AIM: To investigate the relationship between the intracellular expression of Th1 and Th2 type cytokines in placental lymphocytes and the pregnancy outcomes in NOD/SCID mice. METHODS: The resorption rate of embryos (RR) was compared between BALB/c and NOD/SCID mice. In addition, the expression rates of Th1 type (TNF-? and IL-2) and Th2 type (IL-10) cytokines were detected intracellularlly in placental lymphocytes by four-color flow cytometry. RESULTS: Although multiple-immunodeficiency was confirmed in the NOD/SCID, no significant difference was observed in RR between BALB/c and NOD/SCID mice. At the same time, a dramatically elevated level of CD8+IL-10+/CD8+ cell percentage was found at the feto-maternal interface in NOD/SCID?NOD/SCID, as compared with BALB/c?BALB/c mice, while no significant difference was observed for TNF-? and IL-2 expression in CD4+ and CD8+ cell subsets isolated from these mice. CONCLUSION: The spontaneous elevation of CD8+IL-10+/CD8+ cell percentage at the feto-maternal interface may be related to the roughly normal fertility in NOD/SCID mice.

AIM: To study the effect of ERK inhibition on the mitochondrial potential change in dexamethasone (DEX)-induced thymocyte apoptosis. METHODS: ERK activity was inhibited by PD098059 (PD), and 4 experimental groups were set: control, PD only, DEX and PD+DEX. Annexin V-FITC/PI double staining flowcytometry was used to detect apoptotic cells at time points of 3 h, 5 h and 7 h. JC-1 staining flowcytometry was adopted to examine mitochondrial membrane potential (△?m) at time points of 3 h, 7 h and 11 h. RESULTS: By stimulation with 1 ?mol/L DEX, the apoptotic rates of mouse thymocytes at 3 h, 5 h and 7 h were (19.63?0.35)%, (41.84?1.67)% and (67.00?2.43)%, respectively, and had significantly difference from control group (4.98?0.39)%, (6.08?0.33)% and (9.31?0.34)% (P0.05). At 3 h, 7 h and 11 h, the rates of low △?m cells were (21.23?1.43)%, (55.34?1.78)% and (70.88?2.87)%, significantly higher than that in control group (P0.05). CONCLUSION: DEX induces mouse thymocyte apoptosis at least partly through ERK pathway, and ERK inhibition has an important biological significance during this process.

Some patients with acute pancreatitis (AP)can relapse after initial cure. With the development and maturity of diagnosis and treatment technology,especially the rise of endoscopic technology,the detection rate of AP recurrence (such as bile duct stones,Oddi sphincter dysfunction,pancreas divisum,gene mutation,etc. )is increased. Recurrent acute pancreatitis (RAP)is characterized by various causes,and complex mechanisms. Understanding etiology and positive treatment play a pivotal role in reducing the incidence of RAP. This article reviewed the advances in study on etiology of RAP.

Objective To examine the association between uric acid (UA)levels of patients with acute ischemic stroke at ad-mission and discharged outcome.Methods The acute ischemic stroke patients of Xinganmeng People’s Hospital in Inner Mongolia,from June 1,2009 to May 31,2012 were continuity included in the present study,the included analysis sample size were 3 440 cases.Poor discharged outcome was defined as the occurrence of disability or death.With reference to the Modi-fied Rankin's Scale (MRs)Stroke Scale,Scores were recorded in the questionnaires,score of 3 or more (MRs≥3)was de-fined as disability.The patients were all grouped by P20,P60,P90 of UA,binary logistic regression were used in studying of risk factors,calculated the odds ratios (Odds ratio,OR)and 95% confidence interval (95% Confident interval,95%CI).All tests were two-sided test and a significance level of 0.05.Results A total of 359 people occurred poor outcomes in the stud-y,accounting for 10.44%.Univariate logistic regression analysis of poor outcome occurred showed that relative to the lowest group(P20,UA≤222.6 mmol/L),the second and third group (UA:222.7 ~ 310.9 mmol/L and 311.0~419.7 mmol/L) OR (95% CI)were:0.70(0.53~0.91)(P <0.05)and 0.66(0.49~0.88)(P <0.05).After adjusted age,body tempera-ture,high blood pressure,hyperglycemia,history of stroke,high triglycerides,high LDL-C and smoking,relative to the low-est level group,the second and third group occurred poor outcoming OR (95% CI)were:0.70(0.53~0.93)(P <0.05)and 0.66(0.48~0.90)(P <0.05).Conclusion Higher levels of uric acid levels in patients with acute ischemic stroke may inde-pendently related with occurred poor discharged outcome.

