Effects of Chronic Salt Loading on Plasma Inflammatory Factors in Normotensive Adults / 中国循环杂志

Objectives: To observe the effects of dietary sodium intake on plasma inflammatory factors including tumor necrosis factor alpha (TNF-α), high-sensitivity C-reactive protein (CRP) and monocyte chemoattractant protein -1 (MCP-1) in normotensive adults. Methods: Thirty normotensive volunteers, aged 18 to 60 years old, were selected to undergo baseline survey, low-sodium diet (51.3 mmol per day) for 7 days, followed by high-sodium diet (307.8 mmol per day) for 7days. Subjects were classified as salt sensitive (SS, 10 subjects) or non-salt sensitive (NSS, 20 subjects) based on their mean arterial blood pressure (MAP) increase (SS: more than 10 percent increase at the end of the high-sodium phase compared with the end of low-sodium phase). Fasting blood samples were taken on the first day of baseline and on the sixth day of the two intervention phases. Plasma TNF-α and MCP-1 concentration was measured using an enzyme-linked immunosorbent assay system, plasma hs-CRP concentration was measured by immune nephelometry. Results: The prevalence of SS is 33%. After salt loading, no significant change was found in the plasma hs-CRP concentrations; Whereas plasma TNF-α level increased significantly in both of the SS and NSS groups(pg/ml, [168.4±67.8 vs 42.1±26.7], P<0.01 and [129.8±24.1 vs 37.7±15.8], P<0.01, respectively) ; Plasma MCP-1 was also significantly higher during the high-sodium than the low-sodium phase in both SS and NSS groups(pg/ml, [205.2±64.2 vs 166.3±48.5], P<0.01and [212.3±52.2 vs 143.6±55.9], P<0.01). Conclusions: High-sodium diet can induce an inflammatory state independent of the salt sensitivity in normotensive subjects.

The influence of familial factors on post rabies exposure vaccination in rural habitants / 中华流行病学杂志

Objective To understand the current situation of potential exposure to rabies among the rural habitants in Hunan province,and to study the impact related to familial factors on post rabies exposure vaccination.Methods In total,40 villages from 20 townships of 4 counties were selected by multistage sampling method.Study samples were selected from these villagers and familial basic information and vaccination post rabies exposures were recorded through questionnaires.Data were statistically analyzed by SPSS 17.0.Results Among 3042 villagers from 864 households being surveyed,124 person-time exposures were found from January,2009 to October,2010,with a total exposure rate as 4.08％,and the annual average exposure rate as 2.33％.Data from univariante analysis showed that the rates on post rabies exposure vaccination were statistically correlated with the following four factors:knowledge on the score of rabies prevention (x2 =8.260,P =0.042),whether being involved in the new type of rural cooperative medical care (P =0.035),family disposable cash income in the year of 2009 (x2 =10.831,P =0.031),distance between the households and the health facilities in towns and townships (x2 =9.071,P=0.033).Results from logistic regression analysis indicated that the score of knowledge on rabies prevention (OR=1.420,95％CI:1.055-1.905) and the annual disposable cash income of the family in 2009 (OR=1.480,95％ CI:1.044-2.098) were independent factors that influencing the rabies vaccination.Conclusion Strengthening the education programs on rabies prevention in rural habitants and increasing family income were feasible way to increase the rate of rabies vaccination in rural areas of Hunan province.

Serological survey of pandemic influenza A (H1N1) virus infection among population in Hunan Province / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To understand the infection condition and analytical methods of Influenza A (H1N1) virus in the population of Hunan Province during different periods.</p><p><b>METHODS</b>Quick surveys on the positive rate of Influenza A (H1N1) virus hemagglutination inhibition (HI) test have been conducted for 5 times successively from November 2009 to March 2010 in 14 medical and health institutions of Changsha city, whose results were then compared with those from the sampling surveys of whole Hunan province.</p><p><b>RESULTS</b>2131 subjects were involved in this study; the total population standardized rates of antibody positive investigated for 5 times were 9.32% , 14.62%, 31.08%, 28.43% and 22.80% respectively; the population of 6-17-years-old has the highest rate of antibody positive; only 9.84% of the antibody positive subjects attributed to vaccine inoculation; there was no significant difference in the standardized positive rates between the quick serological surveys and the corresponding sampling survey of Hunan province (P > 0.05).</p><p><b>CONCLUSION</b>The positive rate of A (H1N1) virus antibody reached the peak in late January 2010; quick investigations in small region could be used to evaluate the infection prevalence during pandemic of infectious diseases.</p>

