Prokaryotic expression, subcellular localization and enzymatic activity analysis of DXS gene from Platycodon grandiflorum / 药学学报

1-Deoxy-D-xylulose-5-phosphate synthase (DXS), the first key enzyme in 2-methyl-D-erythritol-4-phosphate (MEP) pathway, catalyzes the condensation of glyceraldehyde-3-phosphate with pyruvate to 1-deoxy-xylose-5-phosphate (DXP). In this study, PgDXS1, PgDXS2, and PgDXS3 genes were cloned from the root of Platycodon grandiflorum (P. grandiflorum). The open reading frame (ORF) of PgDXS1, PgDXS2, and PgDXS3 were 2 160, 2 208, and 2 151 bp in full length, encoding 719, 735, and 716 amino acids, respectively. Homologous alignment results showed a high identity of PgDXSs with DXS in Hevea brasiliensis, Datura stramonium and Stevia rebaudiana. The recombinant expression plasmids of pET-28a-PgDXSs were constructed and transformed into Escherichia coli (E. coli) BL21 (DE3) cells, and the induced proteins were successfully expressed. Subcellular localization results showed that PgDXS1 and PgDXS2 were mainly located in chloroplasts, and PgDXS3 was located in chloroplasts, nucleus and cytoplasm. The expression of three DXS genes in different tissues of two producing areas of P. grandiflorum were assayed via real-time fluorescence quantitative PCR, and the results showed that all of them were highly expressed in leaves of P. grandiflorum from Taihe. Under methyl jasmonate (MeJA) treatment, the expression levels of three PgDXS genes showed a trend of first decreasing and then increasing at different time points (3 - 48 h), and the activity of DXS showed a trend of first increasing and then decreasing in three tissues of P. grandiflorum. This study provides a reference for further elucidating the biological function of PgDXS in terpenoid synthesis pathway in P. grandiflorum.

Study on secondary metabolites of endophytic fungus Alternaria alternate from Paeonia lactiflora / 中草药

Objective This study focused on the secondary metabolites of endophytic fungus Alternaria alternate in Paeonia lactiflora. Methods Compounds were isolated from the EtOAc extract by chromatography technology and their structures were elucidated on the basis of comprehensive spectroscopic analysis. Results A total of 15 compounds were isolated and their structures were identified as methyl 4-acetamido-3-hydroxybenzoate (1), (E)-methyl5-hydroxy-3-methylpent-2-eno (2), isobenzofuranone A (3), 4-hydroxyacetophenone (4), 6-hydroxy-isosclerone (5), talarojlavone (6), 7-hydroxy-2-hydroxymethyl-5-methyl-4H-chromen-4-one (7), alternarienonic acid (8), 7-hydroxy-2,5-dimethyl-4H-1-benzopyran-4-one (9), 5-hydroxy-epialtenuene (10), alternariol (11), methyl 3-hydroxybenzoate (12), stemphyperylenol (13), altenusin (14), and altenuene (15). Conclusion Compounds 1, 3, 5-10, 12, and 13 are isolated from Alternaria alternatefor the first time.

Application of an IT-based follow-up platform in improving the outcome of patients with ischemic stroke / 中华医院管理杂志

Objective To establish an IT-based follow-up platform, and to explore its application effect in patients with ischemic stroke.Methods By constructing a follow-up model and a recurrence risk warning model for ischemic stroke patients, such a follow-up platform was established.Thanks to the retrospective comparative and analysis method, we built a study group comprising ischemic stroke patients discharged since the platform and a control group comprising 228 such patients discharged prior to the platform.These two groups were followed up by means of IT-based manner and traditional paper-based manner respectively at the first,third,sixth,ninth,and twelfth months since their discharge.These patients were analyzed in terms of their medication adherence,activities of daily living and recurrence rate.Results One year after the follow-up,32 cases were lost of contact in the study group and 42 cases from the control group.Medication adherence of the study group was higher than that of the control group at the sixth month (2.72 ±0.62), ninth month(2.86 ±0.37)and twelfth month(2.83 ±0.40)after discharge, with the differences being statistically significant(P <0.05).The recurrence rate of the study group at the ninth months(6.38%)and twelfth months(10.21%)after follow-up was lower than that of the control group,a difference being statistically significant(P<0.05).The difference of BI scores between the two groups was not statistically significant(P>0.05).Conclusions The IT-based follow-up platform could improve the medication adherence of ischemic stroke patients,and reduce the recurrence rate of ischemic stroke,but the effect of improving activities of daily living was still not significant.

