AlM: To silent hypoxia inducible factor-1α ( HlF-1α) gene in malignant melanoma of the choroid cell by small interference RNA ( siRNA ) and investigate its effect on the expression of matrix metalloproteinase-2 ( MMP-2 ) in the choroid cell line human uveal melanoma cell (OCM-1) in hypoxia environment.METHODS:OCM-1 cells cultured on culture flask were divided into normal group and hypoxia group. Hypoxia group were divided into five groups: simple hypoxic group, and interference group, and negative control group, and positive control group, and liposome group. Normal group cells were cultured on DMEM culture flask with 10% FBS, 100U/mL penicillin, 100μg/mL streptomycin as well as high concentration of glucose. The cells were maintained at 37℃ in a humidified 5% CO2 incubator. Cells in good condition were selected for experiment. For hypoxia group, chemical hypoxia inducer CoCl2 was added into nutrient medium at the concentration of 100μmol/L to simulate hypoxia microenvironment. We designed and synthesised siRNA ( siRNA + negative control+positive control ) , the target sequences of the HlF-1α to transfect hypoxic malignant melanoma of the choroid cell. SiRNA including HlF-1α siRNA, β-actin siRNA and negative control group synthesized in vitro transfected hypoxic OCM - 1 cell through Lipofectamine2000. The expression of HlF-1α, MMP-2 gene and the protein were detected by RT-PCR and Western blot. RESULTS: Compared with the normal group, the expression of HlF-1α mRNA was not obviously changed (P>0. 05), but the expression of HlF-1α protein and MMP- 2 mRNA protein was significantly higher ( P<0. 05) . Compared with the other hypoxia groups,β-actin mRNA expression of positive control group decreased ( P< 0. 05 ) , which proved successful transfection. The expression of HlF-1α mRNA and the expression of its protein and both MMP-2 mRNA and its protein was significantly lower ( P < 0. 05 ). The negative control group, liposome control group had no significant difference in the detection of factors (P>0. 05).CONCLUSlON: Hypoxia status may upregulate the HlF-1α in OCM-1 cells by increasing the expression of protein. Hypoxia can also inactivate MMP-2, resulting in upregulation of MMP-2 RNA and the expression of its protein. The expression of HlF-1α and MMP-2 mRNA can be down-upregulated by transfecting OCM-1 with HlF-1α siRNA.

The seeds of Rabdosia rubescens were as the materials to research the impacts of different lead (Pb2+) concentrations(0, 135, 270, 540, 1 080 mg x L(-1)) on seed germination and seedling growth. The results show that: Low concentration of lead had no obvious effect on early germination of the seed, the germination vigor and germination speed were lightly higher but not significantly differed at the level of Pb concentration 135 mg x L(-1) with control group; Mid-high concentration of Pb solution (270-1 080 mg x L(-1)) significantly inhibited the seed germination and seedling growth, which reduced the seed germination rate, germination vigor, germination index, embryo root length and shoot length, growth index with increasing of Pb concentrations. There was a inhibitory effect on embryo shoot length and root length at mid-high lead concentrations stress, and stronger inhibitory effect on root , which was more sensitive than shoot to Pb stress(P < 0.05). Pb bioaccumulation coefficient (BC) was 0.76-2.59, increased with concentration of Pb; Pb enrichment in seedling mainly caused the growth inhibition. The fitting model predictive analyses show, the critical concentration of Pb, which causes the germination rate and biomass fresh weight reducing 10%, is 195.18, 101.65 mg x L(-1).
Germination ; drug effects ; Isodon ; drug effects ; growth & development ; Lead ; toxicity ; Seedlings ; growth & development ; Seeds ; growth & development ; Stress, Physiological

