Objective To investigate the effects of recombinant human tumor necrosis factor receptorⅡ-Fc fusion protein(rhTNFR:Fc ) on nuclear factor-κB(NF-κB)and tumor necrosis factor-α(TNF-α)protein expression in the rat brain tissue,and traumatic brain edema following acute traumatic brain injury(TBl).Methods The inju-ry rats were subjected to right lateral cortical impact injury caused by a free-falling object.In the rhTNFR:Fc group,rhTNFR:Fc was administered intraperitoneally(3.0 mg/kg)after 30 min of injury-and the rats of control and injury group administered with normal saline solution.The levels of TNF-α protein in rat brain were mensured by radio-im-munonssay(RIA).The NF-κB protein expression of rat brain was studied immunohistochemically.In the meantime,the water content of rat brain Was measured,and for the microscope and ultrastructure examinations by using electron-ic microscope respectively.Results Compared with control group,the levels of NF-κB and TNF-α protein and the water content of brain tissue were obvicously increased from 6 h following TBI(P<0.05,or P<0.01).Compared with injury group,the expression of NF-KB and TNF-α protein,and the brain water content were lower in rhTNFR:Fc group(P<0.05-P<0.01).The rhTNFR:Fc Can reduce the brain injury in electronic microscope examinations.Conclusion The expression of NF-κB and TNF-α,and the water content of rat brain increase significantly follow-ing acute traumatic brain injury.The rhTNFR:Fc can dramatically inhibit the expression of NF-κB and TNF-α,and alleviate rat brain edema after TBI.

Objective To investigate the effect and mechanism of extract ginkgo biloba (EGb) on learning memory ability and hippocampus CA1 region Caspase-9P35 activity in dementia rats induced by D-gal.Methods 48 rats were randomly divided into six groups.The dementia model were established by injecting intraperitoneally with the D-gal and each EGb group was injected different doses of EGb simultaneously.TUNEL method was used to detect apoptosis of hippocampal neurons in the CA1 region.Immunohistochemistry and Western Blot were used to determine the expression of Caspase-9P35 in hippocampus CA1 region.Results Compared with the normal group,TUN EL-positive neurons ( (37.8 ± 1.3 ),(0.8 ± 0.2 ) ) and the activity of Caspase-9P35 ( (37.6 ±1.8 ),(6.2 ± 1.3 ) ) had significant increase in hippocampal CA1 subfield of D-gal group (P ＜ 0.01 ).Contrast to D-Gal group,TUNEL-positive neurons ( ( 17.6 ± 0.9),(9.8 ± 0.8 ),( 37.8 ± 1.3 ) ) and the activity of Caspase9P35( (28.6 ± 1.3),(25.0 ± 1.6),(37.6 ± 1.8) ) were significant decreased in EGb-M and EGb-H group (P＜0.01 ).While TUNEL-positive neurons and the activity of Caspase-9P35 had not significant difference in the therapy group than D-Gal group (P ＞ 0.05).Conclusion EGb can improve the cognitive level of the dementia rats.One of the therapeutic mechanisms may be to decrease the hippocampus CA1 region Caspase-9P35 activity.The results of the pretreatment group was more effective than the therapy group.

Recent data have revealed that inhibiting autophagy exacerbates lipid accumulation in hepatocytes and nitrite treatment reduces total triglyceride levels in the high-fat diet mice. Therefore, the present study aimed to determine the effects of nitrite on simple hepatic steatosis and the possible role of autophagy. Firstly, steatotic L-02 cells were induced by incubating L-02 cells with 1.2 mmol · L(-1) oleic acid (OA) for 24 h. Secondly, steatotic L-02 cells were treated with 0.2 mmol · L(-1) sodium nitrite (SN) plus 3-methyladenine (3-MA), or chloroquine (CQ) for 24 h, and then lipid accumulation was measured with oil red O staining and triglyceride quantification. The notable steatosis could be observed in L-02 cells following exposure to 1.2 mmol · L(-1) OA for 24 h. Treatment with 0.2 mmol · L(-1) sodium nitrite reduced lipid accumulation in steatotic L-02 cells. 3-MA weakened the ability of sodium nitrite to ameliorate hepatic steatosis. Additionally, the sodium nitrite increased number of LC3-II immunostaining puncta and LC3-II protein expression was confirmed by immunofluorescence or Western blot analysis, and the effects were enhanced by CQ treatment. The number of increased cytoplasm vacuoles and lysosomes increased was confirmed by phase contrast and fluorescence microscope respectively. The increased autolysosome was detected by electron microscopy, this phenomenon could be reversed by CQ treatment. These data demonstrated that sodium nitrite enhanced the autophagic flux and decomposition of triglycerides in steatotic L-02 cells.