Histology and proliferative capability of thoracic vertebral body growth plates of rats at different ages / 南方医科大学学报

<p><b>OBJECTIVE</b>To compare the histological features of the thoracic vertebral body growth plates (VBGPs) of rats at different ages and assess their proliferative capability.</p><p><b>METHODS</b>The thoracic VBGPs obtained from rats aged 1 day and 1, 4, 8, 16 and 28 weeks were identified using safranin O-fast green staining, and the height of the hypertrophic zone, proliferative zone, and resting zone were measured. The chondrocytes were isolated from these VBGPs with a modified trypsin-collagenase type II digestion method for primary culture in vitro. The expressions of proliferating cell nuclear antigen (PCNA) mRNA and protein was detected by real time-PCR and Western blotting, respectively.</p><p><b>RESULTS</b>The 1-day- and 1-week-old rats showed significantly greater hypertrophic zone and proliferative zone in the VBGPs than older rats (P<0.01); the proliferative zone was significantly greater in rats aged 4 weeks than in those aged 28 weeks (P<0.05). The resting zone was obviously greater in rats aged 1 day and 1 week than in older rats (P<0.05), and also greater in rats aged 4 weeks than in those aged 16 and 28 weeks (P<0.05). Obvious ossification in the resting zone occurred at 16 weeks, and most of the resting zone became ossified at 28 weeks. The expression of PCNA decreased at both the mRNA and protein levels as the rats grew.</p><p><b>CONCLUSION</b>The 3 zones of VBGPs are greater in rats aged 1 day and 1 week than in older ones. Ossification in the resting zone begins at 16 weeks, and till 28 weeks, most of the resting zone is ossified. The proliferation ability of VBGP chondrocytes decreases with the increase of age of the rats.</p>

Risk factors of death cases of hand-foot-and-mouth disease in Hunan province / 中华预防医学杂志

<p><b>OBJECTIVE</b>To study risk factors of death cases of hand foot and mouth diseases (HFMD) in Hunan province, so as to provide scientific evidence for further prevention and control.</p><p><b>METHODS</b>The 105 death cases of HFMD between January and October, 2010 in Hunan Province were selected as case group; and the 210 survival cases of serious HFMD, which were matched by gender and resident places with a ratio at 2:1 in the same period in Hunan were selected as control group. The basic information, hospitalized experience and previous medical history had been surveyed and the relevant risk factors were analyzed by single factor and multi-factor logistic regression.</p><p><b>RESULTS</b>In case group, 79.05% (83/105) of the cases lived in rural area and 9.52% (10/105) of the cases lived in urban-rural midst area. In control group, 87.62% (184/210) of the cases lived in rural area and 11.43% (24/210) of the cases lived in urban-rural midst area. In case group, 59.05% (62/105) of the patients first visited rural (private) clinics and 20.00% (21/105) first visited community hospitals in villages and towns; while in control group, 43.81% (92/210) and 13.33% (28/210) chose rural (private) clinics and community hospitals in villages and towns as the first choice respectively.22.86% (24/105) of the case group and 39.05% (82/210) of the control group were diagnosed as HFMD in their first visit to hospital.27.62% (29/105) of the case group and 7.14% (15/210) in control group were provided pyrazolone in the treatment. For glucocorticoid, 80.95% (85/105) and 5.71% (6/105) of the case group were given as treatment by rural (private) clinics and community hospitals in villages and towns separately; while the proportions in the control group were 41.43% (87/210) and 0.48% (1/210) respectively. For antibiotics, 35.24% (37/105) and 23.81% (25/105) of the case group were prescribed by rural (private) clinics and community hospitals in villages and towns separately; while the percentages in the control group were 15.71% (33/210) and 7.14% (15/210). 3.81% (4/105) of the case group and 11.90% (25/210) of the control group were vaccinated in one month before the onset. The results of single-factor logistic regression indicated that living in rural areas (OR = 0.075, 95%CI: 0.016 - 0.343) and in rural-urban midst areas (OR = 0.069, 95%CI: 0.013 - 0.368), diagnosis of HFMD in the first visit to hospital (OR = 0.463, 95%CI: 0.271 - 0.788) and vaccination one month before the onset (OR = 0.293, 95%CI: 0.099 - 0.866) were four protective factors; while rural (private) clinics as the first choice (OR = 4.717, 95%CI: 1.891 - 11.767), community hospital in villages and towns as the first choice (OR = 5.250, 95%CI: 1.883 - 14.641), medication of pyrazolone (OR = 4.961, 95%CI: 2.520 - 9.766), medication of glucocorticoid in rural (private) clinics (OR = 6.009, 95%CI: 3.435 - 10.510) and in community hospital in villages and towns (OR = 12.667, 95%CI: 1.505 - 106.638), medication of antibiotics in rural (private) clinics (OR = 2.918, 95%CI: 1.690 - 5.040) and in community hospital in villages and towns (OR = 4.062, 95%CI: 2.036 - 8.108) were seven risk factors. The results of multi-factors logistic regression showed that medication of pyrazolone (OR = 2.311, 95%CI: 1.062 - 5.030), medication of glucocorticoid in rural (private) clinics (OR = 5.480, 95%CI: 3.039 - 9.880), medication of antibiotics in rural (private) clinics (OR = 2.430, 95%CI: 1.301 - 4.538) and medication of antibiotics in community hospitals in villages and towns (OR = 3.344, 95%CI: 1.477 - 7.569) were the risk factors of death of HFMD.</p><p><b>CONCLUSION</b>The risk factors of HFMD deaths include the medication of pyrazolone, glucocorticoid and antibiotics by rural (private) clinics and medical institutions in villages and towns. The department concerned should revise the technical manual to standardize the medication of the above drugs.</p>