Clinical Observation of Viper Bites Injury Limb Swelling Treated by Fushe Jiedu Decoction Combined with Red Light / 中国中医药信息杂志

Objective To explore the efficacy of Fushe Jiedu Decoction combined with red light on viper bites injury limb swelling and the effects on the inflammatory cytokines. Methods Totally 90 patients were divided into control group and experimental group by using random number table method, with 45 cases in each group. The wounds of the control group were sterilized and given anti-snake venom serum, antibiotics, and tetanus immunoglobulin and supplemented with energy to correct water and electrolyte disturbances. The experimental group was treated with Fushe Jiedu Decoction based on the treatment of control group,150 mL each time,orally,twice a day; Red light was applied at the site of the most obvious swelling of the injured limb, 20 minutes each time, twice a day. The treatment lasted for 6 d. The swelling of injured limbs, serum C-reactive protein (CRP), 5-hydroxytryptamine (5-HT), histamine, tumor necrosis factor-α (TNF-α), interleukin (IL)-6 levels before and after treatment in the two groups were observed. Results Compared with before treatment, the swelling of the limbs disappeared significantly in the experimental group at 3 d and 6 d and in the control group at 6 d, with statistical significance (P<0.01). Compared with the control group at the same time point, the swelling of the limbs in the treatment group was significantly better than that in the control group at 3 d and 6 d, with statistical significance (P<0.01). Compared with before treatment, the levels of CRP, 5-HT, TNF-α, and IL-6 were significantly lower in the two groups, with statistical significance (P<0.05). After treatment, CRP, 5-HT and histamine in the experimental group were significantly better than that in the control group(P<0.05).Conclusion Fushe Jiedu Decoction combined with red light has good efficacy for viper bites injury limb swelling, which can reduce inflammatory cytokines levels of patients.

Application of microsutures with vascular endothelial growth factor to improve vascular endothelial regeneration after small vessel anastomosis in the rats / 中国组织工程研究

BACKGROUND:The vascular endothelial growth factor (VEGF) plays an important role in the development and formation of blood vessels.Up to now,there are few reports about the treatment of postoperative complications of vascular anastomosis surgery by mcrosutures with VEGF in China.OBJECTIVE:To synthesiize microsutures with VEGF and to evaluate its effect in revascularization following small vessel anastomosis.METHODS:The method of emulsification-diffusion was use to produce biodegradable polymer polylactic acid/glycolic acid (PLGA) copolymer microparticles containing VEGF,and then,the microparticles were added into microsutures to prepare microsutures with VEGF.Ninety Sprague-Dawley rats were enrolled to make animal models of caudal artery anastomosis using microsutures with VEGF in experimental group and microsutures alone in control group.Complications and VEGF level in the peripheral blood were detected and hematoxylin-eosin staining at the anastomotic site was performed at 2,12 hours,1,3,7 days after anastomosis.RESULTS AND CONCLUSION:(1) Postoperative complications:The postoperative incidence of skin necrosis was significantly lower in the experimental group than the control group (P ＜ 0.05).(2) VEGF level:Compared with the control group,the peripheral blood VEGF level was significantly higher in the experimental group at each time point after operation (P ＜ 0.05).(3) Hematoxylin-eosin staining:In the experimental group,proliferated endothelial cells were seen near the anastomotic site at 1 day after anastomosis;there were a large number of proliferated endothelial calls and subcutaneous tissues covering the sutures completely at 3 days after anastomosis;and endothelial cells and internal elastic lamina were completely repaired,smooth muscle cells proliferated further,and the outer membrane returned to normal at 1 week after anastomosis.In the control group,cell degeneration and necrosis were seen near the anastomotic suture,and only adventitial cells infiltrated and exhibited a traumatic proliferative response at 1 day after anastomosis;neonatal endothelial cells appeared in the exfoliated area of the endothelial cells,grew and migrated,and there was a few endothelial cells covering the anastomotic site at 3 days after anastomosis;and newborn endothelial cells got over the anastomotic crack and covered the suture.To conclude,microsutures with sustained-release VEGF microparticles can promote endothelial cell regeneration in rats at the anastomotic site.

