Objective: To explore the extraction process of Trollii Flos Oral Liquid (TFOL). Methods: HPLC was used to determine orientin and vitexin in liquid extract of TFOL, using mass concentration of both as index. The orthogonal design L18(37) and reverse continuous cycle cryogenic extraction were used to extract Trollii Flos, the optimum extraction was determined, compared with the current standard of TFOL extraction process. Results: The optimum extraction conditions of reverse continuous cycle cryogenic extraction were as follows: extraction temperature 90°C, extraction time 2 h, powder type 10 mesh, added of 20 times water. The linear ranges of orientin and vitexin were 18.46-295.37 (r = 0.9999) and 5.84-93.44 μg/mL (r = 0.9999), the average recoveries of orientin and vitexin were 96.07%, RSD = 0.74% (n = 6), and 97.73%, RSD = 1.42% (n = 6), respectively. Conclusion: The technology of reverse continuous cycle cryogenic extraction is stable and feasible, and the extraction rate is high; The measurement method is accurate, sensitive, simple, and stability, and has good reproducibility. The extraction and determination methods could be used as production process and quality control reference of TFOL.

Objective To explore the survival mechanism of hippocampal ne urons after damage of hypoxia-ischemia and reperfusion of brain.Methods Seven days old SD rats(n=56) were randomly divided into hypoxia-ischemia br a in iniury(HIBD) group and sham group.The HIBD and reperfusion model was establis hed.The flowing of blood was de tected by multicolor Doppler.The p-CREB(phosphorylated c-AMP response element bi nding protein)and c-Jun were immunohistochemically evaluated in hippocampus.Thi onin staining was used to observe the apoptosis.Results The expression of p-CREB reache d the peak at 3,24 h postreperfusion in the right hippocampus of HIBD group,and then decreased to the normal level on the 7th day.In contral group the same reg ions showed basic immn-noreactivity.While c-Jun reached the peak at 6 h postreperfusion,then with a slightly decrease at 24 h;and at 48 h the other peak appeared,then with a gradual decline .On the 7th day the mumber of positive cells were still significanthy more than control group(P0.05).The sham animal showed very few apoptosis cells in the regio ns of hippocampus.Conclusions The persistent activation of CREB in the hippocampus regulates,the expression of c-Jun through the signal transductions and is involved in the course of neuron s′ survival and repair during the period of post hypoxia-ischemia reperfusion.I t is very important for the protection of the pyramidal hippocampal neurons on t he damaged side,especially for the sensitive region CA1. J Appl Clin Pediatr,2005,20(2):133-135

OBJECTIVETo identify the expression characteristics of the 1700001022RIK (RIKEN cDNA 1700001022) gene in mice and explore its function by bioinformatic analysis.

METHODSUsing the expression profile of gene microarray, we detected the expression of a new testis-specific gene, 1700001022RIK, in mice. We analyzed its expression characteristics in the testis tissue and their changes in different developmental stages of the testis by RT-PCR, real-time RT-PCR, Western blot, and immunohistochemistry. We performed bioinformatic analysis using a bioinformatic software.

RESULTSThe 1700001022RIK gene was specifically expressed in the mouse testis in an age-dependent manner, most highly in the adult mice. The 1700001022RIK protein was mainly expressed in the spermatogonia, spermatocytes, and round spermatids of the adult mice. Bioinformatic analysis showed that the 1700001022RIK protein amino acid sequence had a high similarity in human and mice, which indicated that this gene was highly conserved in mammals.

CONCLUSION1700001022RIK is a testis-specific gene mainly expressed in the spermatogonia, spermatocytes, and round spermatids of seminiferous tubules, which might be involved in the regulation of spermatogenesis.

