Objective To explore the distribution of parvovirus B19 (HPVB19) infection in patients with leukopenia.Methods Patients who visited the Affiliated Hospital of Hangzhou Normal University from January 2015 to June 2016 were analyzed.Patients with peripheral leucocytes count less than 3.5×109/L were included in experiment group and healthy people were included in control group.HPVB19 IgG and IgM were detected by enzyme-linked immunosorbent assay, and HPVB19 DNA was detected by quantitative polymerase chain reaction.Differences in continuous data between two groups were compared with two-sample t test and those in categorical data were compared with Chi-square test.Results A total of 79 patients were included in experiment group, including 32 males and 47 females.Ages ranged from 24 to 62 years old.And 126 healthy individuals were included in control group, including 55 males and 71 females.Ages ranged from 28 to 67 years old.The positive rates of HPVB19 IgG, IgM and DNA in experiment group were 34.2%, 5.1% and 3.8%, respectively, while those in control group were 36.5%, 0 and 0, respectively.The detection rates of HPVB19 IgM and DNA between two groups were significantly different (χ2=6.507, P=0.011 and χ2=4.856, P=0.028, respectively).Sequence analysis for 3 of the HPVB19 DNA positive samples showed that there were two single nucleotide polymorphisms in VP1/VP2 sequence from one patient, which contributed to the 153rd (L/H) and 219th (N/Y) amino acids mutations, respectively.Phylogenetic analysis found that two strains belong to genotype 1a and one strain belongs to genotype 1b.Conclusions Detection rate of parvovirus HPVB19 infection (positive rates of HPVB19 IgM and DNA) in leukopenic patients is significantly higher than healthy controls.HPVB19 should be detected before considering transfusion in leukopenic patients in clinical practice.

Objective To investigate renal expression of ApoE in human lupus nephritis (LN) and its relationship with renal damages.Methods Renal expression of ApoE was examined in 40 LN patient biopsy samples by immunohistochemieal staining.The relationship between ApoE levels" and renal pathologic changes,renal injuries was analyzed.Results Only weak ApoE expression was found in normal renal tissue.The expression of ApoE was significantly higher in LN patients [ glomerular:(31.2±11.0) vs (3.0±1.6);tub-ular:(71.64±8.90) vs (0.60±0.06)].The expression of ApoE was higher in class IV than non-class IV LN [glomerular: (38.4±4.5) vs (18.0±4.6);tubular: (77.6±2.5) vs (60.0±3.2)].Spearman's relation-ship analysis showed that the expression of ApoE in glomerular and tubular was closely related to the renal histological score activity index and functional changes in LN (P＜0.01).Conclusion The expression of ApoE is high in renal tissues of LN,especially in class IV,and the high expression may be associated with LN activity and progressive renal dysfunction.

Objective To explore the invasion effect of CXCR3 overexpression on T lymphoblastic leukemia (Jurkat cells) with chemokine receptors. Methods Mouse CXCR3 was amplified by RT-PCR and overexpressing CXCR3 lentivirus carrying GFP&Puromycin (puro) was constructed. CXCR3 expression on infected Jurkat cells surface was detected by FCM. Constructed cells were seeded in Transwell invasion model to study whether CXCR3 overexpression would increase the invasion or not. Results GFP expression on Jurkat cells was less than 10% after 96 h lentivirus infection. CXCR3 expression was 90% higher than vector group , and GFP expression reached 90% after screening. Therefore, Jurkat cells with stable overexpression of CXCR3 were successfully constructed. Invasion rate of Jurkat CXCR3 cells was [(12.71 ± 1.03)%], which was significant higher than that of vector control group [(6.82 ± 0.49)%], (P < 0.0001). Conclusions CXCR3 expression on leukemia cells is closely associated with leukemia invasion. The increase of CXCR3 expression can enhance the invasion of leukemia cells, and may be one of the mechanisms of T lymphoblastic leukemia invasion.

