The habenula complex,an ancient and conservative nucleus during biological evolution, is involved in many i mportant biological functions,such as maternal behavior,pain,sleep,learning and reward.As an important node in the dopamine reward network,its functions(including the maintenance of individual survival while avoiding disadvantages)and mechanis m in the reward process have attracted wide attention.The dysfunction of habenula is closely related to many psychotic disorders,such as de-pression,schizophrenia and drug habituation.Habenula may also become a potential target for clinical treatment of these mental illnesses.So clarifying the role and neurobiological mechanisms of habenula in the central nervous system is of great theoretical and clinical value.

Objective:This study aimed to investigate the effect of human lymphatic endothelial cells (HLECs) on proteins secreted by epithelial ovarian cancer (EOC) cells SKOV3-pm4 with highly directional lymphatic metastasis. Methods:The supernatants of the four groups of cultured cells (A, SKOV3;B, SKOV3+HLEC;C, SKOV3-PM4;and D, SKOV3-PM4+HLEC) were collected. The proteins of these cells were detected by antibody arrays and iTRAQ-2D-LC-MALDI-TOF/TOF/MS. The screened significantly differential proteins were further analyzed by bioinformatics and validated in the human serum and cell culture medium by ELISA. Results:Progranulin (GRN) and vascular endothelial growth factor A (VEGF-A) were upregulated between groups C and A. In addition, insulin-like growth factor binding protein-7 (IGFBP-7) and secreted protein acid rich in cysteine (SPARC) were downregulated between groups D and C. Comprehensive bioinformatics analysis revealed that IGFBP7 interacted with VEGFA. VEGF exhibited the highest expression in ovarian cancer and IGFBP7 exhibited the lowest expression compared with the serum of the normal control group. Statistically significant differences were observed between the two substances. Conclusion:The HLEC microenvironment is closely associated with directional metastasis in lymph nodes with differential proteins, including matricellular proteins and adhesion factors. In particular, the upregulation of VEGFA and GRN and the downregulation of SPARC and IGFBP7 were closely associated with the directional metastasis of EOC cells in lymph nodes.

We wished to undertake molecular genetic typing and evaluate recombinants of the hepatitis-B virus (HBV) in Tibet (China). Multistage random sampling was used to collect HBsAg-positive samples. Nested polymerase chain reactions were used to amplify the whole sequence of the HBV. DNAstar, MEGA6 and SimPlot were used to assemble sequences, create phylogenetic trees, and undertake recombination analyses. Twelve whole sequences of the HBV of a Tibetan population were collected using these methods. Results showed that all 12 strains were C/D recombinants. Nine of the recombinations were at nt750, and the other three at nt1526. Therefore, the 12 strains could be divided into two types of recombinants: C/Da and C/Db. Analyses of the sequence of the whole genome revealed that the 12 strains belonged to genotype C, and that the nucleotide distance was > 4% between the 12 strains and sub-genotypes C1 to C15 in Genbank. The most likely sub-genotype was C1. Individuals with C/Da were from central and northern Tibet (e.g., Lasa, Linzhi, Ali) and those with C/Db recombinants were from Shannan in southern Tibet. These data suggest that the two types of recombinants had a good distribution in Tibet. Also, they can provide important information for studies on HBV recombination, gene features, virus evolution, as well as the control and prevention of HBV infection in Tibet.
Adult ; Female ; Genotype ; Hepatitis B ; virology ; Hepatitis B virus ; classification ; genetics ; isolation & purification ; Humans ; Male ; Phylogeny ; Recombination, Genetic ; Sequence Analysis, DNA ; Tibet ; Young Adult