Age Factors ; Animals ; Blotting, Western ; Computational Biology ; DNA, Complementary ; Gene Expression ; Genomics ; Male ; Mice ; Molecular Chaperones ; genetics ; Seminiferous Tubules ; Spermatids ; Spermatocytes ; Spermatogenesis ; genetics ; Spermatogonia ; Testis

Animals ; Animals, Newborn ; Cyclic AMP Response Element-Binding Protein ; analysis ; Hippocampus ; blood supply ; chemistry ; pathology ; Hypoxia-Ischemia, Brain ; physiopathology ; Immunohistochemistry ; Proto-Oncogene Proteins c-fos ; analysis ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; physiopathology ; Time Factors

Objective To find out the iodine nutritional status and thyroid function of pregnant and lactating women in the urban areas of Wuwei City of Gansu Province,and to provide a scientific basis for iodine supplementation.Methods Blood and urine samples of pregnant (52 persons) and lactating women (59 persons) were collected in 2009.Urinary iodine content was measured with arsenic cerium catalytic spectrophotometric.Serum free three triiodothyronine(FT3),free thyroid hormone(FT4),triiodo thyronine(TT3),serum total thyroxine (TT4) and thyroid-stimulating hormone (TSH) were detected with direct chemiluminescence immunoassay.Results The medians urinary iodine of pregnant and lactating women were 274.68,217.88 μg/L.The rates of low urinary iodine (pregnant women below 150 μg/L,lactating women lower than 100 μg/L) were 9.62％ (5/52) and 6.78％ (4/59).Serum TT3,TT4 levels in pregnant women[(2.48 ± 0.59),(132.18 ± 33.36)nmoL/L] were higher than that in the lactating women[(2.16 ± 0.49),(108.79 ± 28.36)nmol/L,t =-3.123,-3.971,all P ＜ 0.05].Thyroid dysfunction incidence rates of pregnant and lactating women were 17.31％ (9/52) and 8.47％ (5/59).Thyroid dysfunction was mostly subclinical hypothyroidism.Conclusions Overall iodine nutrition of pregnant and lactating women is in good condition,some individuals have the trend of hypothyroidism.It is necessary to carry out routine monitoring of urinary iodine and thyroid function.

Objective： To investigate the effect of oxLDL and VitE on the expression of CD40 and CD40 ligand(CD40L) in cultured human monoc ytes. Methods： The expression of CD40 and CD40L on monocytes su rface were measured by indirect immunorescence technique in combination with flo w cytometry. Results： Low concentration of oxLDL（≤200 μg/L） significantly increased the expression of CD40 and CD40L in a dose and time dep endent manner. High concentration (>200 μg/L）of oxLDL markedly reduced the exp ression of CD40 and CD40L. When VitE was added, it significantly reduced the ex pression of CD40 and CD40L on monocytes surface induced by oxLDL in a dose-depe ndent manner. Conclusion：It is an important mechanism that the high expression of CD40 and CD40L induced by oxLDL may be contributed to the for mation of atherosclerosis. Antioxidan VitE can partially inhibit the high expres sion of CD40 and CD40L on monocytes surface induced by oxLDL.

Tumor immune checkpoint therapy is a clinical treatment strategy developed based on the new principle of the inhibition of negative immune regulation. In this article, the tumor immune checkpoint therapy and the drug delivery strategies were reviewed, mainly including immunity and tumor therapy, tumor immune checkpoint therapy and its mechanism of action, clinical application of tumor immune checkpoint therapy and therapeutic drugs, immune resistance of programmed cell death protein 1 (PD1)/programmed cell death ligand 1 (PDL1) treatment and countermeasures, drug delivery strategies for tumor immune checkpoint therapeutic agents, etc. As a revolutionary new immunotherapy strategy, tumor immune checkpoint therapy has shown obvious superior therapeutic efficacy in a variety types of tumor. However, tumor immune checkpoint therapy is also faced with a big challenge, namely, immunotherapy resistance. With the discovery of new mechanism, the continuous development of new therapeutic drugs and delivery strategies, tumor immune checkpoint therapy is expected to further improve the clinical efficacy of tumor.