Secondary metabolites from endophytic fungus Fusarium oxysporum isolated from Paeonia ostii / 中草药

Objective To study the secondary metabolites of endophytic fungus Fusarium oxysporum form Paeonia ostii. Methods The fermentation liquor of the fungal strain F. oxysporum were isolated and purified using various chromatographic methods. The structures of the compounds were identified by spectrum analysis. Results Twenty-three compounds were isolated from the fungal and their structures were identified as (2S)-proline (1), (22E,24R)-ergosta-7,22-diene-3β,5α,6β-triol (2), cyclo-(S-Pro-S-Leu) (3), cyclo-(L-Ala-L-Pro) (4), cyclo-(Val-Pro) (5), cyclo-(R-Pro-S-Phe) (6), cyclo-(D-cis-Hyp-L-Phe) (7), cyclo-(trans-4-hydroxy- L-Pro-L-Phe) (8), cyclo-(Gla-Tyr) (9), cyclo-(trans-4-hydroxy-L-Pro-L-Leu) (10), L,L-cyclo-(Trp-Pro) (11), cyclo-(L-Leu-Gly) (12), N-(3-(1H-indol-3-yl)propyl)acetamide (13), indole-3-acetic acid (14), 2-piperidone (15), 2-pyrrolidinone (16), thymidine (17), (5S)-5-[(4-hydroxyphenyl)methyl]-2,4-imidazolidinedione (18), (3S,6S)-3-(butan-2-yl)-6-(1-hydroxyethyl) piperazine-2,5-dione (19), cyclo-(Ala-Leu) (20), cyclo-(S-Pro-S-Phe) (21), cyclo-(Tyr-Pro) (22), and cyclo-(L-Phe-L-Tyr) (23). Conclusion With the exception of compounds 2, 6, and 14, the other compounds are isolated from the fermentation liquor of the fungal strain F. oxysporum for the first time.

Effect of total flavonoids in Scutellaria barbata in mediating autophagy in tumor cells via PI3K/AKT/mTOR pathway / 中国中药杂志

To investigate the effect of the total flavonoids in Scutellaria barbata(TF-SB) against autophagy in tumor cells in vivo, and further determine whether the mechanism is correlated with the PI3K/AKT/mTOR pathway, which lead to autophagy-induced tumor cell death. Melanoma-bearing mice were prepared and divided into control group, rapamycin group (Rap 1.5 mg•kg⁻¹), and high, middle and low-dose TF-SB (200, 100, 50 mg•kg⁻¹) groups. The groups were given drugs once a day for successively 11 days. The inhibitory effect of TF-SB on the growth of melanoma was determined by measuring tumor volume and tumor inhibition rate. TUNEL method was used to detect the apoptosis of tumor cells to further verify the antitumor activity of TF-SB. The protein expressions of LC3-Ⅰ and LC3-Ⅱ were detected by Western blot, and the relative expression of LC3-Ⅱ was calculated based on LC3-Ⅱ/LC3-Ⅰ. In addition, the effect of TF-SB on autophagy of tumor cells, the underlying molecular mechanism of TF-SB in inducing autophagy and PI3K/AKT/mTOR pathway marker protein phosphorylation were also studied. According to the results, TF-SB effectively inhibited melanoma growth in mice, reduced tumor volume, increased the tumor inhibition rate, and significantly increased tumor cell apoptosis index and the ratio of LC3-Ⅱ/LC3-I (P<0.05, P<0.01 or P<0.001). The protein expressions of p-PI3K, p-AKT and p-mTOR were also suppressed dramatically compared with those in control group (P<0.05, P<0.01 or P<0.001). In conclusion, the total flavonoids in S. barbata could inhibit the growth of melanoma in vivo by inducing autophagy and apoptosis of tumor cells, which may be correlated with suppression of PI3K/AKT/mTOR pathway.