Objective To investigate the impact of AG490 on the blood-brain barrier (BBB ) permeability and the expression of interleukin-6 (IL-6 )and tumor necrosis factor-α(TNF-α)after traumatic brain injury (TBI)in rats. Methods A total of 144 healthy male SD rats were randomly divided into a control group,a trauma group,and an AG490 intervention group (n=48 in each group). The rats in each group were redivided into four subgroups (4 h,1 d,3 d,and 7 d subgroups)according to the time points after cerebral injury (n=12 in each subgroup). A brain trauma models were induced by hydraulic shock method. Evans blue was used to determine the changes of the BBB permeability after cerebral injury in each group. Real-time fluorescence quantitative PCR was to detect the expression levels of TNF-αand IL-6 mRNA in rat brain tissue. Immunohistochemistry was used to detect the expression of human phospho tyrosine kinase (P-JAK2). Results (1)The permeability of BBB:The permeability of BBB increased at 4 h,1 d,3 d and 7 d after brain injury in the trauma group (Evans blue permeation:10. 4 ± 1. 2,16. 0 ± 1. 4,22. 3 ± 2. 0,and 8. 4 ± 0. 9μg/g,respectively). Compared with the control group, there were significant differences (all P<0. 01). The Evans blue permeation of the AG490 intervention group were 9. 1 ± 1. 0,12. 8 ± 1. 1,17. 5 ± 1. 4 and 7. 1 ± 0. 8μg/g,respectively at each time point,and they were all significantly lower than those of the trauma group (all P<0. 01). (2)The expression of IL-6 and TNF-α mRNA:The expression levels of IL-6 mRNA and TNF-α mRNA at 4 h,1 d,3 d and 7 d after traumatic brain injury in the trauma group were 2. 31 ± 0. 35,2. 73 ± 0. 35,3. 32 ± 0. 29,2. 14 ± 0. 24 and 7. 46 ± 1. 18,9. 42 ± 1. 54,13. 76 ± 1. 89,and 6. 28 ± 1. 00,respectively,they were all significantly higher than those of the control group (all P<0. 01). The expression levels of IL-6 mRNA and TNF-α mRNA of the AG490 intervention group were 1. 14 ± 0. 22,1. 54 ± 0. 23,1. 94 ± 0. 32,1. 26 ± 0. 21 and 5. 57 ± 0. 88, 7. 78 ± 1. 02,11. 51 ± 1. 29,and 5. 05 ± 0. 97,respectively,they were all lower than those of the trauma group,but they still higher than the control group. There were significant differences (all P<0. 01). (3 )The expression of P-JAK2:The expression levels of P-JAK2-positive cells at each time point after traumatic brain injury in the trauma group were significantly higher than the control group (all P<0. 01),they were 17. 4 ± 2. 7,56. 2 ± 6. 7,26. 1 ± 5. 4,and 15. 3 ± 2. 5,respectively;those of the AG490 intervention group were 12. 2 ± 1. 4,41. 5 ± 4. 6,19. 4 ± 4. 1,and 9. 6 ± 2. 0,respectively,they were all lower than those of the trauma group,but still higher than the control group. There were significant differences (all P<0. 01). Conclusion During the acute phase after TBI,AG490 may activate the factor signaling pathways by inhibiting the non-receptor tyrosine kinase/signal transduction and transcription,significantly inhibit the expression of brain tissue inflammatory cytokines IL-6 IL-6 and TNF-α,reduce the BBB damage,and help to reduce secondary brain injury.

