Animals ; Anti-HIV Agents ; pharmacology ; HIV ; drug effects ; genetics ; physiology ; HIV Infections ; drug therapy ; immunology ; virology ; Humans ; Mucous Membrane ; immunology ; virology ; Virus Replication ; drug effects

Objective To explore the effect and mechanism of L-arginine on fetal growth restriction by observing the expression of Bcl-2 and Bax in placenta.Methods Sixty patients with FGR were chosen,among which 30 cases who were treated with conventional ways were as convention group,and the other 30 cases who were treated with L-arginine were as L-arginine group.The birth weight and perinatal fetus outcome were detected.The central tissue of placenta got within 10 min after delivery were fixed by 100 g/L formaldehyde and embed by pa-raffin wax to observe the expression of Bcl-2 and Bax using immunohistochemistry.SPSS 11.5 software was used to analyze the data.Results Compared with convention group,the birth weight of L-arginine group was higher(P

Induced pluripotent stem cells (iPSCs) can be propagated indefinitely, while maintaining the capacity to differentiate into all cell types in the body except for the extra-embryonic tissues. This iPSC technology not only represents a new way to use individual-specific stem cells for regenerative medicine but also constitutes a novel method to obtain large numbers of disease-specific cells for biomedical research. However, the low efficiency of reprogramming and genomic integration of oncogenes and viral vectors limit the potential application of iPSCs. Chemical-induced reprogramming offers a novel approach to generating iPSCs. In this study, a new combination of small-molecule compounds (SMs) (sodium butyrate, A-83-01, CHIR99021, Y-27632) under conditions of transient folate deprivation was used to generate iPSC. It was found that transient folate deprivation combined with SMs was sufficient to permit reprogramming from mouse embryonic fibroblasts (MEFs) in the presence of transcription factors, Oct4 and Klf4, within 25 days, replacing Sox2 and c-Myc, and accelerated the generation of mouse iPSCs. The resulting cell lines resembled mouse embryonic stem (ES) cells with respect to proliferation rate, morphology, pluripotency-associated markers and gene expressions. Deprivation of folic acid, combined with treating MEFs with SMs, can improve the inducing efficiency of iPSCs and reduce their carcinogenicity and the use of exogenous reprogramming factors.

AIM: To confirm the differential expression genes in the rat ischemic brain. METHODS: The middle cerebral artery occlusion ischemic model was set up in rats. Fluorescence differential display reverse transcriptase-polymerase chain reaction (FDD RT-PCR) and reverse Northern blotting were used to fast confirm the differential expression genes. RESULTS: Nine differential expression sequence tags, including 6 known sequences and 3 unknown sequences, were confirmed. Among the known sequences, mus musculus ab1-interactor1,homo sapiens CGI-99 protein, tissue inhibitor of metalloproteinase 3 and homosapiens nuclear receptor co-repressor were up-regulated while homo-sapiens nuclear matrix protein p84 and coatomer protein complex, subunit gamma 2 were down-regulated. CONCLUSIONS: ① Combination of fluorescence differential display reverse transcriptase-polymerase chain reaction (FDD RT-PCR) and reverse Northern blotting is a method to fast-confirm the differential expression genes; ② There are differential expression genes in ischemic brain regions compared to non-ischemic parts. [

AIM To construct a non-normalized cDNA library from Agkistrodon acutus venom gland as an imtial step to develop new and more effective venom by genetic engineering technique for screening and expressing target genes. METHODS The total RNA was extracted from fresh venom gland using Trizol. mRNA was reversely transcripted to cDNA using superscriptⅡ reverse transcriptase. Second-strand synthesis was performed using DNA polymeraseⅠ. After adding EcoRⅠ adaptor, phosphorylating the end and digesting with XhoⅠ, the cDNA was collected in five fractions (<0.25 kb, 0.25-0.5 kb, 0.5-1 kb, 1-2 kb and >2 kb) using the QIAquick Gel Extraction kit and ligated to pBluescriptⅡ vectors. The five libraries obtained were plated by infecting E.coli DH10B, constructing a cDNA library of Agkistrodon acutus venom gland. Sequencing clones at random, 8696 high quality 5′ end expressed sequenced tags (ESTs) were obtained and analyzed. The initial sequences were assembled into 2855 clusters. Among which, one of the clusters (Agkihagin) consisting of 74 ESTs was identified as a novel metalloprtoteinase based on RT-PCR and sequence analysis. RESULTSThe titers of library were 2.048×106. The novel metalloproteinase belonged to PⅢ type metalloproteinase. Its open reading frame was composed of 1827 nucleotides and coded a pre-zymogen of 608 amino acid with zinc-binding domain for metalloproteinase and Asp-Glu-Cys-Asp(DECD) domain for disintegrin. CONCLUSION The capacity of cDNA library of venom gland is above the general level of cDNA library. It would be a helpful platform to construct a catalog for transcripts in the venom gland of the Agkistrodon acutus. The sequence analysis indicates that the deduced amino acid sequence of the identified gene for metalloproteinase share the highest 87% identity with the metalloproteinase genes of other snakes in the GenBank. It lays a good foundation for the study of structure-function relationships of snake venom metalloproteinases.

