN) using voxel-based morphometry (VBM), and to investigate the abnormal region of grey matter and its distribution in DPN.Methods:A total of 124 patients with DPN and 88 patients with type 2 diabetes without DPN (NDPN) diagnosed in Haikou Hospital and the First Affiliated Hospital of Guangdong Pharmaceutical University from June 2019 to December 2020 were selected as the study subjects, and 40 healthy volunteers matched with gender and age were included as the control group.All subjects underwent whole-brain MRI examination, and 3D-T1WI data were collected for post-processing and analysis based on voxel morphological analysis.Results:Compared with NDPN patients, decreased gray matter volume in DPN patients was observed in the bilateral anterior central gyrus and thalamus, with statistical signifcant difference ( P<0.05), and there was no significant difference between the two sides ( P>0.05). Compared with healthy control group, decreased gray matter volume in DPN was observed in the the bilateral anterior central gyrus, central posterior gyrus, superior frontal gyrus and thalamus, with statistical signifcant difference ( P<0.05) and there was no significant difference between the two sides ( P>0.05). Conclusions:DPN patients also have decreased volumes of the brain greymatter, suggesting that the occorrence of DPN patients may be caused by the of injury of central structure.

Objective: To explore cyberbullying and risk factors of middle school students, and to provide a reference for cyberbullying prevention in school settings. Methods: A stratified cluster random sampling method was used to select 12 940 students from three junior high schools and four senior high schools in Yixing City of Jiangsu Province, China, to conduct a questionnaire survey from March 1 to May 31, 2019. The Chi -square test was performed to compare differences in the prevalence of cyberbullying among groups with different sociodemographic characteristics, and the multivariate Logistic regression model was employed to analyze the risk factors. A risk predictive nomogram model was constructed and then verified. Results: Middle school students were found to be victims of cyberbullying at a rate of 12.3%. The Logistic regression results showed that alcohol use ( OR =1.93), lack of emotional management ( OR =1.30), feeling unsafe ( OR =1.70), not trusting people ( OR =1.66), increased daily online time ( OR =1.39), higher frequency of using social software or websites ( OR =2.24), poor relationships with family members ( OR =1.46), parental neglect ( OR =1.50), class leadership ( OR =1.30) and poor relationships with classmates ( OR =1.34) were risk factors for middle school students who were victims of cyberbullying ( P <0.05). Based on these 10 independent risk factors, the nomogram prediction model, had good discrimination ( AUC =0.73). Conclusion Cyberbullying is common among middle school students. Internet use, parental neglect and class leadership all have an impact on cyberbullying.

BACKGROUND:Mechanical factors can affect the angiogenic ability of vascular endothelial cells.How the vessel number affects the hydrodynamic properties of microvessels remains to be clarified. OBJECTIVE:To investigate the influence of vessel number on the hydrodynamics of vascular networks based on computational fluid dynamics. METHODS:Three three-dimensional models of vascular network with different vessel numbers were constructed using the Geometry module of ANSYS 19.0 software,and then the vascular network was meshed to tetrahedral elements in Mesh module.The vascular network was assumed to rigid wall without slip,and the blood was assumed to laminar,viscous,and incompressible Newtonian fluid.Blood density,velocity,and a series of blood viscosity coefficients were also established.The Navier-Stokes equation was used for calculation.Hydrodynamic properties of different parts of vascular network with different vessel numbers were analyzed and compared. RESULTS AND CONCLUSION:The streamline,velocity,and mass flow all had the same trend in the vascular network,that is,the outlet and inlet were higher and the middle junction of vascular network was lower.The more the number of vessels,the thinner the blood flow lines in each part of the vascular network.Also,the velocity,mass flow,and wall shear decreased with the increase of the number of blood vessels.Therefore,the changes in vessel number could influence the hydrodynamic environment in the vascular network.Computational fluid dynamics indicates that the changes in vessel numbers can influence the hydrodynamic properties of blood,and provides a new idea for treating bone hypoperfusion-induced diseases(fracture nonunion,bone defect,osteoporosis,etc.)through tonifying kidney and activating blood circulation based on the coupling between angiogenesis and osteogenesis.