OBJECTIVETo investigate the effect of adenosine A2A receptor knockout (A(2A)RKO) on relationship between continuous activation of phospho-c-Jun N-terminal kinase (P-JNK) and expression of nerve cell apoptosis in hippocampus CA1 domain of newborn mice after hypoxia/ischemia brain damage(HIBD) and its potential mechanism.

METHODSA(2A)RKO mice and adenosine A2A receptor wildtype (A(2A)RWT) littermates (n = 80) were divided into Sham operation group (S) and model group (M), 1, 3 and 7 day after HIBD, totally 8 groups. HIBD was developed with 7 day-old neonatal mice according classical Rice-Vannucci method. It was tested the effect of A(2A)RKO on short-term neurofunctional outcomes consisted of three developmental reflexes (righting, geotaxis and cliff aversion), the changes of brain pathology with hematoxylin-eosin (HE) staining and Nissl staining, the expressions of nerve cell apoptosis with terminal deoxynucleotidyl transferase mediated dUTP-biotin nick-end labeling(TUNEL) staining and P-JNK were observed by immunohistochemistry.

RESULTSThe neurological behavior injuries and brain histopathological damages and nerve apoptosis cells were aggravated in A(2A)RKO newborn mice after HIBD. The positive expressions of P-JNK were significantly higher in the ischemic hippocampus CA1 domain after HIBD than ones in group S respectively (P < 0.01), reaching to peak at 1 day and then began gradually decreasing. P-JNK expression in model knockout(MKO) at 1, 3 and 7 day increased greatly compared to those in the previous time point of corresponding model wildtype (MWT) (P < 0.01, P < 0.05, P > 0.05); there was a positive correlation between the expressions of P-JNK and nerve cell apoptosis after HIBD in newborn mice(r = 0.837, P < 0.01).

CONCLUSIONEarly continuous activation of P-JNK might be involved in the aggravated nerve apoptosis cells and brain damage induced by A(2A) RKO newborn mice after HIBD.

Animals ; Animals, Newborn ; Apoptosis ; Hypoxia-Ischemia, Brain ; metabolism ; pathology ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Mice ; Mice, Knockout ; Neurons ; drug effects ; metabolism ; pathology ; Receptor, Adenosine A2A ; genetics

Objective： To investigate the effect of c ytokines (IFN-γ，TNF and IL-1) on the expression of CD40 and CD40 ligand (CD4 0L) in monocytes/macrophages. Methods： The mRNA expression of C D40 and CD40L was measured by RT-PCR and the CD40，CD40L expression on the mono cytes/macrophages were detected by flow cytometric analysis. Results： IFN-γ，TNF and IL-1 could not only significantly up-regulate the mRNA levels of CD40 and CD40L in cultured monocytes/macrophages, but also increase t he expression of CD40 and CD40L. Antioxidant VitE could reduce the expression o f CD40 and CD40L induced by IFN-γ，TNF and IL-1. Conclusion： IFN-γ，TNF and IL-1 can stimulate high expression of CD40 and CD40L . Antio xidant VitE can partially inhibit the expression of CD40 and CD40L induced by cy tokines in cultured monocytes/macrophages.

Objective: To investigate the effects of hyaluroni dase (HAase) and hyaluronan (HA) on proliferation of vascular endothelial cells and its mechanism. Methods: The cultured bovine aortic endothel ial cells (BAEC) were treated with HAase or HA. Cell proliferation rate was dete cted by MTT assay. The expression of CD44 and DNA content of the cells were meas ured by flow cytometry (FCM). Results: HAase (50 μg/ml) stimula ted cell proliferation ［(50.10±1.23）% vs control, P<0.01］, incre ased S phase cell rate and induced the expression of CD44, but HA (100 μg/ml) i nhibited cell proliferation and the expression of CD44. Conclusion: HAase may degrade antiangiogenic HA of extracellular matrix, which may stim ulate proliferation of endothelial cells and enhance the curative effect of grow th factors to myocardial ischemia.