Total flavonoids in Scutellaria barbata prevents NLRP3 inflammasome expression in tumor cells by affecting autologous pathway / 中国中药杂志

This paper was aimed to investigate the relationship between autophagy and NLRP3 inflammasome activation by studying the effect oftotal flavonoids in Scutellaria barbata (TF-SB) on autophagy in tumor cells and NLRP3 inflammasome, and to provide experimental evidence for further study of the anti-tumor mechanism of TF-SB. Mielanoma models were established by inoculating B16-F1 cell line to mice, and then were randomly divided into 5 groups (n=10 in each group): model control, positive control control(Rap, 1.5 mg•kg⁻¹), and TF-SB low, middle and high groups (50, 100 and 200 mg•kg⁻¹). Meanwhile, healthy C57BL/6J mice were used as normal control group (n=10). The drugs were given once daily for 2 weeks consecutively. Thirty minutes after last treatment, the determinations at endpoint were performed; pathological changes of tumor tissue were evaluated by using HE staining; protein expressions of LC3-II/LC3-I or NLRP3inflammasome/caspase-1/IL-1β and IL-18 in tumor tissues were detected by using Western-blot; and serum levels of IL-1β and IL-18 were detected by using Elisa kit. The results showed that the tumor cells in model group showed obvious atypia and malignant proliferation; the invasion of tumor tissue was significantly reduced, the tumor necrosis area was significantly increased, and the inflammatory reaction was significantly alleviated in positive control group and various TF-SB groups. As compared with model control group, LC3-II/LC3-I was significantly increased, while NLRP3/caspase-1/IL-1βand IL-18 protein expressions were significantly decreased in positive control group and TF-SB groups. Serum IL-1β and IL-18 levels in model control group were found higher than those in control group (P<0.001), but they were significantly lowered in positive control group and TF-SB groups (P<0.05, P<0.01 or P<0.001). Taken together, total flavonoids in S. barbata could effectively alter the tumor growth micro-environment by inhibiting the expression of NLRP3 inflammasome, and its anti-tumor effect may be associated with the induction of tumor cell autophagy.

Protective effects of Sapindus saponins in spontaneously hypertensive rats / 中国结合医学杂志

<p><b>OBJECTIVES</b>To investigate the protective effects of Sapindus saponins in spontaneously hypertensive rats, and the possible cellular and molecular mechanisms.</p><p><b>METHODS</b>Thirty-two 16-week-old spontaneously hypertensive rats were randomly divided into four groups (8 in each group): model group (placebo), positive control group (27 mg/kg of Captopril Tablets), Sapindus saponins groups (27 mg/kg and 108 mg/kg, respectively). Another 8 healthy Wistar-Kyoto strain (WKY) rats were used as the normal group. The animals were treated for 8 weeks. Blood pressure of rats was determined by non-invasive blood pressure meter (BP-6). Furthermore, the contents of angiotensin II (Ang II) in plasma and myocardial tissue were determined by enzyme-linked immunosorbent assay (ELISA), the gene expression of receptor angiotensin type 1 (AT1R) in aorta was determined by quantitative realtime polymerase chain reaction (qRT-PCR). The protein expression of transforming growth factor-β1 (TGF-β1) and AT1R in heart was determined by immunohistochemical staining. The protein expression of p-phosphorylation of p38 mitogen-activated protein kinase (p-p38MAPK) was determined by Western blotting. The contents of interleukin (IL)-1, IL-6 and tumor necrosis factor (TNF) in serum were determined by radioimmunoassay. And the histopathological and morphological changes of aorta and heart tissue samples were assessed semi-quantitatively by hematoxylin-eosin (HE) or Masson staining.</p><p><b>RESULTS</b>Thirty minutes after single or continuous treatment, systolic blood pressure (SBP) was reduced significantly in Sapindus saponins groups. And the contents of AngII, IL-1, IL-6 and TNF-α in serum, the expression of AT1R mRNA, p-p38MAPK and TGF-β1 were significantly suppressed dose-dependently (P<0.05 or P<0.01). With the Sapindus saponins treatment, compared with those of the model group, the cardiac and aortic pathological changes were ameliorated significantly.</p><p><b>CONCLUSIONS</b>Our findings suggest that Sapindus saponins might have protective effects in spontaneously hypertensive rats, the cellular and molecular mechanisms of which might be relevant to the regulation of inflammatory responses mediated by p-p38MAPK signal pathway based on activated Ang II and AT1R.</p>

