Objective To explore the pattern of cognitive impairments mainly caused by the white matter lesions(WML) in the frontal lobe in patients with subcortical ischemic vascular disease (SIVD).Methods Fifty SIVD patients were divided into severe WML group (visual score >3, n=27) and mild WML group(visual score ≤3, n=23) according to their severities of the WML in frontal lobe .Seven patients without SIVD were collected as controls .All patients underwent a set of neuropsychological battery ,and the results were analyzed .Results There was no statistical significance among three groups on basic data .Compared with mild WML group and control group , non frontal white matter scores and numbers of lacunes in frontal lobe of severe WML group were significantly higher ( all P=0.000).Compared with mild WML group,the Montreal cognitive assessment scale in severe WML group were significantly lower ( P=0.047 ) , and scores related to the executive function were significantly lower ( P=0.006 ) , even after adjusting the numbers of lacunes in frontal lobe ,there was statistically significant difference (P=0.038). Multiple regression confirmed that the Z scores of executive functions were mainly affected by white matter lesions located in the frontal lobe ( P=0.000 ) .Conclusion WML located in the frontal lobe mainly affect the executive function in patients with SIVD .

Objective To study changes of serum IL-17 and IL-35 levels in patients with heart failure. Methods 60 patients with heart failure (observation group)were selected as research subjects.60 patients accord-ing to different severity were divided into acute period heart failure (34 cases)and stable stage heart failure (26 ca-ses);60 patients graded according to the NYHA standards were divided into 24 cases of heart failure with grade Ⅱ, 20 cases of grade Ⅲ ,16 cases of grade Ⅳ.According to the different primary diseases :expansion cardiomyopathy group (20 cases in group A),the coronary heart disease group (group B,24 cases),hypertensive heart disease group (group C,16 cases).During the same period,42 healthy elderly people in our hospital were selected as control group. The serum IL-17,IL-35 levels were tested,and the serum IL-17,IL-35 levels in patients with heart failure were ana-lyzed.Results Serum level of IL-17 in the observation group was higher than the control group,and the difference was significant [(15.61 ±4.02)pg/mL vs (9.49 ±3.96)pg/mL,t =9.018,P <0.01].Serum level of IL-35 in the observation group was significantly lower than that of the control group,and the difference was significant[(52.78 ± 4.29)pg/mL vs (61.49 ±4.81)pg/mL,t =11.963,P <0.01].The level of serum IL-17 in acute stage of patients with heart failure was higher than that of stable heart failure,and the difference was significant (t =6.278,P <0.01);IL-35 level in serum of patients with heart failure in acute phase was lower than that of stable heart failure,the difference was significant (t =9.529,P <0.01).With the increase in heart failure grade,serum IL-17 level showed a rising trend,and the differences among three groups had statistical differences (F =6.098,P <0.01);serum IL-35 level decreased,and the differences among three groups had statistical differences(F =8.978,P <0.01).The serum IL-17 level of A group was higher than that in B group and C group,there were significant differences (F =6.096, P <0.01),the serum IL-17 level between B group and C group had no statistical difference (t =0.172,P >0.05). The serum IL-35 level of A group was lower than that of B group and C group,there were significant differences (F =8.978,P <0.01),the serum IL-35 level between B group and C group had no statistical difference (t =0.208,P >0.05).Serum IL-17 and serum IL-35 level was negatively correlated (r =-0.429,P =0.009).Conclusion High expression of IL-17 in elderly patients with heart failure,while IL-35 decreased in elderly patients with heart failure, IL-17,IL-35 are closely related to the senile congestive heart failure and the severity of illness.Serum IL-17 is nega-tively correlated with the level of serum IL-35.

OBJECTIVETo study the effects of suppressor of variegation 3-9 homolog 1 (SUV39H1) gene silencing by small interfering RNA (siRNA) on the proliferation, tumor suppressor gene p15 expression and histone modification in acute myeloid leukemia cell line KG-1 cells, and to explore novel therapeutic target of leukemia.

METHODSThe SUV39H1 gene specific siRNA was synthesized in vitro and transfected into KG-1 cells by Lipofectamine(TM) 2000. The SUV39H1 mRNA and protein were detected by RT-PCR and Western blot. Cell growth affected by SUV39H1 siRNA was determined by MTS. The expressions of tumor suppressor gene p15, histone methylation of H3K9 and histone acetylation of H3, H3K9, H3K14, H3K27 and H4 were detected by Western blot.