In order to provide scientific basics for exploitation and sufficient application of Dendrobium officinale leaves resources, the phenol-sulfuric acid method was applied to determine the polysaccharide content. The monosaccharides were derivated by PMP and the derivatives were identified by HPLC-DAD-ESI-MS(n) and the contents of mannose and glucose were determined simultaneously. Similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine (2004A) was employed to generate the mean chromatogram and similarity analysis of the samples was carried out. The results demonstrated that polysaccharide content, monosaccharide compositions and composition ratio had an obvious difference between stems and leaves. The polysaccharide content of stems was higher than that of leaves. Monosaccharide composition in leaf was significantly different from that in stem. The polysaccharide from stems was composed of mannose and glucose, however the polysaccharide of leaves was acid heteropolysaccharide and was mainly composed of five monosaccharides, including mannose, galacturonic acid, glucose, galactose and arabinose. The similarity value of the 14 batches was above 0.9, indicating that similarity of fingerprints among different samples was high. The study can provide evidence for expanding the medicinal parts of D. officinale.
Chromatography, High Pressure Liquid ; Dendrobium ; chemistry ; Mass Spectrometry ; Plant Extracts ; chemistry ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Polysaccharides ; chemistry

An 81-year-old male presented with an 8-year history of recurrent ulcer on the left dorsal foot which gradually spread to involve both lower limbs. Physical examination revealed no abnormality of any organ systems and no palpable superficial lymph nodes. Skin examination showed erythematous swelling of the left dorsal foot with an ulcer sized 7 cm × 10 cm on the surface. Tendon was visible at the base of the ulcer, and the ulcer margin was elevated giving a dyke-like appearance. The perilesional skin was purple-brown. There were several millet-like papuloid lesions circularly arranged at the inner side of the right foot as well as dark erythematous or brown nodules and pigmented patches with tenderness on both lower limbs. Histopathology of the ulcer of the left dorsal foot and papuloid lesions on the right foot revealed a visible epidermotropic infiltrate in the epidermis as well as an infiltration throughout the entire dermis with medium-sized atypical lymphoid cells with obvious mitoses. Immunohistochemical examination showed the coexpression of both T cell markers (including CD3, CD45RO, CD43) and B cell marker (CD20), with scatted positive staining for PAX-5and negative staining for CD79α or CD1 9. PCR confirmed the rearrangement of T cell receptor (TCR)-γgene. A diagnosis of peripheral T cell lymphoma unspecified was made in view of the rearrangement of TCR-γgene and above findings. The patient was treated with the following modified CHOP regimen: intravenous cyclophosphamide 0.8 g, leurocristine 2 mg and epirubicin hydrochloride 60 mg, as well as oral prednisone 15 mg twice daily for 5 days every 3 weeks (one treatment session). After 3 treatment sessions, the lesions improved markedly.

【Objective】To study the effect of the specific p38 mitogen-activated protein kinase(p38 MAPK) inhibitor SB203580 on apoptosis of cerebellar granule neurons induced by low potassium.【Methods】Apoptosis was induced by switching the cultured cerebellar granule neurons from a culture medium containing K+ 25 mmol*L-1 to a medium containing K+ 5 mmol*L-1 (cLK).Fragmentation of DNA was analyzed using agarose gel eletrophoresis.SAPK/JNK activity was measured by SAPK/JNK assay kit.【Results】Low potassium resulted in apoptosis as characterized by morphological and biochemical features,but the specific p38 MAPK inhibitor SB203580 improved the survival of cerebellar granule neurons cultured in cLK medium by blocking apoptosis in a concentration-dependent manner.The expression and phosphorylation of c-Jun increased and the activity of c-Jun N-terminal protein kinase (JNK) elevated when cerebellar granule neurons were cultured in cLK medium.But when the cerebellar granule neurons cultured in cLK medium were exposed to 25 μmol*L-1 SB203580,the expression and phosphorylation of c-Jun and the activity of JNK were both decreased evidently.【Conclusions】These results indicate that SB203580 inhibits the activation of JNK and phosphorylation of c-Jun,and therefore protects granule neurons from apoptosis induced by low potassium.

AIM To investigate the protection of plicamycin on apoptosis in cerebellar granule neurons (CGN) of rat. METHODS TUNEL, Hoechst 33258 staining, agarose gel electrophoresis and fluorescein diacetate staining were used to detect morphological and biochemical characteristics of apoptosis in primary rat CGN. RESULTS Being pre-incubated with plicamycin for 1 h and lasting for 24 h, rat CGN apoptosis induced by low potassium basal modified Eagle′s medium for 24 h was inhibited in a plicamycin concentration-dependent manner. This effective concentrations of plicamycin were from 50 to 200 nmol·L-1, and the maximum inhibitory rate of plicamycin on CGN apoptosis was near 80% at 200 nmol·L-1. CONCLUSIONPlicamycin inhibits rat CGN apoptosis induced by low potassium.

Objective To investigate the clinical features, effective treatment, survival and prognostic factors of second primary tongue squamous cell carcinoma (SPTSCC) after nasopharyngeal carcinoma (NPC) radiotherapy. Methods The clinical data of 35 cases with SPTSCC after NPC radiotherapy were analyzed retrospectively. Kaplan-Meier method, Log-Rank test and COX proportional hazard mode was performed for statistical analysis. Results 3-year and 5-year overall survival rates were 55 % and 47 %, respectively, lymph node metastasis rate was 5.71 %. Univariate analysis indicated that gender (χ2 = 8.89, P = 0.00), T classification (χ2= 5.58, P= 0.02), clinical stage (χ2 = 8.51, P= 0.04) and treatment methods (χ2 = 29.37, P = 0.00) were important factors of prognosis. Multivariate analysis showed that treatment methods (P = 0.00) and T classification (P = 0.03) were independent prognostic factors. Operative treatment group had better prognosis than the non-operative treatment group, the difference was statistically significant (P <0.05), male patients in the risk of SPTSCC was higher than the female patients, and the incidence of SPTSCC was increased along with extension of the time after NPC radiotherapy. Conclusion The rate of the lymph node metastasis is lower for SPTSCC after NPC radiotherapy and treatment patterns and T stage are independent prognostic factors. Long-term follow-up after NPC radiotherapy is necessary to the early diagnosis of SPTSCC, so that to give surgery or combined therapy with surgery in order to achieve a good effect.