This study was aimed to investigate the T cell (helper T cells) immune status in ITP patients and its relation with therapeutic response. 20 de novo ITP patients were enrolled (8 males, 12 females) with a median age of 41 (20 to 81). Real-time RT-PCR method was used to measure the gene expression of Th cells including T-bet, IFN-γ, GATA-3, TGF-β, Foxp3, IL-2, IL-4 in PBMNC of ITP patients before and after conventional dose of prednisone therapy [1 mg/(kg·d)] and in PBMNC of 20 normal controls. The results showed that T-bet, IFN-γ and IL-2 were significantly over-expressed in PBMNC of ITP patients before treatment compared with that in normal controls (p < 0.01), and compared with that before treatment, T-bet, IFN-γ, and IL-2 were markedly down-regulated in ITP patients after treatment. Before treatment, the expressions of Foxp3, TGF-β, GATA3 and IL-4 in ITP patients did not show difference from normal controls, while after treatment Foxp3 were more up-regulated than that before treatment (p < 0.05). After treatment, TGF-β expression showed a different pattern between old and young patients. TGF-β expression was down-regulated (p < 0.05) among ITP patients younger than 60, while up-regulated in older patients. It is concluded that there is an imbalance of Th1/Th2/Treg cytokines in ITP patients, which can be reversed by glucocorticoid treatment. The conventional dose of glucocorticoid may be regarded as effective therapy for de novo ITP patients, it may correlate with improvement of imbalance between Th1/The2/Treg cytokines.
Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Female ; Forkhead Transcription Factors ; metabolism ; Gene Expression ; Gene Expression Regulation ; Glucocorticoids ; therapeutic use ; Humans ; Interferon-gamma ; metabolism ; Interleukin-2 ; metabolism ; Male ; Middle Aged ; Purpura, Thrombocytopenic, Idiopathic ; drug therapy ; metabolism ; pathology ; T-Box Domain Proteins ; metabolism ; T-Lymphocytes, Helper-Inducer ; metabolism ; Transforming Growth Factor beta ; metabolism ; Young Adult

Objective: To observe the clinical efficacy of Zhen'ai needling method in Nei Jing (Classic of Internal Medicine) for children with allergic rhinitis (AR) accompanied by adenoid hypertrophy (AH). Methods: A total of 74 children who met the screening criteria were divided into a Zhen'ai group and a control group by the random number table method, with 37 cases in each group. The control group was treated with acupuncture at Zusanli (ST 36), Hegu (LI 4), Yingxiang (LI 20), Juliao (ST 3), Yintang (GV 29), Shangxing (GV 23) and Baihui (GV 20). The Zhen'ai group added points of Zhen'ai needling method {Shanglianquan [Extra, located at 1 cun above Lianquan (CV 23)], Tianrong (SI 17) and Lieque (LU 7)} in addition to the points in the control group. The needles were retained for 30 min. The treatment was performed twice a week. The total nasal symptom score (TNSS), sino-nasal outcome test-20 (SNOT-20) and symptom scale for AH (SSAH) were assessed before and after 10 treatments. The clinical efficacy of the two groups was compared after treatment. Results: During the treatment, 5 cases dropped out in the control group and 2 cases in the Zhen'ai group. After treatment, the total effective rate of the Zhen'ai group was 94.3％, versus 93.8％ in the control group. There was no significant difference between the two groups (P>0.05). The markedly effective rate of the Zhen'ai group was 42.9％, versus 12.5％ in the control group, and the difference was statistically significant (P<0.05). After treatment, there were significant intra-group differences in the scores of TNSS, SNOT-20 and SSAH in both groups (all P<0.05); the scores of SNOT-20 and SSAH in the Zhen'ai group were lower than those in the control group, and the differences between the groups were statistically significant (both P<0.05). Conclusion: Both conventional acupuncture and conventional acupuncture plus Zhen'ai needling method can improve clinical symptoms of children with AR accompanied by AH; and conventional acupuncture plus Zhen'ai needling method has a better effect than conventional acupuncture in improving AH symptoms.

OBJECTIVETo know the anti-viral effects of rhubarb ethanol extract (REE) on herpes simplex virus(HSV) infection in vivo.