A precise and selective reversed phase high-performance liquid chromatographic method (HPLC) was used to quantify the levels of huperzine A in samples of three Huperzia serrata populations with a total of 73 individuals located in Zhejiang, Guangxi, Chongqing, respectively, as well as in one-to-one samples of these 73 individuals introduced in same site after one year. Huperzine A content variation both among and within populations, and the dynamic change of this alkaloid occurring in same population after one year introduction, were analyzed using SPSS 13.0 software (Coefficient of variation, One-way ANOVA analysis, Paired-samples T tests). The results indicated that huperzine A content varied significantly by geographical locations, especially change with longitude, i. e., the order of the huperzine A content was CQ population > GX population > ZJ population. The coefficients of variation (CV) were as follows: 0.36 (CQ), 0.44 (GX) and 0.40 (ZJ). This indicated that there was plentiful diversity concerned with huperzine A content among individuals within population. Moreover, this high diversity was still maintained after one year introduction. ANOVA analysis showed that there was significant difference among populations in huperzine A content. Finally, the significant change of huperzine A content was not observed in all three populations after one year introduction. The results presented in this study could provide evidence that the huperzine A content variation of H. serrata is the results of an interaction between genes and the environment, by comparison, is mainly controlled by genetic factor.
Chromatography, High Pressure Liquid ; methods ; Chromatography, Reverse-Phase ; methods ; Drugs, Chinese Herbal ; Huperzia ; chemistry

Aim To investigate the effect of hydrogen sulfide on hepatic lipid accumulation in obese mice. Methods C57 BL/6 J mice were randomly divided into control group, model group, and NaHS group. The mice of the control group were fed with normal diet. The mice of the model group and the NaHS group were fed with high-fat diet. From the thirteenth week, the mice of NaHS group were injected intraperitoneally with NaHS (H2S donor) in a dose of 50 μmol·kg-1 per day for 4 weeks and the mice of the model group were injected with the same volume of saline. All mice were sacrificed at the end of the 16th week. The tis-sues of liver were homogenized and centrifugated. The supernatants were used for the determination of triglyc-eride and cholesterol in liver. The morphology of liver was tested by H&E staining. Liver lipid accumulation was determined by oil red staining. Total RNA was ex-tracted from frozen tissue of liver. PCR was used to de-tect CPT-1 , FAS gene expression and ELISA method was used to detect CPT-1,FAS activity in mice liver. Results The body weight of the mice from NaHS group and model group was bigger than that of the mice from control group. Compared with the model group, the body weight of the mice from NaHS group was less;the content of triglyceride and cholesterol in liver was lower; the degree of liver tissue pathological changes and lipid accumulation were alleviated; CPT-1 expres-sion and activity were increased; FAS expression and activity were decreased. Conclusions These data in-dicate that hydrogen sulfide can reduce the lipid con-tent of liver tissue in obese mice and alleviate fatty liv-er. The mechanism may be associated with the in-creased expression of CPT-1 and the decreased expres-sion of FAS in liver.

Objective To analyze the discrepancy and similarities of clinical characteristics in elderly patients with ulcerative colitis (UC) and ischemic colitis (IC).Methods A total of 43 elderly patients (age≥60 yrs) with UC and 36 elderly patients with IC were enrolled from 2004 to 2015 at Peking Union Medical College Hospital.The clinical characteristics were retrospectively analyzed and compared between the two groups.Results Compared with IC group,the disease course was longer with lower incidence of cardiovascular comorbidities in UC patients (P ＜ 0.05).In UC group,more patients presented with diarrhea,mucopurulent bloody stool [39 (90.7％) vs 16 (44.4％) and 34 (79.1％) vs 2 (5.6％)respectively,both P ＜ 0.01].Yet bloody stool as the only symptom was seen in more IC patients than UC patients [61.1％ (22/36) vs 7.0％ (3/43),P ＜ 0.01].The ratio of extra-intestinal manifestations was higher [18.6％ (8/43) vs 0 (0/36)] in UC patients,while complications were lower [11.6％ (5/43) vs 30.6％ (11/36),P ＜ 0.05].As to the laboratory parameters,median platelet count [(294.38 ± 104.83) × 109/L vs (235.47±94.82) × 109/L,P ＜0.05] was higher in UC group.In addition,more patients with UC had positive perinuclear antineutrophil cytoplasmic antibody (p-ANCA) [50.0％ (15/30) vs 10.0％ (2/20),P ＜ 0.05].The most commonly involved regions of IC were descending colon and sigmoid colon,in which the lesions were clearly demarcated with the normal mucosa.Lesions in patients with UC mainly originated from rectum and might spread to the whole colon.Vascular occlusion and micro thrombosis were characteristic pathological findings of IC.The crypt abscesses were frequently seen in the UC group.Conclusion Even though UC and IC have some similar manifestations in the elderly patients,clinical and pathological discrepancy is still helpful to differentiate each other.

OBJECTIVETo study the difference in stress and apoptosis related genes transcription between hTERT-RPE1 cells exposed to simulated microwave radiation and the cells with heat water bath, and the effects of microwave on gene transcription in cultured human retina pigment epithelial cells.