Objective To analyze the advantage of altered fractionation radiotherapy by calculating the accumulative effects of daily biologically effective dose (BED) to find out the difference between conventional fractionated radiotherapy and altered fractionation radiotherapy.Methods The data in the report of hyperfractionated or accelerated radiotherapy for head and neck cancer published by Cochrane Collaboration in 2010 was analyzed.Based on the radiotherapy processes mentioned in this report,the accumulative effects of daily BED were calculated and compared in different radiotherapy processes by using linear-quadratic mode.The variation of BED in different radiotherapy processes was find out.Results In total dose of unity as the premise of 70 Gy,altered fractionation especially the hyperfractionated accelerated radiotherapy could give a higher BED to the tumor during a shorter period,hyperfractionated radiotherapy could give a lower BED to normal tissues,and hyperfractionated radiotherapy with split course could give higher BED to the tumor while lower BED to normal tissues.Conclusions The variation of BED in different radiotherapy processes can be shown clearly by linear-quadratic mode.It can be simple and shortcut through mathematical models for the evaluation of different radiotherapy plan,on clinical symptomatic selection play a guiding role in tumor therapy.

It is known that ischemic cerebrovascular disease is causing enormous harm to human health on account of the resultant high morbidity and disability rate. In this connexion, the anticipated target is to control the size of focal ischemic cerebral infarction, which is also an important method for judgment of therapeutic efficacy. The key question is to survey the size accurately and objectively; at the same time, the quantitative analysis of focal ischemic cerebral infarction is the pivotal question affecting the experiment conclusion and the reliability level. In this paper are introduced and summarized the methods being recently and commonly used in survey and computation, and the studies made on quantitative analysis of focal ischemic cerebral infarction by 2, 3, 5-triphenyltetrazolium chloride (TTC) staining method. Also are summarized the principles of dyeing in TTC method, the preparatory work, and the commonly used method of surveying and computation. It is the intent of this review to provide relevant data and suggestion for research workers.
Animals ; Brain Ischemia ; pathology ; Cerebral Infarction ; pathology ; Coloring Agents ; Humans ; Reperfusion Injury ; pathology ; Tetrazolium Salts

BACKGROUND: The understanding about characteristics and variation trends of cerebral vascular hemodynamic indexes in high-risk stroke population,in prior and post-stroke period patients as well as in normal people,particularly the cerebral vascular hemodynamic index(CVHD) prior to stroke can improve diagnosis efficiency of stroke and increase sensitivity and specificity of stroke diagnosis.OBJECTIVE: To study the features of CVHD in acute stoke,in order to provide evidences for early evaluation and rehabilitation intervention.DESIGN: Case analysis and observational study based on patients.SETTING: A neurological department in a general hospital of military area command of Chinese PLA and a prevention and cure institute of cerebrovascular diseases.PARTICIPANTS: Totally 55 (male 38,female 17) inpatients in the Neurological Department of Fuzhou General Hospital of Chinese PLA from June to December 2001 were enrolled in the study.METHODS: CVHD was measured during the first week after onset and the data were analyzed.MAIN OUTCOME MEASURES: The CVHD,blood pressure and cerebral vascular hemodynamic parameters.RESULTS: The CVHD of each patient is below 75,median 32.5. The mean velocity,blood flow,maximum velocity,minimum velocity and difference between diastolic and critical pressures were lower than corresponding ones of healthy men in seventies( P ＜ 0.01 ). And the pulse wave velocity,peripheral resistance,specific impedance and critical pressure increased( P ＜ 0. 01).CONCLUSION: During the first week after stroke,the CVHD decreases and many hemodynamic parameters alters significantly. Examination of CVHD can reveal cerebral circulation insufficiency or predict an attack of cerebral stroke.

Objective To determine the expression of CD4+ CD25+ FOXP3+ regulatory T cells(Treg cells) in peripheral blood of patients with hepatocellular carcinoma (HCC) and investigate the clinical significance of Treg cells determination in clinical practice. Methods Flow cytometry was employed to measure the levels of CD4+ CD25+ T cells and CD4+ CD25+ FOXP3+ T cells in peripheral blood of 18 HCC patients, 26 hospitalized patients without HCC (clinical control) as well as 24 healthy persons (healthy control). Results The percentage of CD4+ CD25+ T cells in total CD4+T cells isolated from the HCC patients(4.25% ± 3.98 % ) was elevated significantly compared to that in the clinical control group (1.34% ± 1.14%) or healthy control group (1.29% ±0.95%) (both P＜0.01). There was no difference in the percentage between the clinical control group and the healthy control group (P＞0.05). Meanwhile, the ratio between CD4+ CD25+ FOXP3+ T cells and CD4+ T cells in HCC patients (2.94%0.91%) also increased significantly compared to that in the clinical control group (0.76% ± 0.34%) or healthy control group (0.81% ± 0.29%) ( both P＜0. 001), which showed a more obvious increasing tendency than the ratio of CD4+ CD25+ T cells and CD4+ T cells. No difference in percentage of CD4+ CD25+ FOXP3+ T cells in CD4+ T cells was found between the clinical control group and the healthy control group (P＞0.05). Conclusion As the more accurate regulatory T cells, CD4+ CD25+ FOXP3+ T cells are able to detect the increase of population in HCC patients. Therefore, it is important to determine the levels of CD4+ CD25+FOXP3+ T cells in HCC patients for prevention and treatment of malignancy.