Surveillance on the etiology and genetic characteristics of rabies in Hunan province, from 2008 to 2009 / 中华流行病学杂志

Objective To analyze the etiology of rabies in Hunan province and the genetic characteristics of rabies N gene isolated from 2008 to 2009.Methods Direct immunofluorescence assay (DFA) and nested PCR were employed to detect the monitoring samples including brain tissues of dogs and saliva,serum or urine which were collected in 2008 to 2009,from the rabies patients.Positive samples were sequenced by ABI3730 gene analyzer for the full length of the N gene target.The homology and hpylogeography of the rabies virus were analyzed after the phylogenetic tree was constructed by Blast,Clustal W and Mega 4.0 software.Results Of the 1451 tissue samples from the dogs' brain,31 were positive under DFA and the positive rate was 2.14％.The DFA positive samples were redeteeted by RT-PCR and the positive rate was 1.17％.56 samples of saliva,serum and urine samples were detected by RT-PCR from the rabies patients,with 3 positives and the positive rate was 5.36％.The length of nest PCR products were 255 bp.The rates of homology to the nucleotide and the amino acid of rabies N gene were 87.2％-87.9％ after compared to the pasture strain.The phylogenetic tree was successfully built and 20 strains isolated lately belonged to the rabies gene type Ⅰ.Conclusion The epidemic situation of human and dogs rabies in Human were relatively stable,with all the isolated rabies virus belonging to genotype Ⅰ,without any variation.

Effects of anti-TGF-beta1 monoclonal antibody on in vitro expansion of cord blood CD34(+) cells / 中国实验血液学杂志

In order to investigate the effect of anti-TGF-beta1 monoclonal antibody on the expansion of cord blood CD34(+) cells, the purified cord blood CD34(+) cells were divided into three groups: blank control group: purified cord blood CD34(+) cells cultured on day 0; control group: cells cultured for 3 days in the culture system, containing SCF, IL-3, IL-6 and FLT3-L; test group: cells cultured for 3 days in the same culture system as control group, but with anti-TGF-beta1 monoclonal antibody. The mononuclear cell counting (MNC), the expression of CD34 and c-kit, and CFU-GEMM, BFU-E and CFU-GM counting were detected in all three groups. The result showed that the expansion of MNCs, CD34(+) cells and CD34(+)c-kit(+) cells in test group [(2.35 +/- 0.25) x 10(5), (1.16 +/- 0.29) x 10(5), (1.09 +/- 0.26) x 10(5)] was significantly higher than that in control group [(1.25 +/- 0.13) x 10(5), (0.55 +/- 0.19) x 10(5), (0.51 +/- 0.2) x 10(5)](p < 0.01). The expansion of more primitive CD34(+)c-kit(-) subpopulation in test group [(12.95 +/- 3.17) x 10(3)] was even significantly higher than in control group (1.71 +/- 0.83) x 10(3) (p < 0.01). Colony forming assay showed that the number of earlier progenitor colony CFU-GEMM, BFU-E in test group [(16.3 +/- 4.72) x 10(3), (60.5 +/- 20.96) x 10(3)] was higher than that in control group [(5.0 +/- 2.58) x 10(3), (16.25 +/- 7.93) x 10(3)] (p < 0.01). The number of relatively mature CFU-GM between test group and control group was not statistical significance [(6.33 +/- 2.85) x 10(3) vs (4.0 +/- 2.28) x 10(3)](p > 0.05), but both were higher than that in blank group (0.75 +/- 0.29) x 10(3). These results demonstrated that anti-TGF-beta1 monoclonal antibody promoted the expansion of MNC and CD34(+) cells, and even more marked expansion of the more primitive progenitor cells-CD34(+)c-kit(-) cells. Meanwhile, it enhanced the output of more immature colony CFU-GEMM and BFU-E, but had no evident influence on the mature myeloid colony CFU-GM. It is concluded that the anti-TGF-beta1 monoclonal antibody can synergize other cytokines to enhance the proliferation of cord blood CD34(+) progenitor cells effectively, and it is more important that can reserve more primitive progenitor cells.