METHODScDNA microarray technique was used to detect the mRNA isolated from hTERT-RPE1 cells exposed to 2 450 MHz simulated microwave radiation and with heat water bath, respectively.

RESULTSAmong the 97 related aim genes, there were seven genes up-regulating its transcription, i.e., M31166 (2.52fold), L24123 (2.66fold), AF039704 (2.22fold), U67156 (2.07fold), AF040958 (2.13fold), NM-001423 (2.63fold) and NM-005346 (3.68fold). But, no notably down-regulating gene in transcription was detected.

CONCLUSIONSMicrowave could induce up-regulating in multiple stress and apoptosis related genes transcription in cultured human retina pigment epithelial cells, hTERT-RPE1 cells. Microwave radiation has unique effect itself in addition to its heat effect.

Gene Expression Profiling ; Gene Expression Regulation ; radiation effects ; Humans ; Microwaves ; Oligonucleotide Array Sequence Analysis ; methods ; Pigment Epithelium of Eye ; cytology ; metabolism ; radiation effects ; Retina ; cytology ; metabolism ; radiation effects ; Time Factors

Objective To analyze the clinical manifestations, imaging features, histopathology and genes of cerebrotendinous xanthomatosis (CTX) to improve the understanding of clinical workers on the disease. Methods The imaging examination, histopathological and gene detection methods were synthetically applied, and the disease characteristics of two patients with CTX, admitted to our hospital in March 2018 , were analyzed. Results (1) Patient one was a 52-year-old male, with typical neurological symptoms: weakness of both lower limbs, damage of pyramidal tract and extrapyramidal system, cognitive impairment, and ataxia; non-neurological symptoms included cataract, arch foot, and Achilles tendon mass; cranial MR imaging indicated symmetrical abnormal signal of cerebellar dentate nucleus, low signal on T1WI, and slightly high and low signal intensity on T2WI/FLAIR; achilles tendon biopsy showed fibrous connective tissues with multiple xanthoma cells and multinucleated giant cells aggregation, accompanied by cholesterol crystallization. (2) Another 16-year-old male presented with cerebellar ataxia, recurrent seizures, mental and motor retardation, and congenital cataract; two heterozygous mutations of CYP27A1 gene were detected by gene detection; and the mutations were c.373-379 delCCAGTAC and c.1420C>T. Conclusion The clinical manifestations of CTX are varied; early imaging examination lacks specificity; it can be clearly diagnosed by histopathology and CYP27A1 gene detection.

Objective:To investigate the long-term effect of the implantation of human umbilical cord-derived mesenchymal stem cells (HUC-MSCs) for type 1 diabetes mellitus.Methods:Fifteen patients with type 1 diabetes mellitus were treated with HUC-MSCs from September 2009 to December 2011 at Department of Endocrinology and Metabolism, the Second People′s Hospital. Patients were followed-up for 10 years and the parameters were collected including fasting blood glucose, HbA 1C, mean amplitude of glycemic excursions (MAGE), fasting C-peptide, daily insulin doses and glutamic acid decarboxylase antibody (GADA). Results:Among 15 patients, 1 patient (6.67%) was found with breast cancer. All patients with type 1 diabetes mellitus decreased daily insulin doses due to frequent hypoglycemia one week later. Six months later, 4 patients (26.67%) stopped insulin injection. While among the 4 patients, 1 patient (6.67%) had not yet used insulin until today and GADA was negative, the other 3 patients (20.00%) restarted insulin within 3-5 years after implantation with significantly less daily insulin doses [(18.00±1.00)U vs (29.00±1.73)U, P<0.01]. The remaining 11 patients (73.33%) with type 1 diabetes mellitus who did not stop insulin also had significantly lower daily insulin doses [(18.09±0.83)U vs (29.64±0.89)U, P<0.01]. The level of MAGE was signicantly decreased compared to those of pre-implantation [(6.14±0.25)mmol/L vs (9.72±0.32)mmol/L, P <0.01], while fasting C-peptide level was significantly improved[(0.91±0.03)nmol/L vs (0.11±0.01)nmol/L, P <0.01]. There were no significant differences in fasting blood glucose and HbA 1C before and after implantation. Conclusions:The implantation of HUC-MSCs for the treatment of type 1 diabetes mellitus can restore the function of islet β cells, decrease daily insulin doses and reduce blood glucose fluctuations in the long term. Although precise mechanisms are unknown, this therapy is expected to be an effective strategy for treatment of type 1 diabetes mellitus.