METHODSBALB/c mice inoculated from tail vein with 0.15 ml of HSV (TCID50=10(3)) were injected hypodermically with REE next day. After divided into seven groups, three groups of mice were given different doses of REE respectively and the other groups as controls. Pathological sections from the liver, spleen, kidney were made at different times of postinfection, and their pathological changes were observed under microscope; the virus titers in viscera were assayed by using plaque formation technique and the rhubarb inhibitions to the infection of HSV in vivo?were observed.

RESULTSNo toxic response to mice were observed for REE injected hypodermically; no pathological changes were observed in different therapy groups of spleens. And those in livers and kidneys at medium- and high-dosed groups disappeared quickly. The effect of low-dosed group was equal to that of positive control group, acyclovir(ACV); the results of the titer tests showed that the virus decreased rapidly by using REE, especially in the medium- and high-dosed groups which were much more marked than the low-dosed group; Q test of the data showed that total mean value had statistical significance (F=49.1459, P<0.01); moreover there were statistical significance between therapy groups (ACV, DH1, DH2, DH3) and non-therapy groups (VC) (P<0.01 ) and between DH2, DH3 and DH1 (P<0.01); no statistical significance were found between DH1, DH2 or DH3 and ACV (P>0.05). Results show that as to the effect of decreasing the average of the total titer, rhubarb is as effective as ACV; furthermore, the medium- and high-dosed groups are superior to the low-dosed group.

CONCLUSIONSREE has significant anti-viral effect on HSV in vivo; there will be a wide application foreground of it in clinical usage.

Animals ; Antiviral Agents ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Female ; Herpes Simplex ; pathology ; virology ; Herpesvirus 1, Human ; drug effects ; Male ; Mice ; Random Allocation ; Rheum

Objective To investigate the distribution of ghitathione-S-transferase M1 (GSTM1) and T1 (GSTT1) genes polymorphisms in Chinese population and smear-positive pulmonary tuberculosis cases of Jilin province. Methods Articles about GSTM1 and GSTT1 genes polymorphisms published before 2009 in China were searched. The study population was obtained from fourteen counties (or districts) of Jilin province, which included all cases from November, 2007 to May, 2008, totally 1120. The genotypes of GSTM1 and GSTT1 were detected by multiplex PCR technique. Results The frequencies of GSTM1 and GSTT1 'null' genotypes and combination M1-T1 'null' genotype acquired from systematic review were 54.2%, 46.8% and 26.2%, respectively, in Chinese Hans they were 53.4%, 44.9% and 25.5%, and in our research they are 57.2%, 20.4% and 13.7%, respectively. No significant differences between the frequencies of males and females as well as among that of different age groups were observed(P>0.05). The frequency of GSTM1 'null' genotype in our research is slightly higher than that in systematic review (P=0.016) , and the frequencies of GSTT1 'null' genotype and combination M1-T1 'null' genotype and are significantly lower than those in systematic review (both P<0.001). Conclusion The frequencies of GSTM1 and GSTTI 'null' genotypes were different among ethnics. The statistical difference between systematic review and our research may due to our large sample size and mostly Soutbern people in previous studies.