Pathogenic diagnosis of an unexplained epidemic event of infectious diarrhea / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To determine the pathogen of an unexplained epidemic event of infectious diarrhea by laboratory diagnosis of suspected cases samples.</p><p><b>METHODS</b>28 samples from 28 suspected cases (22 fecal samples, 3 vomitus samples, 3 anus swab samples) were tested for Norovirus by RT-PCR. Sequencing and phylogenetic analysis were acomplished of 5 positive samples.</p><p><b>RESULTS</b>160 of 5694 population were ill with an attack rate of 2.81%. The peak period was 7-9, March. 14 of 28 samples were tested Norovirus positive.Sequencing and phylogenetic analysis showed Norovirus type GII/4 was the causative agent and it had highest identity (97. 9%) with epidemic strain 2006b.</p><p><b>CONCLUSION</b>The epidemic event ofinfectious diarrhea were caused by GII/4 Norovirus strains.</p>

Viral surveillance on rabies in Hunan province,in 2006 / 中华流行病学杂志

Objective To understand the source and distribution of rabies virus(RV)in Hunan province with viral surveillance in order to provide scientific measures for prevention and control on rabies.Methods Brain samples from healthy-looking domestic dogs were collected in the agricultural markets at the dist6cLs of high.middle,and lOW incidence rates and detected by direct Immunofluorescence assay (DFA).Positive samples would be further detected by RT-PCR and the surveillance samples were detected bv RT.PCR.The positive samples detected bv RT-PCR were sequenced with N gene.Results The infection rate of thosc healthy-looking domestic dogs with rabies virus was 2.78％in Hunan province in 2005.23 positive samples’N gene were sequenced and their similarities were 88.8％-100.0％.The results indicated that Hunan rabies virus N gene aberrance was mainly synonymous aberrance and did not CKITy obvious regional characteristics.The rabies virus were circulating among different districts in Hunan province,and the neighboring provinces such as Guizhou,Hubei，GuangxiComparison of immunity to measles between floating and local population,Jiangsu and Henan.There were no positive samples detected in salivary，blood and urine samples.There was one positive sample detected in two skin samples.Conclusion There are dogs infected with rabies virus found in Hunan province and this study showed that rabies virus detected in Hunan had a close genetic relationship with those rabies idcntified in other provinces,suggesting that study on the immunity and management of dog related rabies should be strengthened.

Relationships of serum selenium, VEGF and sFas levels in leukemia patients / 中国实验血液学杂志

In order to investigate the relationships of cancer chemopreventive trace element-selenium, vascular endothelial growth factor (VEGF) and soluble Fas (sFas) in leukemia patients, serum selenium concentration was measured by atomic spectrometry and levels of VEGF and sFas were simultaneously detected by enzyme linked immunosorbent assay (ELISA). Relationships of selenium, VEGF and sFas were analyzed by linear correlation. The results showed that serum selenium concentration in newly-diagnosed patients and relapsed patients was significantly lower than that in the control group (p < 0.05), especially in relapsed group. VEGF and sFas concentrations of refractory/relapsed group were significantly higher than those in the control group and the remission group (p < 0.05). But no significant difference was found between the remission group and the control group (p > 0.05). In leukemia patients, negative correlation was observed between selenium and VEGF (r = -0.529, p < 0.01), so did between selenium and sFas (r = -0.432, p < 0.01). Positive correlation was found between VEGF and sFas (r = 0.663, p < 0.01). It is concluded that the selenium, VEGF and sFas may take part in the occurrence of MDR in leukemia patients. Selenium has negative correlation with VEGF and sFas, which means that it may be used as an effective assistant agent to improve therapeutic effect of chemotherapy. However, further study in more cases is needed to reach a definite conclusion.