OBJECTIVETo evaluate therapeutic effect of Sishen Wan on experimental colitis, and explore its mechanism by expression of Bax/Bcl-2 mRNA, Fas/FasL in colonic tissue.

METHODExperimental colitis was induced by rectal administration of trinitrobenzene sulfonic acid (TNBS) dissolved in ethanol. The model animals were divided into four groups: the induced colitis but untreated group, the induced colitis groups treated with the high, middle, low dose of Sishen Wan, and the induced colitis group treated with salicylazosulfapyridine (SASP). After 10 day administration, the body weight, colonic wet weight, colonic weight index, colonic damage score and pathological change were evaluated, and the level of Fas and FasL by flow cytometry, Bax mRNA and Bcl-2 mRNA by reverse transcription polymerase chain reaction (RT-PCT).

RESULTCompared with the model group, the colonic wet weight and colonic weight index were remarkably decreased in the middle dose of Sishen Wan group (P < 0.05). The colonic injury scores were significantly reduced after rats were treated with the three doses of Sishen Wan (P < 0.05). Representative restored features were observed including fewer inflammatory cellular infiltration and follicular hyperplasia, superficial and little ulcer with fibroplasia in colonic mucosa from the treated groups. The expression of Fas in the colonic mucosa was obviously down-regulated (P < 0.05) and the ratio of Bcl-2 mRNA/Bax mRNA was significantly up-regulated (P < 0.05) in the groups treated with the three doses of Sishen Wan.

CONCLUSIONSishen Wan might postpone colonic epithelium apoptosis or improve inflammatory cell apoptosis by regulating the expression of Fas/ FasL and Bax/Bcl-2 mRNA in colonic tissue, which is possible potential path to effectively treat experimental colitis by enema.

Animals ; Colitis ; drug therapy ; metabolism ; Colon ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Fas Ligand Protein ; genetics ; Female ; Male ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; genetics ; fas Receptor ; genetics

Objective To establish the method of fingerprint and content determination of multi-component for Xiangsha Yangwei pill by gas chromatography(GC).Methods The GC fingerprint of Xiangsha Yangwei pill was found,and the peak attribution was carried out.The contents of limonene,eucalyptol,camphor,borneol,bornyl acetate,patchouli alcohol,pogostone,and α-cyperone were determined.Results The fingerprint similarity of 56 batches of Xiangsha Yangwei pill were 0.33-0.99,28 common peaks were confirmed,and 14 known components were identified.Limonene,eucalyptol,camphor,borneol,bornyl acetate,patchouli alcohol,pogostone and α-cyperone showed good linearity within the determined ranges(14.30-286.08,24.52-490.44,16.14-322.88,9.40-187.95,15.39-307.83,25.78-515.60,19.95-398.90,and 24.87-497.30 μg·mL-1).The average recoveries were 101.20％,97.90％,93.97％,94.23％,102.94％,100.54％,99.16％,and 98.31％;with the RSDs were 2.41％,1.48％,1.65％,2.00％,1.93％,2.30％,2.07％,and 2.38％,respectively.The concentrations of eight components were 0.2-959.1,0.3-420.4,1.0-542.6,0.0-64.5,0.0-364.2,0.0-339.6,0.0-130.7,0.0-82.0 μg·g-1,respectively.Conclusion The fingerprint and multi-component determination method can be used for the quality control and evaluation of Xiangsha Yangwei pill.