Therapeutic strategies for a functional cure of chronic hepatitis B virus infection

Treatment of chronic hepatitis B virus (HBV) infection with the viral DNA polymerase inhibitors or pegylated alpha-interferon has led to a significant retardation in HBV-related disease progression and reduction in mortality related to chronic hepatitis B associated liver decompensation and hepatocellular carcinoma. However, chronic HBV infection remains not cured. The reasons for the failure to eradicate HBV infection by long-term antiviral therapy are not completely understood. However, clinical studies suggest that the intrinsic stability of the nuclear form of viral genome, the covalently closed circular (ccc) DNA, sustained low level viral replication under antiviral therapy and homeostatic proliferation of hepatocytes are the critical virological and pathophysiological factors that affect the persistence and therapeutic outcomes of HBV infection. More importantly, despite potent suppression of HBV replication in livers of the treated patients, the dysfunction of HBV-specific antiviral immunity persists. The inability of the immune system to recognize cells harboring HBV infection and to cure or eliminate cells actively producing virus is the biggest challenge to finding a cure. Unraveling the complex virus-host interactions that lead to persistent infection should facilitate the rational design of antivirals and immunotherapeutics to cure chronic HBV infection.

Protection effects of S-adenosyl-L-methionine on lead-exposed rats during development and its mechanism of long-term potentiation / 中华预防医学杂志

<p><b>OBJECTIVE</b>To explore the effects of S-adenosyl-L-methionine (SAM) on blood lead concentration and oxidative stress of tissue in prenatal and postnatal lead-exposed rats, and evaluate the potential reparation exerted by SAM on paired-pulse facilitation (PPF) and long-term potentiation (LTP) in lead-exposed rat.</p><p><b>METHODS</b>Pregnant Wistar rats were randomly divided into three groups: control, lead-exposed and lead-exposed with SAM treatment groups. Lead-exposed rats drank 1.5 g/L lead acetate solution through pregnancy until weaning and then the pups received 20 mg/kg SAM or saline daily intraperitoneally depending on their group. Control group rats drank tap water throughout the experiment. At the postnatal 44-60 days, all the pup rats were given an extracellular recording measured in dentate gyrus (DG) area of hippocampus. The blood lead concentration and oxidative stress in liver, brain and hippocampus were also detected.</p><p><b>RESULTS</b>The blood lead concentration in lead-exposed group was higher (159. 3 +/- 10. 9 microg/L) in comparing with those of control group (27.5 +/-3.8 microg/L) and lead +SAM group (33.1 +/-9.5 microg/L) (F=213.5, P<0.01). A significant recovery of liver, brain glutathione (GSH) and malondialdehyde (MDA) level was clearly produced in lead-exposed rats after SAM treatment (P <0.05). Chronic lead exposure during development impaired LTP measured on field excitatory postsynaptic potential (EPSP) [(112 +/-2.1)%] compared with control rats [(131+/-4.5)%] and the impaired LTP could be significantly increased by SAM treatment [(120 +/- 2.6)%] (F = 26. 1, P <0. 05).</p><p><b>CONCLUSION</b>SAM might be beneficial for treatment of lead intoxication, especially in the rescue of learning and memory impairment induced by lead and should deserve more detailed research.</p>