Anti-leukemia activity of T cells impacted by dendritic cells added with sodium selenite / 中国实验血液学杂志

The study was purposed to explore the quantity, morphology and immunophenotype of dendritic cells (DC) acquired by co-cultivated system with 3 types of cytokines and sodium selenite (Se) from peripheral blood mononuclear cells (PBMNCs), and to investigate the effects of Se on inducing the cytotoxic T lymphocyte (CTL) to get specific anti-leukemic activity in vitro by DC pulsed with K562 cell frozen-thawed antigen (antigen cell loading). PBMNCs isolated from healthy donors were cultured in RPMI 1640 medium contained 10% FBS supplied with 3 cytokines (rhGM-CSF, rhIL-4, TNF-alpha) for 4 days, DCs harvested were divided into 4 groups, DCI: DC alone; DCII: DC + Se (adding 0.5 micromol/L of Se); DCIII: DC + K562 (pulsed with lysed K562 cells); DCIV: DC + Se + K562. Morphology of DCs was observed under microscope at day 7. The CD1a, CD40, CD83, and CD86 were detected by FCM. Cytotoxicity of T cells induced by DC were measured with LDH release test at day 12. The level of IL-12 in supernatant of cultured DCs were determined with ELISA. The results indicated that at 7th day DC in 4 groups showed characteristic morphology, the colony numbers of 4 groups were all higher than those before cultivation. There were no obvious differences of morphology and colony counts between DCI group and DCII group. The colony numbers of DCIII group and DCIV group increased, as well as the ratio of suspended cells enhanced. The expressions of CD1a, CD40, CD83 and CD86 in 4 groups of DC were significantly higher than those in PBMNC group (p < 0.01), the expressions of CD1a and CD40 in 4 groups of DC did not display significant difference (p > 0.05), the expressions of CD83 and CD86 in both DCIII group and DCIV group were all higher than those in DCI group and DCII group (p < 0.01), but their expressions of CD83 and CD86 in DCI and DCII were not significantly different (p > 0.05), as well as those in DCIII group and DCIV group. With the ratio of 25:1 between E:T, killing rate of CTL on K562 cells in 4 DC groups were 15.3 +/- 2.3%, 26.3 +/- 3.7%, 28.2 +/- 4.5% and 36.2 +/- 3.7% respectively, all obviously higher than those of T cell group without being sensitized by DCs (5.9 +/- 2.4%) (p < 0.01), The CTL effect in DCIV group was the highest, which was higher than those in other 3 DC groups (p < 0.01); the effects in both DCII and DCIII group were also higher than that in DCI group (p < 0.01), but their difference between DCII and DCIII groups did not show significance (p > 0.05). The levels of IL-12 in supernatant of DCI, DCII, DCIII and DCIV groups were 257.0 +/- 64.2, 328.1 +/- 43.9, 323.0 +/- 53.5 and 353.9 +/- 46.2 pg/ml respectively, all significantly higher than that in supernatant of T cell alone group without being sensitized by DCs (35.27 +/- 27.1 pg/ml) (p < 0.01), The levels in DCII, DCIII and DCIV groups were all higher than that in DCI group (p < 0.01), but their levels between DCII, DCIII and DCIV groups were not of significant difference (p > 0.05). It is concluded that matured DCs can be successfully obtained from PBMNCs by a culture system contained rhGM-CSF, rhIL-4 and TNF-alpha with or without low-dose of Se (0.5 micromol/L) in vitro. Using K562 cell frozen-thawed antigen, DC express more adhesive molecules and co-stimulating molecules (CD83, CD86), and increase the secretion of IL-12, as well as the killing effects of CTL on special target cells. Low dose of Se did not showed effects on quantity and morphology of matured DC harvested, as well as their expression of mature phenotypes, it raised levels of IL-12 secreted by DCs, reaching the same level as using K562 cell frozen-thawed antigen, and it showed synergistic effect on induction of CTL with K562 cell frozen-thawed antigen.