Objective To explore the differential liver plasma membrane (PM) proteins that may be related to the occurrence,development and reversal process of hepatitis and to understand the pathogenesis of hepatitis and the new drug targets by performing a comparative proteomics research of liver PM between hepatitis B surface antigen (HBsAg) transgenic mice and wild-type C57 mice.Methods A 6-month-old HBsAg transgenic mouse model was established.The pathological examination was performed to observe the pathological changes of transgenic mice and wild-type C57 mice.The PM from liver tissue of 6-month-old transgenic mouse and the control mouse were purified through twice sucrose density grade centrifugation combined with second antibody magnetic bead enrichment.The purity of extracted PM was verified by Western blot.Differential proteome expression analysis was performed by using two-dimensional electrophoresis (2-DE) and ImageMaster software analysis.The differentially expressed proteins were lysed by trypsin and identified by liquid chromatography combined with tandem mass spectrometry (LC-MS/MS) analysis.Results The pathological examination results showed that hepatitis was observed in the transgenic mouse group,while no abnormity was found in the controls.The PM was successfully enriched and the mitochondria contamination was reduced by sucrose density grade centrifugation combined with second antibody magnetic bead purification treatment.Thirty differential mice liver PM protein spots were visualized,in which 11 non-redundant proteins were successfully identified by LC-MS/MS in transgenic mouse group,including 9 up regulated protein spots and 2 down-regulated protein spots.These differentially expressed proteins included keratin,cardiac Ca2+ release channel,cytochrome B5,ATP synthase subunit alpha,etc.Conclusions A batch of HBsAg gene expression related differential proteins are identified in mouse liver plasma.These proteins might be new drug targets for anti-HBV treatment.This study will guide further investigation on the mechanism of HBV infection induced hepatitis.

Objective To explore the value of the laparoscopic ultrasonography (LUS) in laparoscopic common bile duct explora-tion(LCBDE) .Methods 47 cases with cholecystolithiasis combined with bile duct dilatation or abnormal liver function were tested by conventional ultrasound ,magnetic resonance cholangiopancreatgraphy (MRCP) CT and LUS after LCBDE .All results were com-pared with the surgical findings and the intraoperative biliary endoscopy results .Results The diagnosis coincidence rate of conven-tional ultrasonography ,CT ,MRCP and LUS in 47 cases were 85 .1% ,72 .3% ,95 .7% ,100 .0% respectively .6 cases completed the surgery under the guidance of LUS ,1 case was converted to open surgery .Conclusion LUS is an examination method to avoid bile duct injury effectively and reduce postoperative residual stone in LCBDE .

Objective To investigate the role of the p38 MAPK pathway in the formation of cytoplasmic vacuoles .Methods Af-ter treated with Anisomycin ,SB203580 or SP600125 ,images of HepG2 ,LM3 ,QBC939 ,Hela and A549 cells were recorded by light microscopy and taken at a magnification of 400 × .The effects of anisomycin ,SB203580 and SP600125 on the activity of p38 and JNK were measured by Western blot .LM3 and A549 cells were stained with the ER-tracker red and the lyso-tracker red and subjec-ted to confocal microscopy analysis .Results (1)Anisomycin could abolish cytoplasmic vacuolization of HepG2 cells .(2)p38 MAPK activation was responsible for anisomycin-induced cytoplasmic vacuolization abolishment .(3)p38 MAPK blocking initiated cytoplas-mic vacuoles formation in various cancer cell lines .(4)p38 MAPK blocking-induced cytoplasmic vacuoles disrupted the integrity of endoplasmic reticulum .(5)p38 MAPK blocking reversibly induced cytoplasmic vacuoles formation .Conclusion These observations provide direct evidence for a role of p38 MAPK signaling in regulating the formation of cytoplasmic vacuoles .