Isolation and identification of arboviruses in Hebei Province / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To study the arboviruses carried by mosquitoes collected in Hebei Province.</p><p><b>METHODS</b>Samples were collected from mosquito active sites and stored in liquid nitrogen till use. Pools of 20 to 30 mosquitoes were ground after sterilization, centrifugal supernant was inoculated onto C6/36 cell, cytopathic effect was observed for three sequential passages. Positive isolates were identified by IFA and RT-PCR.</p><p><b>RESULTS</b>Totally 1310 mosquitoes were collected from two villages of She county, Hebei province. They were divided into 46 pools and ground respectively. Thirteen positive isolates were obtained. Two isolates reacted with alphaviral antibodies and were amplified by alphaviral primers, nucleotide sequence showed the highest homology (98%) to Getah virus (AY702913.1), so the two isolates were identified as Getah virus.</p><p><b>CONCLUSION</b>Getah virus was isolated from mosquitoes in Hebei Province. This is the first report of isolating Getah virus from inland of China.</p>

Molecular biological identification of Batai virus isolated in China / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To study the molecular characteristics of YN92-4 strain isolated from mosquitoes in Yunnan Province and define its classification.</p><p><b>METHODS</b>The S segment of YN92-4 strain was amplified and sequenced by 2 different sets of primers. The phylogenic tree of S fragment was constructed by Phylip bio-software. The amino acid sequences of N and NSs proteins were also studied.</p><p><b>RESULTS</b>YN92-4 strain could be amplified by 2 sets of primers respectively, S segment showed a highest homology with Batai virus (X73464), reached 96.4%, the homology of protein N and NSs amio-acid sequence with Batai virus was 99.1% and 98% respectively.</p><p><b>CONCLUSION</b>The YN92-4 strain belongs to Batai virus, this is the first report of molecular biological identification of Batai virus in China.</p>

Proteomic analysis on metastasis-associated proteins of hepatocellular carcinoma tissues / 中华肝脏病杂志

<p><b>OBJECTIVE</b>A comparative proteomic approach was used to identify and analyze proteins relevant to metastasis of hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>Proteins extracted from 12 liver tumor tissue specimens (6 with metastases and 6 without) were separated by two-dimensional gel electrophoresis (2-DE). Comparative analyses of 2-DE protein patterns between the two groups were done using computerized image analysis. Selected proteins exhibiting statistically significant alternations were identified by mass spectrometry. Immunohistochemistry, Western blotting and RT-PCR were performed to examine the expressions of the candidate proteins.</p><p><b>RESULTS</b>16 proteins including HSP27, S100A11, CK18 were identified using mass spectrometry, which were related to cell mobility, signal transduction, and energy metabolism respectively. Of these, HSP27 was found to be uniquely over-expressed in 2-DE maps of all metastatic HCCs when compared to the non-metastatic HCC tissues. Immunohistochemistry and Western blotting of HCC tissues confirmed this difference while RT-PCR did not.</p><p><b>CONCLUSION</b>There are different proteins working together that affect the metastasis of HCCs. The overexpression of HSP27 may serve as a biomarker for early detection and therapeutic targets to the metastatic phenotype of HCC. The role of HSP27 in HCC metastasis warrants further investigation.</p>

Differential analysis of tyrosine-phosphorylated proteins in human hepatocellular carcinoma cell lines with different metastasis potentials / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To compare expressions of tyrosine-phosphorylated proteins in different hepatocellular carcinoma cell lines with different metastasis potential and to screen key molecules associated with HCC metastasis and recurrence.</p><p><b>METHODS</b>Using two-dimensional electrophoresis, Western blotting and MALDI-TOF-MS/MS, we analyzed tyrosine-phosphorylated protein profiles of Hep3B, MHCC97L and MHCC97H, HCC cell lines with different metastasis potentials.</p><p><b>RESULTS</b>10 spots were detected in Hep3B, 19 in MHCC97L and 17 in MHCC97H. Seventeen significantly different phosphotyrosine proteins in gel were identified by MALDI-TOF-MS/MS, including Annexin I.</p><p><b>CONCLUSION</b>The changed expression of tyrosine-phosphorylated proteins is associated with HCC metastasis and recurrence.</p>