Effects of Na(2)SeO(3) on expression of VEGF in K562/ADR cells / 中国实验血液学杂志

In order to investigate the effects of Na(2)SeO(3) on expression of VEGF in K562/ADR cells, K562 and K562/ADR cells were treated with Na(2)SeO(3) at dose of 5 and 10 micromol/L. The expressions of VEGF in K562 and K562/ADR cells were detected by ELISA before and at the different time point after treatment. The mutiplie of reversion of resistance was detected by MTT method. The results showed that Na(2)SeO(3) at dose of 10 micromol/L could increase the sensitivity of K562/ADR cell to adriamycin, the multiple of reversion was 3.48. The expression levels of VEGF in K562 and K562/ADR cells increased with prolongation of time cultured, and the VEGF expression levels in K562/ADR cells at the different time points were higher than that in K562 cells (P < 0.05); 5 and 10 micromol/L Na(2)SeO(3) did not suppress expression of VEGF in K562 cells at 72 hours (P > 0.05), and the VEGF level in K562 cells at 96 hours decreased without statistical significance; 5 and 10 micromol/L Na(2)SeO(3) acting for 48 hours did not show suppressive effect on expression of VEGF in K562/ADR cells (P > 0.05), 5 micromol/L Na(2)SeO(3) could decrease the expression of VEGF in K562/ADR cell after treatment for 96 hours, while 10 micromol/L Na(2)SeO(3) could significantly decrease the expression of VEGF in K562/ADR cells treated for 72 hours and 96 hours (P < 0.01). It is concluded that VEGF would be involved in the multidrug resistance of leukemia. Na(2)SeO(3) decreasing expression of VEGF in leukemic cells may be one of the mechanisms reversing multidrug resistance.

Analysis on the epidemiologic characteristics of human rabies in all the 31 provinces in China, from 1991 to 2005 / 中华流行病学杂志

<p><b>OBJECTIVE</b>To study the epidemic intensity and trend of human rabies from 1991 to 2005 among 31 provinces, metropoli and municipalities in China so as to increase the awareness of the disease.</p><p><b>METHODS</b>Contrastive analyses were performed and the annual publishing data by the Chinese Center for Disease Control and Prevention were used.</p><p><b>RESULTS</b>The total number of reported cases was 14 942 from 1991 to 2005 with an annual average mortality rate as 0.080/100000. The increase of five-years mortality ratio on relative ratio with circular base of mortality rate were--66.24% (1996-2000 to 1991-1995) and 506.13% (2001-2005 to 1996-2000). When comparing incidence rates between 2000-2005 and 1991-1995, the relative ratio with fixed base increase became 104.62%.</p><p><b>CONCLUSION</b>Among the 31 provinces, metropolis and municipalities, 27 had reported human rabies cases. The enzootic areas mainly distributed in the drainage area along the Yangtze River. The incidence rates of Guangxi, Hunan, Guizhou, Jiangxi and Guangdong were the highest.</p>

Virological surveillance on hemorrhagic fever with renal syndrome in Hunan province in 2006 / 中华流行病学杂志

<p><b>OBJECTIVE</b>To understand the rate of viral carrying status among rodents as well as genotypes and distribution of Hantaviruses (HV) isolated in Hunan province.</p><p><b>METHODS</b>With DFA, the HV antigen in lung tissues of rodents was detected. The total viral RNA was extracted from the lung tissues of the HV infected rats and amplified with reverse transcrition-polymerase chain reaction (RT-PCR), using the HV genotype specific primers. The amplified genes were then sequenced and subjected to genotyping and homologic analysis.</p><p><b>RESULTS</b>The average density of rodents was 3.15% and the virus carrying rate among rodents was 1.31%. Data from genotype analysis showed that the HV isolated from seven lung specimens taken from Rattus norvgicus, Apodemus agraius, Mus musculus, Rattus flavipectus among indoor rodents in Shaodong and Liuyang belonged to HV type II (SEOV), and one isolated from Apodemus agraius in Shaungfen belonged to HV type I (HTNV) among outdoor rodents. Six strains were sequenced successfully and the homology between six srains was 88.3%-100%. The homology of HN1, HN2, HN4, HN6 came from Liuyang and the HN7 and HN8 from Shaodong were both 100% while the homology between L99 and the strains from Liuyang and Shaodong were 94.4% and 88.3% respectively.</p><p><b>CONCLUSION</b>HV type II (SEOV) and the HV type I (HTNV) were all existed in Hunan province while SEOV was the main genotype.</p>

Reversal effect of sodium selenite on multidrug resistance in K562/ADR cell line and its mechanisms / 中国实验血液学杂志