RESULTSSUV39H1 mRNA was markedly suppressed by the SUV39H1 specific siRNA in a concentration-dependent manner. SUV39H1 siRNA inhibited the proliferation of KG-1 cells. Proliferation inhibition rate was (23.57 ± 1.98)%, (48.69 ± 1.84)%, (62.69 ± 1.61)% and (81.06 ± 3.22)% after transfected with SUV39H1 siRNA at 30, 60, 120 and 240 nmol/L for 48 hours, respectively. SUV39H1 siRNA down-regulated histone tri-methylated-H3K9 by 25%, 33% and 49% compared to control group when treated with SUV39H1 siRNA at 30, 60 and 120 nmol/L for 48 hours, while up-regulated histone acetylated H3K9 by 1.83, 2.16 and 3.07 folds, and global histone H3 in 1.35, 1.87 and 2.37 folds, but no changes were observed in histone acetylation of H3K14, H3K27 and H4. Expression of p15 increased 1.52, 2.89 and 3.08 folds after SUV39H1 siRNA treatment.

CONCLUSIONSSUV39H1 gene silencing could induce the re-expression of p15 and inhibit cell proliferation by down-regulation of histone methylation of H3K9, up-regulation of histone acetylation of H3K9 and global H3. SUV39H1 might be a new target for cancer therapy.

Cell Line, Tumor ; Gene Silencing ; Histones ; genetics ; Humans ; Leukemia ; genetics ; Methyltransferases ; genetics ; RNA, Small Interfering ; Repressor Proteins ; genetics

The total effective rate for clinical symptomstreated with oral decoction with Weiweikang(benefit-ting atrophic stomach)granules and Huoli(vitality)Bolus guided by the principle of invigorating Qi,warming the interior,activating circulation and elimi-nating blood stasis,was 91.8%,while that for thechronic atrophic gastritis with metaplasia and atypicalhyperplasia of intestinal epithelium was 87.5% and74.4% respectively.After treatment,the volume ofblood flowing was markedly increased(P

OBJECTIVETo compare the difference in the clinical efficacy on premature ovarian failure (POF) between the acupoint catgut implantation combined with artifical periodic therapy and the simple artificial periodic therapy and explore its effect mechanism.

METHODSSixty-five patients of POF were randomized into the two groups. In a western medication group, 32 cases were treated with the artificial periodic therapy with the oral administration of medroxyprogesterone acetate tablets. In a catgut implantation + western medication group, 33 cases were treated with the acupoint catgut implantation combined with artificial periodic therapy. The acupoints of Neiguan (PC 6), Zusanli (ST 36), Guanyuan (CV 4) and Sanyinjiao (SP 6) were selected. The treatment was lasted for half a year and the follow-up visit was for another half a year in the two groups. Kupperman index was used to assess the improvements in the clinical symptoms. The levels of serum sexual hormones such as follicle stimulating hormone (FSH) and estradiol (E2) were evaluated of the patients in the two groups before and after treatment. The efficacy was compared between the two groups.

RESULTSThe scores of the clinical symptoms were all significantly improved after treatment and in the follow-up in the two groups (P < 0.01, P < 0.05). In the 6-month follow-up visit after treatment, the result in the catgut implantation + western medication group was better than that in the western medication group (8.17 +/- 1.19 vs 13.68 +/- 1.08, P < 0.01). FSH was reduced after treatment in the two groups (all P < 0.01) and E2 was increased (all P < 0.05). The curative and remarkably effective rates were 75.8% (25/33) and 81.8% (27/22) after treatment and in the follow-up visit in the catgut implantation + western medication group, which were better than 67.9% (19/28) and 53.6% (15/28) in the western medication group separately (P < 0.05, P < 0.01).

CONCLUSIONThe acupoint catgut implantation combined with artificial periodic therapy achieve the remarkable improvements in the clinical symptoms of POF in the patients and the better results as compared with the simple western medication therapy. The combined therapys efficacy is stable and the long-term efficacy is apparently superior. The effect mechanism is related to the improvements in the serum sexual hormone levels.

Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Catgut ; utilization ; Female ; Follicle Stimulating Hormone ; metabolism ; Humans ; Primary Ovarian Insufficiency ; metabolism ; therapy ; Prostheses and Implants ; Treatment Outcome ; Young Adult

Objective To clone and express the encoding sequence of glyceraldehydes-3-phosphate dehydrogenase(GAPDH)from periodic Brugia molayi(Bm).Methods Total RNA was extraeted from periodic Brugic malayi.The BmGAPDH gene was amplified by RT-PCR.The PCR product was cloned and then subeloned into pcDNA3.1(+)vector.The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification,and were transformed into COS-7 cell subsequently.The expressed protein was identified by SDS-PAGE.Results BmGAPDH mRNA was highiy expressed in transfected COS-7 cell.The deduced amino acid sequence was identical with that of BmGAPDH.The recombinant pnotein wag about Nr 43 000.Conclusion The recombinant plasmid peDNA3.1(+)-BmGAPDH has been constructed and the protein has been expressed correctly.

Background Various studies have suggested that inflammatory factors such as leucocytes and macrophages are involved in the occurrence and development of diabetic retinopathy (DR),and many cytokines promote the occurrence of DR.However,the relationship of aqueous and serum monocyte chemotactic protein-1 (MCP-1) and macrophage migration inhibitory factor (MIF) change with DR is unclear.Objective This study was to investigate the effects of MCP-1 and MIF in aqueous and serum during DR development.Methods Eighty patients with type 2 diabetes were enrolled from Beijing Shijitan Hospital.These patients received phacoemulsification or phacoemulsification and vitrectomy from September,2010 to June,2011.Twenty-six cataract patients in the same stage (without diabetes) who underwent phacoemulsification surgery served as controls.According to the clinical stage of the DR,the diabetic patients were classified as the non-DR group (NDR) (20 eyes),non-proliferative DR group (NPDR) (38 eyes) and proliferative DR group (PDR) (22 eyes).Aqueous humour and periphery blood samples were collected during the operation to detect MCP-1 and MIF using enzyme-linked immnunosorbent assay (ELISA).Written informed consent was obtained from each subject before any relevant medical examination.Results The average aqueous MCP-1 levels were(1660.78±562.98),(1463.26± 623.41),(686.76±186.16) and(494.35±148.59) ng/L in the PDR group,NPDR group,NDR group and control group,respectively,showing a significant difference among the 4 groups (F=37.968,P=0.000).No significant differences were found in the aqueous MCP-1 levels between the control group and NDR group (P=0.169),or between the NPDR group and PDR group (P=0.117).However,the aqueous MCP-1 levels were significantly elevated in the PDR group,NPDR group and NDR group compared with the control group (P=0.000).The average aqueous MIF levels were (6.85±1.99),(3.56±0.90),(1.10±0.48) and (0.86 ± 0.46) μg/L,respectively,with significant differences among them (F =144.502,P =0.000).Multiple comparisons between groups were found to be significantly different (P =0.000) according to the LSD-t test,except between the control group and NDR group (P =0.475).A significant positive correlation was seen between the aqueous MCP-1 level and MCP-1 level in all study participants (r =0.564,P =0.000).However,serum levels of MCP-1 and MIF were not statistically significantly different among the 4 groups (F =2.158,P＞0.05;F =0.813,P＞0.05).Conclusions The increase of the aqueous MIF and MCP-1 levels is associated with the progression of diabetic retinopathy.The results suggest that MIF and MCP-1 promote the occurrence of DR.

OBJECTIVETo investigate the immunological effect of PM2.5 on cytokine production in female Wistar rats.

METHODSFemale Wistar rats were given 0.3 mg, 0.75 mg, 2 mg, 5 mg of PM2.5 per 0.5 mL saline, respectively. Saline was used as the negative control. TNF-alpha and IL-6 levels in the branchoalveolar lavage were measured by ELISA, and mRNA expression levels in lung tissue were detected by RT-PCR. Alveolar macrophages were collected for testing phogacytic function.

RESULTSExposure to PM2.5 stimulated TNF-alpha production in a dose-dependent manner (P < 0.05), However, no statistically significant difference was found. No time-dependent change in TNF-a and IL-6 production was found. TNF-alpha and IL-6 mRNA expressions were induced by PM2.5-exposure. The phagocytic rate (PR) was significantly decreased by PM2.5 treatment.

CONCLUSIONPM2.5 exposure increases inflammation response of the lung in a dose-dependent manner. Moreover, tissue injury induced by PM2.5 may be related to altered production of cytokines. PMz2.5 may impair the phagocytic activity of alveolar macrophages.