ObjectiveRecent studies revealed that the transcription factor EB (TFEB) plays an important role in regulating autophagy, reducing intracellular lipids, and inhibiting atherosclerosis. This study aims to explore the effects of atorvastatin (ATV) on autophagy and cholesterol levels of foam cells by activating TFEB.MethodsHuman mononuclear cell line THP-1 was cultured in vitro and induced to differentiate into macrophages using phorbol ester. Oxidized low-density lipoprotein (oxLDL) was added to macrophages, which were induced for 48 hours to establish a foam cell model. The experiment was divided into four groups: blank group, model group (oxLDL group), oxLDL+ Chloroquine (CQ) group, oxLDL+ ATV group, and oxLDL+CQ+ ATV group. Cells in each group were treated with drugs for 48 h. The toxicity of ATV and chloroquine on the cells was detected by the CCK8 method. Oil red O staining was used to test the level of lipid droplets. Oxidase method was used to detect levels of intracellular free cholesterol (FC), total cholesterol (TC) and others related to. Cholesterol efflux fluorescence analysis was used to determine the cholesterol efflux rate of the cells. Expression of I, P62, TFEB, LAMP 1 protein was determined by Western blot.ResultsThe results of the CCK8 method showed that the cell survival rate decreased significantly with the increase of ATV and CQ concentrations (P<0.01). Compared to the blank group, the levels of lipid droplets, FC, TC, and CE/TC in the model group significantly increased (P<0.05), and the cholesterol outflow rate significantly decreased (P<0.05). Compared to the model group, the intracellular lipid droplets, FC, TC, and CE/TC levels in the oxLDL+CQ group elevated significantly (P<0.05), while the cholesterol outflow rate decreased significantly (P<0.05). The intracellular lipid droplets, FC, TC, and CE/TC contents in the oxLDL+ATV group decreased significantly (P<0.05), and the cholesterol outflow rate increased significantly (P<0.05). Compared to the oxLDL+CQ group, the intracellular lipid droplets, FC, TC, and CE/TC content in the oxLDL+CQ+ATV group decreased significantly (P<0.05), while the cholesterol outflow rate increased significantly (P<0.05). Western blotting results showed that protein expression levels of LC3II/I, P62 and TFEB were decreased in the model histone in comparison to the blank group (P<0.05). Compared to the expression levels of LC3II/I, P62, TFEB and LAMP1 ((1.006±0.052), (0.183±0.013), (0.333±0.020), and (0.957±0.026)) in the model group, the expression levels of oxLDL+CQ histamine ((1.594±0.017), (0.257±0.006), (0.477±0.024), and (0.957±0.026)) were significantly higher (P<0.05).The protein expression levels of LC3II/I, TFEB and LAMP1 in oxLDL+ATV group ((1.146±0.060), (0.540±0.031), and (1.027±0.054)) were increased, while the protein expression levels of P62 (0.115±0.009) were decreased (P<0.05). Compared to LC3II/I and P62 in oxLDL+CQ group, the expression level of oxLDL+CQ+ATV histone was decreased ((1.419±0.036) and (0.165±0.006)), and the difference was statistically significant (P< 0.05).ConclusionATV can increase the cholesterol outflow rate of macrophages, reduce the level of cholesterol and lipid droplets in macrophages, and reduce the formation of foam cells. The mechanisms behind this are still unknown, which may be related to the activation of TFEB by ATV and influencing the process of autophagy.

Objective To investigate the clinical manifestations, MRI features and therapeutic methods of patients with central nervous system damage secondary to primary Sjogren syndrome.Methods The clinical features, MR1 manifestations and therapeutic method of 8 patients with central nervous system damage secondary to primary Sjogren syndrome, confirmed in our hospital, were retrospectively reviewed and analyzed. Results Wallenberg syndrome, limb weakness and mentality alteration appeared in these patients, some manifested as having multiple sclerosis and neuromyelitis optica, and optic nerve was frequently involved. MRI results indicated that 5 patients had lesion in the spinal cord, mainly located in the cervical and thoracic cords: 2 in the cervical cord, 2 in the thoracic cord, and 1 both in the cervical and thoracic cords; the lesions manifested as sheet lamellar, scattered plaques with mild/obvious enhancement or without enhancement; 5 patients had brain damage: lesious could be detected in the dorsolateral medulla oblongata, thalamus, hippocampus, aqueduct of midbrain,and sides of fourth ventricle and lateral ventricles, and these lesions could manifest as multiple minor ischemic focus white matter in the subcortex and cerebral ventricles, which might be similar to Wernicke encephalopathy; 1 patient appeared cervical artery dissection, and 7 patients got improvement after the treatment of hormone. Conclusion The clinical features and MRI manifestations of patients with central nervous system damage secondary to primary Sjogren syndrome are diverse.