This study was purposed to investigate the reversal effect of sodium selenite on multidrug resistance in adriamycin-resistant leukemic cell line K562/ADR and its mechanisms. The cytotoxicity and the reversal effect of sodium selenite on K562/ADR cells were assayed by MTT method; the apoptosis rate of K562 and K562/ADR cells were detected by flow cytometery, the mRNA expressions of mdr1 and bcl-2 were measured by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR). The results showed that 10 micromol/L sodium selenite significantly increased the cytotoxicity of adriamycin to K562/ADR cell and the reverse index (RI) was 2.31; the early apoptosis rate of K562 cells was elevated after treatment with 5 micromol/L Na(2)SeO(3) for 48 hours; and the medium-term and late apoptosis rate was elevated after treatment with both 5 and 10 micromol/L Na(2)SeO(3) for 48 and 72 hours. Both doses of 5 and 10 micromol/L Na(2)SeO(3) increased the early apoptosis rate of K562/ADR at 48 hours, and also increased the medium-term and late apoptosis rate after treating for 48 and 72 hours. The apoptosis rate was higher at dose of 10 micromol/L than that at 5 micromol/L, the apoptosis rate at 72 hours also was higher than that at 48 hours. The expressions of mdr1 mRNA and bcl-2 mRNA were decreased significantly by 10 micromol/L sodium selenite. It is concluded that sodium selenite can reverse the multidrug resistance in K562/ADR partially by down-regulating the expressions of mdr1 mRNA and bcl-2 mRNA, and increasing apoptosis rate of K562/ADR cells.

Effect of anti-TGF-beta antibody on ex vivo expansion and expression of adhesion molecules of human cord blood CD34+ cells / 中国实验血液学杂志

TGF-beta, as an inhibitor of hemopoiesis, excreted by hematopoietic stem and progenitor cells, down-regulates the expression of cytokines such as Flt-3 ligand, SCF, IL-3 etc on the stem and progenitor cells. The effect of anti-TGF-beta antibody on ex vivo expansion and expression of adhesive molecules on cord blood CD34(+) cells was studied in this research. The CD34(+) cells from six units of fresh umbilical cord blood were enriched by density gradient sedimentation and purified by miniMACS cell isolation system, and plated them into the SFEM serum free culture system which containing SCF, Flt-3L, TPO and IL-3 in the condition of 37 degrees C, 5% CO2, and saturated moisture. There were three groups in this experiment: (1) blank group: same as the culture system described above; (2) control group: added with normal rabbit IgG into the mentioned culture system; (3) test group: the same culture system with anti-TGF-beta1 antibo-dy. Cultured for 6 days, the number of mononuclear cells (MNC) was counted, the expression of CD34 antigen, CD117 (c-kit) antigen, CD11a antigen, CD49d antigen and CD33 antigen was tested with FCM. Meanwhile, cells of the three groups were plated in the methylcellulose culture system for 14 days, the number of CFU-GEMM, BFU-E, CFU-GM was counted. The results indicated that the expansion multiples of MNC, CD34(+) cells, CD34(+)c-kit(+) cells, CFU-GEMM in the test group (41.82 +/- 13.49, 15.62 +/- 6.95, 13.36 +/- 6.12, 11.07 +/- 4.05) were significantly higher than in the control group (28.86 +/- 9.03, 10.40 +/- 4.98, 9.04 +/- 4.40, 6.36 +/- 2.37) (P = 0.001, 0.002, 0.003, 0.002) respectively. The expansion multiple of more primitive CD34(+)c-kit(-) subpopulation in the test group (69.10 +/- 41.06) was even higher than in the control group (27.29 +/- 10.40) (P = 0.024). Adhesion molecule expression on the CD34(+) cells after short-term expansion: the expression of CD11a on the CD34(+) cells of the original cord blood was (61.73 +/- 4.13)%, and CD49d was (55.12 +/- 5.22)%. After expansion in each group the expression of CD11a on the CD34(+) cells did not change with statistical significance (P > 0.05), the expression of CD49d increased (P < 0.05). Compared with blank group and control group, anti-TGF-beta antibody did not impact on the expression of CD11a and CD49d (P > 0.05). It is concluded that anti-TGF-beta antibody can synergize other cytokines to effectively enhance the proliferation of cord blood NC, CD34(+) cells, progenitor subpopulation of CD34(+)c-kit(-) cells, and increase the output of more primitive progenitor colony, CFU-GEMM and BFU-E. At the same time, anti-TGF-beta antibody did not depresss the expression of adhesion molecules on CD34(+) cells.