Objective To investigate the relationship between plasma homocysteine on admission and the outcome at discharge of acute ischemic stroke.Methods A non-concurrent cohort study was performed and a total of 1 3 1 9 patients with acute is-chemic stroke were continuously included in this study.According to tertile range of plasma homocysteine,patients were di-vided into three group.Logistic regression analysis was used to assess the independent association between plasma homocys-teine on admission and poor outcome at discharge of acute ischemic stroke.Results The difference of plasma homocysteine on admission between the poor outcome and those with good outcome had statistical significance (P<0.000 1).Without the adj ustment of multiple factors,when comparing to the first group,the second and third tertile seemed to have a tendency of increasing the risk of poor outcome at discharge,the OR (95%CI)was 2.111 (1.297~3.437,P<0.05),2.113 (1.361~3.279,P<0.05).After adjustment for multivariate,the second and third tertile also seemed to have a tendency of increasing the risk of poor outcome at discharge,the OR (95%CI)was 1.876 (1.160~3.036,P<0.05),2.396 (1.414~4.062,P<0.05).Conclusion The current study indicated that higher plasma homocysteine level was an independent risk factor for poor outcome at discharge in ischemic stroke patients.It would increase the risk of the outcome at discharge in patients with acute ischemic stroke,and suggests that there is a dose-response relationship between plasma homocysteine level on admis-sion and the poor outcome at discharge.

ObjectiveTo observe the effect of shikonin （SKN） on synovitis in DBA/1 mice with collagen-induced arthritis （CIA）. MethodThirty-six DBA/1 mice were randomly divided into a normal group， a CIA group， low-， medium-， and high-dose SKN groups （1， 2， and 4 mg·kg^-1）， and a methotrexate （MTX， 0.5 mg·kg^-1） group， with 6 mice in each group. Mice in the CIA group， the SKN groups， and the MTX group were immunized with an equal volume of bovine type Ⅱ collagen and complete Freund's adjuvant on day 1. On day 21， those mice received a second immunization with an equal volume of bovine type Ⅱ collagen and incomplete Freund's adjuvant to establish the CIA model. On the day of the second immunization， mice were treated with drugs by gavage. Mice in the MTX group received oral administration three times a week， while others received once per day， for 28 days. On day 22， the symptoms of arthritis， such as redness and swelling of joints， in CIA mice were observed， and arthritis scores were recorded. On day 49 after sample collation， histopathological examination of synovial inflammation in CIA mice was performed using hematoxylin-eosin （HE） staining. Immunofluorescence （IF） double labeling was used to detect the expression of vimentin and mitogen-activated protein kinase 1 （MAPK1） in the synovium of CIA mice. Network pharmacology predicted that the target of SKN in rheumatoid arthritis （RA） was MAPK1， which was verified by molecular docking. Western blot was used to detect the expression of extracellular signal-regulated kinase （ERK）， c-Jun N-terminal kinase （JNK）， p38， phosphorylated （p）-ERK， p-JNK， and p-p38 proteins in the synovial membrane of mice. ResultCompared with the normal group， the CIA group showed significantly higher arthritis scores， morbidity， and synovial inflammation， severely disrupted joint structure， evident articular cartilage and bone destruction， severe bone erosion （P<0.01）， increased expression of vimentin and MAPK1 in the synovium of mice， and increased protein expression of p-ERK/ERK， p-JNK/JNK， and p-p38/p38 in the synovium of mice （P<0.01）. Compared with the CIA group， the SKN groups and the MTX group showed relatively normal joint structure， with milder bone erosion and bone destruction， and smoother articular surfaces. Molecular docking results confirmed that the target of SKN was MAPK1. In the SKN groups and the MTX group， the expression of vimentin and MAPK1 in the synovial membrane was significantly reduced （P<0.01）， and the protein expression of p-ERK/ERK， p-JNK/JNK， and p-p38/p38 in the synovium of mice was significantly reduced （P<0.01）. ConclusionSKN can target MAPK1 to inhibit the protein expression of p-ERK， p-JNK， and p-p38 in CIA mice， thereby treating RA.