Human Enterovirus HEV 74 is a new member of species Human enterovirus B (HEV-B). To understand its evolution and restructuring characteristics, we report the complete genome sequence of a HEV74 strain 05293/SD/CHN/2005(abbreviated as 05293) isolated from an acute flaccid paralysis (AFP) case in Shangdong Province, China, 2005. Analysis of the complete genomic sequence of 05293 showed that its genome was collinear with that of previously described 2 HEV74 strains, except for insertions and deletions at the 5'NTR and the 3 NTR regions. The complete genome sequence of strain 05293 displayed 80. 8% nucleotide and 96% amino acid identity to the prototype strain USA/CA75-10213, and 80. 6% and 95. 9% to another isolated strain Rikaze-136. The P1, P2 and P3 coding regions of strain 05293 displayed 81. 5%, 80. 0%, 79. 7% nucleotide and 95. 9%, 96. 0%, 96.2% amino acid identity to the prototype strain USA/CA75-10213, and 81. 9%, 78. 8%, 79. 5% and 95. 9%, 96. 1%, 95. 7% to strain Rikaze-136, respectively. The phylogenetic tree and Simplot analysis on 05293 and HEV-B genome sequences were performed, and the result indicated frequent recombination within HEV-B.
3' Untranslated Regions ; genetics ; 5' Untranslated Regions ; genetics ; Base Sequence ; China ; Enterovirus B, Human ; classification ; genetics ; isolation & purification ; Enterovirus Infections ; virology ; Evolution, Molecular ; Genome, Viral ; genetics ; Humans ; Muscle Hypotonia ; Paralysis ; virology ; Phylogeny ; RNA, Viral ; genetics ; Recombination, Genetic ; Sequence Alignment ; Sequence Analysis, DNA

BACKGROUNDIt is well recognized that meteorological factors have important infuences on the onset and development of many kinds of diseases. The present study was undertaken to investigate the effects of the meteorological elements on admission rates of cerebral infarction patients with hypertensive nephropathy at Changchun city, Jilin Province, northeast China.

METHODSA total of 763 medical records of inpatients from nine hospitals at Changchun city, during a period from April 6 to April 17 in 2010, were reviewed. These patients were admitted to hospitals due to the occurrence of cerebral infarction. The hypertensive nephropathy was evidenced with certain diagnosis of essential hypertension and hypertension-related kidney injuries. The cerebral infarction was diagnosed according to the World Health Organization (Stroke) standard. All the meteorological data were from practical monitoring records in Jilin Province Meteorological Observatory. The relationships between the epidemiological prevalence of cerebral infarction and meteorological variables were analyzed using the time series models of statistics.

RESULTSCompared with admission rates before the violent change in meteorological status (April 6 to April 17, 2010), the number of admission patients suffering from cerebral infarction remarkably peaked on April 12. Such an increase was highly correlated with heavy precipitation, elevation of daily average relative humidity, and reduction of average daily air temperature. With the betterment of the meteorological conditions on April 17, the admission rates of cerebral infarction patients dropped to the same level as the dates before snowing (April 6 to April 11).

CONCLUSIONSThe meteorological changes are highly associated with the occurrence of cerebral infarction in patients with hypertensive renal injury in northeast China. This study also suggested that an intensive medical interference for those patients with hypertension-induced organ injuries is very necessary in preventing the occurrence of cerebral infarction with hypertensive nephropathy when there is a violent change in meteorological condition.

Adult ; Aged ; Aged, 80 and over ; Cerebral Infarction ; China ; Female ; Hospitalization ; statistics & numerical data ; Humans ; Hypertension, Renal ; Male ; Meteorological Concepts ; Middle Aged ; Nephritis ; Patient Admission ; statistics & numerical data