Studies of coltivirus in China / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>The purpose of this article is to review the developments of studies of Coltivirus in China.</p><p><b>DATA SOURCES</b>The data used in this review was obtained mainly from the studies of Coltivirus reported from 1990 to 2003 in China.</p><p><b>STUDY SELECTION</b>Relevant articles on studies of Coltivirus in domestic and foreign literature were selected.</p><p><b>DATA EXTRACTION</b>Data were maily extracted from the articles which are listed in the reference section of this review.</p><p><b>RESULTS</b>Many Coltiviruses have been isolated not only from blood samples of patients with unknown fever or from cerebrospinal fluid of patients with encephalitis in Xishuangbanna area in Yunnan province, but also from mosquitoes collected in many areas in China. In some patients diagnosed as Japanese encephalitis or unknown fever, an increase of Coltivirus IgG antibody of fourfold, or more, has been detected using ELISA. Similarly, Coltivirus IgM antibody was positive in some patients with Japanese encephalitis or viral encephalitis. From most Chinese patients, except the northeastern, the isolates of Coltiviruses belong to subgroup B2, according to RT-PCR amplification of the ninth and twelfth segments of the isolates and sequence analysis of their amplicons. Some biological properties of Chinese Coltiviruses isolates are different from that of North American Coltiviruses.</p><p><b>CONCLUSIONS</b>The isolates of Coltiviruses from Chinese patients are one of the common agents causing viral encephalitis and unknown fever in summer-autumn season. It might be an important public health problem due to its high isolation rate and wide distribution in China. Mosquito is the main transmission vector of the virus.</p>

Screening hepatocellular carcinoma autoantibodies by serological proteome analysis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To screen hepatocellular carcinoma (HCC) autoantibodies as diagnostic biomarkers or therapy targets by serologic proteome analysis (SERPA).</p><p><b>METHODS</b>Total proteins extracted from human HCC cell line HCCLM3 were separated by two-dimensional electrophoresis (2-DE) and then transferred onto PVDF membranes, which were subsequently incubated with sera from HCC, hepatitis B virus (HBV) infected patients or healthy volunteers. All immuno-reactive protein spots on blot films were matched to those on 2-DE gel maps by image analysis and identified by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS/MS).</p><p><b>RESULTS</b>2-DE gel maps of HCCLM3 and corresponding blot films of good quality and reproducibility were established. The number of spots on HCCLM3 2-DE reference gel totaled 603 and those on HCC, HBV and healthy sera blotted films were 70.75+/-24.25, 68.5+/-23.44 and 41.38+/-15.05, respectively. Blot films of HCC and HBV groups had more spots than those of the healthy group (P < 0.05) while no significance was found between films of HCC and HBV groups. By identification, those HCC autoantibodies could be classified as nuclear proteins, cytoskeleton proteins, heat shock proteins and metabolic enzymes.</p><p><b>CONCLUSION</b>Serological proteome analysis is a high throughput technique for screening tumor autoantibodies. Those newly identified HCC associated tumor antigens and corresponding autoantibodies can be used in the early diagnosis or immuno-therapy of HCC.</p>

Investigation of Arboviruses in Lancang river down-stream area in Yunnan province / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the epidemic state of arboviruses in the downstream area of Lancang river in Yunnan province.</p><p><b>METHODS</b>Mosquitoes were collected from Lancang river downstream area (including Lancang county and Simao city) during summer in 1998 and stored in liquid nitrogen after classification. The mosquito pools were homogenized and centrifuged, then the supernatant was inoculated into C6/36 cells for virus isolation. New isolates were identified by neutralization test(NT), ELISA, immunofluorescence assay(IFA) and polyacrylamid gel electrophoresis(PAGE).</p><p><b>RESULTS</b>Totally 22 isolates of arbovirus were obtained from 233 mosquito pools by inoculation of C6/36 cells and positive rate of the isolation was 9.4%. Ten strains were resistant to both ether and 5 prime-IDU. So they were non-enveloped double-stranded (ds) RNA virus. Twelve segmented RNAs were shown by PAGE and PAGE profiles from the ten strains were 6-6 with minor variation. The isolates can be neutralized by immunized mouse ascites fluid of BJ95-75 strains of coltivirus by NT, and reacted with monoclonal antibody against BJ95-75 by ELISA. These viruses were identified as coltivirus. Nine isolates were sensitive to ether and resistant to 5 prime-IDU. So they were non-enveloped RNA viruses. PAGE showed 10 segmented RNA, and they were identified to be orbiviruses. Three isolations were sensitive to ether. One of them can be neutralized with JEV A2 strain antibody by NT and was positive to the homologous antibody by IFA. It was identified being strain of JE virus. One strain(YN92-4) can be reacted with anti-bunyavirus group specific immune ascites fluid by both IFA and ElISA, but reacted neither with anti-alpha virus group, nor with anti-flavivirus group JE virus ascites fluid. The virions are spherical and about 87 nm in diameter with surface projections by negative staining. Conclusion Twenty-two isolates have been obtained from wild caught-mosquitoes of Lancang river down-stream area in Yunnan province. Among them ten, nine, one and one were identified as coltivirus, orbivirus, JE virus and bunyavirus, respectively. One is under identification. This is the first report on bunyavirus isolated from mosquitoes in China.</p>