Animals ; Bronchoalveolar Lavage Fluid ; Cytokines ; biosynthesis ; genetics ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Female ; Macrophages, Alveolar ; metabolism ; Particle Size ; Phagocytosis ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo explore the cytotoxic responses of spleen T lymphocytes (CTL) in BALB/c mice induced by recombinant HSP110-HER2/neu ICD complex.

METHODSTumor-bearing mouse model was immunized by HSP110-HER2/neu ICD complex. The IFN-γ level secreted by activated spleen T lymphocytes was detected by enzyme linked immunospot assay (ELISPOT). The corresponding CTL activity was measured by granzyme release assay.

RESULTSThe BALB/c mouse model of human mammary tumor highly expressing HER2/neu was established. HSP110-HER2/neu ICD complex immunization led to a significantly higher level of INF-γ than that in HSP110-P(789-797) immunized and HER2/neu ICD immunized mice. HSP110-HER2/neu ICD complex immunized animals also show significant CTL activity. The results of immunohistochemical staining showed that the number of blue spots in the PBS group was 4.57 ± 1.33, HSP110 group 6.83 ± 2.08, HER2/neu ICD group 16.17 ± 2.86, HSP110-P(789-797) group 43.67 ± 4.78, and SP110-HER2/neu ICD group 76.51 ± 8.17. The number of IFN-γ-secreting spleen lymphocytes in the HSP110-HER2/neu ICD group was significantly higher than that in the HSP110-P(789-797) group, and that of HSP110-P(789-797) group was significantly higher than that of HER2/neu ICD group (P < 0.01). The target cell-killing rate of the PBS group was (8.15 ± 1.27)%, HSP110 group (9.51 ± 1.51)%, HER2/neu ICD group (14.03 ± 2.45)%, HSP110-P(789-797) group (25.99 ± 3.04)% and HSP110-HER2/neu ICD group (38.15 ± 3.95)% (all P < 0.01).

CONCLUSIONSHSP110-HER2/neu ICD complex can promote the proliferation and maturation of T lymphocytes into CTLs, and might be used as anti-tumor vaccine to induce potent cytotoxic T lymophocyte immunoresponse against specific tumor cells.

Animals ; Breast Neoplasms ; metabolism ; pathology ; Cancer Vaccines ; immunology ; Cell Line, Tumor ; Cell Proliferation ; Female ; HSP110 Heat-Shock Proteins ; immunology ; Humans ; Interferon-gamma ; metabolism ; Lymphocyte Activation ; Mice ; Mice, Inbred BALB C ; Neoplasm Transplantation ; Random Allocation ; Receptor, ErbB-2 ; immunology ; metabolism ; Recombinant Proteins ; immunology ; Spleen ; cytology ; immunology ; T-Lymphocytes ; cytology ; immunology ; metabolism ; T-Lymphocytes, Cytotoxic ; immunology ; Vaccines, Synthetic ; immunology

BackgroundMonocyte chemotactic protein-1 (MCP-1)plays an important role in the tumor,inflammation,diabetic retinopathy and other neovascular disease,but the expression and the role of MCP-1 in the oxygen induced retinopathy(OIR) model have rarely been reported. Objective This study was to investigate the expression of MCP-1 in the retina development of newborn mouse and in mouse models with OIR.Methods C57BL/6J newborn mice were divided into two groups and 60 mice in each group.Mice in OIR group were exposed to 75％ oxygen for 5 days and then to room air.All mice in normal control group exposed to room air only.Ten mice in each group were randomly chosen and sacrificed at postnatal 5,7,12,14,17,21 days.The expression of MCP-1 in mouse retina was detected with the method of immunohistoehemistry and reverse transcription polymerase chain reaction(RT-PCR).Results MCP-1 positive cells were seen in normal mouse retina.Up-regulation of MCP-1 positive cells was detected both in 12 days in normal control group and in 14 days in OIR group.MCP-1 mRNA was detected in mouse retina at 5 days,and a transient up-regulation of MCP-1 mRNA was observed in 12 days in normal control group.MCP-1 mRNA in OIR group significantly increased in 14 days in comparison with the normal control group( P =0.028,P =0.001 ). Conclusions Expression of MCP-1 is detectable in whole retinal development procession of mice.A transient up-regulation of MCP-1 expression is detected in the critical period of retinal vascular development in mice models with OIR,which is closely related to the retinal vascular development and progression of retinal new vessels.