OBJECTIVETo analyze (CGG)n repeats sequence and AGG interspersion correlated with unstable expansion of FMR1 gene in a general Chinese population.

METHODSAmplideX FMR1 PCR Kit was used to amplify 380 X chromosomes from randomly selected 176 males and 102 females, 11 permutation carriers and 10 full mutation patients have served as controls. Results of capillary electrophoresis were analyzed with GeneMapper software Version 4.0. SPSS 11.0 software was used for statistical analysis.

RESULTSThe ratio of heterozygous females was 64.70%. The number of alleles in general males and females was 15 and 14, the classes of AGG pattern was 26 and 27, respectively. The range of alleles was between 17 to 45 CGG repeats in males and 21 to 44 CGG repeats in females, and 1 male was identified as gray zone carrier. The most frequent allele was 29 CGG repeats, which was followed by 30 and 36 repeats, while 28 CGG repeats were absent. The most common AGG pattern was 9A9A9, 99.21% of the population was detected with different forms and numbers of AGG interruption, and 6A interruption pattern was found in 10.02% samples especially in individuals with more CGG repeats. However, 57.58% of control samples had no AGG interruption, and none of the controls had 6A interruption pattern. No significant difference was observed in allele frequent distribution of (CGG)n repeats and AGG interspersion patterns between the males and females (P > 0.05), and AGGs was significantly different between general population and controls (P < 0.05).

CONCLUSIONAGGs and AGG pattern may have important roles in maintaining (CGG)n stability in general population of China, 9A9A6A9 may be a special pattern for preventing (CGG)n unstable expansion in Asian populations.

Adolescent ; Adult ; Alleles ; Female ; Fragile X Mental Retardation Protein ; genetics ; Fragile X Syndrome ; genetics ; Humans ; Male ; Middle Aged ; Trinucleotide Repeats

Objective: To investigate the efficacy and the prognostic factors in pediatric hepatoblastoma according to the standard diagnostic and therapeutic regimen. Method: Eighty-four consecutive patients were enrolled in this study between June 2000 and June 2015. Diagnosis and staging was decided by the multi-disciplinary team including oncologists, surgeons, pathologists and sub-specialized radiologists refering to protocol of Children′s Oncology Group(COG) and International Society of Pediatric Oncology Liver Tumor Study Group (SIOPEL) in a case observational study. Univariate analysis was tested by the log-rank and multivariate analysis by COX regression. All consecutive cases were divided into low risk group and high risk group according to grouping criteria. Complete remission was defined as both imaging negative and α fetoprotein (AFP) normalization. Retrospective analysis was performed in clinical features, long-term outcomes and prognostic factors. Result: Ten patients were excluded because of giving up after less than or equal to three cycles of treatment. A total of 74 cases were included in this study; 45 males and 29 females. The median age at diagnosis was 1.7 years(range 0.2-14.8 years). Untill August 30, 2016, the median follow-up time was 24.2 months (range 4.1-135.3 months); 59 cases achieved complete remission.The estimated five years overall survival (OS) and event free survival(EFS) were 90%(68/74)and 72%(58/74). AFP could be normalized after 5 circles of treatment or 2 circles of postoperation.In univariate analysis , the five years OS and EFS in low risk group were both 100%(18/18), and those in high risk group were 88%(50/56)and 68%(40/56), respectively. The five years OS rates were 75%(15/19) and 95%(53/55) in patients with or without distant metastasis (P=0.016). After 3 cycles of chemotherapy post tumor resection, we divided these patients into 2 groups according to AFP recover or not, the five years OS were 100%(43/43)and 81%(22/26), respectively (P=0.011). Conclusion The result of this protocol is reasonable when comparing with other worldwide research. Except for staging, metastasis, pathological subtypes, postoperative AFP recover or not is a prognostic factor after 3 cycles of chemotherapy.