Genotyping of the Chinese isolates of coltivirus / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To classify the Chinese isolates of Coltiviruses.</p><p><b>METHODS</b>Three sets of primers were selected among them two were specific to the 9th and 12th segments of subgroup B2, and one was for the 12th segment of subgroup B1-All the Chinese isolates of Coltivirus selected in the experiment were classified according to the lengths of different amplicons of the reverse transcriptase-polymerase Chain reaction (RT-PCR). The homogenicity of the nucleic acids of the isolates BJ95-75 and YN-6 was also compared with other Coltivirus strains belonging to subgroup B2.</p><p><b>RESULTS</b>With the primers 12-854-S/12-B2-R, which were specific to the 12th segment of Coltivirus subgroup B2-850 bp amplicons were obtained from Beijing isolate BJ95-75 and all the Yunnan isolates such as YN-6, -67-1, -68-1, -69, -70-1, -70-2, -90, -92-2, -93 of Coltivirus 492 bp DNA fragments were also amplified from all of them with the segment 9th specific primers 9-JKT-S/9-JKT-R. However no positive results were obtained from Northeast isolates NE97-12, NE97-31 and control viruses YN-99(Orbivirus),YN-151-1(JEV) with the same two sets of primers. With 12-B1-S/12-B1R primers specific to the 12th segment of subgroup B1, no amplicons of right length were obtained from any of the Chinese isolates of Coltivirus and the control viruses. When compared the nucleic acid sequences of BJ95-75 and YN-6 with other Coltivirus strains such as Bannavirus, JKT6423, JKT6969, JKT7043, the amplicons from segment 12th of these two strains had more than 89.4% homology with the other strains, especially to the earlier Chinese isolate Bannavirus, the homolog was more then 98.9%. Nearly 96.5% and 99.2% of the nucleic acids of the amplicons from segment 9th of the two strains were being homologous to Bannavirus and about 84.0% to JKT6423, which had been classified into type B2a. But the maximal homogenicity was about 53% when compared with the other two coltivirus strains. JKT6969 and JKT7043 which had been classified into type B2b.</p><p><b>CONCLUSION</b>Genotyping the recent Chinese isolates of coltivirus for the first time in our country. Most of the Chinese isolates belong to subgroup B2, more exactly type B2a. The Northeast isolates NE97-12 and NE97-31 were not correctly grouped with the available primers.</p>

Alteration of serum lipids in patients with acute leukemia and its clinical significance / 中国实验血液学杂志

To explore the clinical significance on alteration of serum lipids in acute leukemia (AL) patients, the level of serum lipids was monitored in 86 AL cases by using of automatic biochemical analyzer. The results showed that TG was significantly higher (P < 0.05) while TC, LDL-C and HDL-C were obviously lower (P < 0.05) in AL patients than those in normal controls. After chemotherapy, TG decreased but TC, LDL-C and HDL-C were still higher (P < 0.05) as comparing to pre-treatment in complete remission cases. There were no changes of those parameters in non-remission patients. It is concluded that determination of serum lipids level in AL patients is a simple and important accessory index to evaluate curative effect and monitor patient's condition.