The effect of all-trans retinoid acid and sodium selenite (Na2SeO3) on VEGF and its receptor expression in HL-60 cells / 中国实验血液学杂志

In order to investigate the effect of non-medullar toxicity drug - all trans retinoid acid (ATRA) and cancer preventive trace element-selenium compound - sodium selenite (Na(2)SeO(3)) on the expression of vascular endothelial growth factor (VEGF) and its receptor in HL-60 cells, the expression of VEGF and its receptor in HL-60 cells were detected by ELISA technique and flow cytometry before and after treatment with two drugs. The results showed that the mean VEGF concentrations in the cultural supernatant of 5 and 10 micro mol/L ATRA-treated HL-60 cells for 48 and 72 hours were lower than those of the control group without adding ATRA. The differences between the ATRA-treated groups and the control group were statistically significant (P = 0.001, P = 0.000, P < 0.01, respectively). The levels of VEGF-R on the surface of HL-60 cells also decreased after treatment with ATRA of 5 and 10 micro mol/L for 72 hours, but at 48 hours the expression rates of VEGF-R on HL-60 cells of the two ATRA treated groups were not significantly decreased. At 48 and 72 hours, Na(2)SeO(3) of 5 and 10 micro mol/L had no obvious effect on HL-60 secreting VEGF, but notablely inhibited the expression of VEGF-R. In conclusion, ATRA could inhibit the expression of VEGF and its receptor in HL-60 cell. Na(2)SeO(3) could not inhibit the expression of VEGF in HL-60 cell, but could decrease the receptor expression of VEGF, which mechanism should be further studied. ATRA and Na(2)SeO(3) had not obvious medullar-inhibition, but anti-angiogenesis activity. It is suggested that combination of two drugs with conventional therapy may enhance the effect of radiotherapy and chemotherapy, and reduce the dose and thus toxicity of chemotherapeutic agents.

The expression of vascular endothelial growth factor and its receptor in hematopoietic malignant cell lines HL-60 and Raji / 中国实验血液学杂志

In order to explore the effect of vascular endothelial growth factor (VEGF) in hematological malignancies, the expression of VEGF and its receptor was detected in HL-60 and Raji cells by reverse transcriptase-polymerase chain reaction (RT-PCR), enzyme linked immunosorbent assay (ELISA) and immunohistochemistry. The results showed that VEGF-mRNA expressed in both HL-60 and Raji cells, and the mean VEGF concentrations in the cultural supernatant of both cell lines were significantly higher than that of normal peripheral blood mononuclear cell respectively. There was expression of VEGF-R (Flt-1) on the surfaces of both HL-60 and Raji cells. The research results demonstrated that VEGF-mRNA was expressed in hematopoietic malignant cell lines (HL-60 and Raji), and the corresponding protein was secreted into the extracellular microenvironment, the both cell lines expressed VEGF-R on the cell surface. VEGF affects not only vascular endothelial cells, but also leukemic and lymphoma cells themselves. It is suggested that an autocrine pathway of VEGF existed in the both cell lines other than the paracrine pathway. The autocrine pathway of VEGF works as basis of tumor invasion. In conclusion, to restrain expression of VEGF and its receptor may inhibit tumor growth, and helps to block the reciprocal loop between VEGF and endothelial cells, and decrease the tumor specialities of hyperproliferation, anti-apoptosis and invation, that may make the tumor more susceptible to chemotherapy.

GPI-Pr Deficiency and Apoptosis of PNH Granulocytes / 中国实验血液学杂志

To study the relationship of Glycosyl phosphatidylinositol anchored proteins (GIP-Pr) and apoptosis of paroxysmal nocturnal hemoglobinuria (PNH) cells, we isolated peripheral granulocytes from 10 patients with PNH and 10 normal controls and measured apoptosis induced by serum starvation. The FCM analysis of phosphotidylserine (ps) externalization in granulocytes was determined using Annexin-V-FLUOS labeling. After the cells were induced for apoptosis in serum-free medium for 20 hours, the percentage of externalization was 78.6% in normal control cells but 39.5% in PNH cells. The results of FCM analysis of PI stained granulocytes showed that the PI positive rate was 51.5% in control cells and 30.2% in PNH cells. The gel electrophoresis analysis of DNA fragmentation all indicate that PNH granulocytes were relatively resistant to apoptosis as compared with normal controls. This resistance to apoptosis might not be related to the percentage of CD59 deficient granulocytes.