We investigated the molecular characteristics of the full-length genome of 14 dengue serotype 1 virus (DENV-1)strains isolated in Sino-Myanmar border region in Yunnan Province,China during 2013-2015.Isolation of dengue virus was using C6/36 cell culture method.Viral RNA was extracted from virus isolates,and then the full-length genome was amplified by RT-PCR.The homology and phylogenetic analysis was made on the nucleotide and deduced amino acid sequences by bioinformatics software including ClastalX1.83 and MEGA6 etc.Results showed that fourteen strains of DENV-1 isolated from dengue fever cases,of these,9 strains from Ruili City of Dehong Prefecture,3 from Lincang Prefecture,2 from Kunming City.RT-PCR and sequencing indicated that the full-length genes (10 735 nt) of 14 DENV-1 strains were obtained,and their open reading frame (95-10 271) were coded 3 392 amino acid residues.The genotypes of DENV-1 were revealed by homology and phylogenetic analysis based on structural and non-structural proteins.Thirteen were genotype Ⅰ (G-Ⅰ) (7 from indigenous cases in Ruili and Lincang and 6 from imported case from Myanmar to Ruili,Lincang and Kunming),and 1 G-Ⅲ from imported case from India to Kunming.The phylogenic analysis indicated that the 13 isolates from Yunnan divided into 2 phylogenic subgroups,and they had a closer genetic relationship with the strains isolated from Southeast Asia.The gene sequences of the 13 G-Ⅰ strains have been acquired,the rate of their nucleotide homology and amino acid homology were 97.02 ％-100 ％ and 98.78 ％100 ％ respectively.Compared with 6 strains from Southeast Asia,nucleotide homology and amino acid homology were 96.53％-99.53％ and 97.33％-100％ respectively.Compared with prototype strain (US_Hawaii) of DENV-1,nucleotide homology and amino acid homology were 93.76％-94.45 ％ and 95.86 ％-96.91％ respectively.Compared with US_Hawaii strain,there were 44 and 150 different sites in amino acid of structural and non-structural proteins,respectively.The G-1 of DENV-1 have been popular in Sino-Myanmar border region in Yunnan,2013-2015.They have genetic diversity but multiple transmission sources were from Myanmar,and should strengthen control cross-border spread of dengue fever in this region.It is necessary to further study that change of the amino acid sites of Yunnan strains of DENV-1 is related to its antigenicity and pathogenicity.

Neuro-osteological interactions have an important role in the regulation of bone metabolism and regeneration. Neuropeptides combined with porous biphasic calcium phosphates (BCP) using protein adsorption may contribute to the acceleration of bone formation. In the present study, we investigated the effect of BCP combined with nerve growth factor (NGF) on the growth of osteoblasts in vitro and the combinational therapeutic effect on the repair of calvarial defects in vivo. NGF was separated and purified from Chinese cobra venom using a simplified three-step chromatography method. BCP combined with NGF exerted a potent effect on osteoblast differentiation, as evidenced by enhanced cell proliferation, increased ALP activity and the up-regulated expression of osteogenesis-related genes and proteins. Further, combinational therapy with BCP and NGF improved calvarial regeneration, which was superior to treatment with therapy alone, as observed using imageological and morphological examination and histological and immunohistochemical staining. The results confirmed the effect of neuro-osteological interactions through combinatorial treatment with NGF and BCP to promote osteogenesis and bone formation, which may provide an effective and economical strategy for clinical application.
Acceleration ; Adsorption ; Asian Continental Ancestry Group ; Calcium Phosphates ; Cell Proliferation ; Ceramics* ; Chromatography ; Cobra Venoms ; Elapidae* ; Humans ; In Vitro Techniques ; Metabolism ; Methods ; Nerve Growth Factor* ; Neuropeptides ; Osteoblasts ; Osteogenesis ; Regeneration ; Therapeutic Uses*

Objective:To investigate the correlations of laminin subunit gamma 3 (LAMC3) expression with prognosis of ovarian cancer (OC).Methods:LAMC3 protein expression was measured using immunohistochemical streptavidin-peroxidase-biotin connection method (IHC). Gene expression and related clinical data in the cancer genome atlas (TCGA) cohort and clinical proteomic tumor analysis consortium (CPTAC) were applied to analyse the correlation between gene and protein expressions and clinical outcomes. Correlations between LAMC3 and clinicopathological factors were evaluated using the Pearson χ2 test (2-sided). The probability of survival and significance was calculated using the Kaplan-Meier plot. The functional clustering of biological pathways enriched from co-expressed genes of LAMC3 was used to explore the possible mechanisms that LAMC3 might contribute to poor prognosis. Results:Based on the IHC results of 216 OC tissues or ovaries (including 208 tumors and 8 normal tissues) and 51 OC tissues (including 24 chemotherapy-resistant and 27 sensitive tissues), and the protein expression data from CPTAC (including 100 primary tumors and 25 normal tissues), the results showed that the protein expression of LAMC3 was significantly decreased in OC tissues compared with normal, decreased in advanced-stage tissues compared with early-stage tissues, and decreased in drug-resistant tissues compared with sensitive tissues (all P<0.05). Furthermore, low expression of LAMC3 protein was significantly associated with poor disease-free survival (DFS) and overall survival (OS) in 51 OC tissues ( P<0.01), consistent with the results that the low levels of LAMC3 mRNA predicted short DFS and OS in 489 OC tissues of the TCGA cohort ( P<0.05). The results suggested that low expression of LAMC3 might be the adverse factors for OC development, such as drug resistance and advanced tumors, and might be a risk indicator for prognosis. Moreover, functional clustering of biological pathways enriched from the co-expressed genes of LAMC3 in TCGA ovarian cohort indicated that LAMC3 potentially involved in regulation of OC via oncogene-pathways such as Ras associated protein 1 (Rap1), mitogen-activated protein kinase (MAPK), Ras and cell adhesion-related pathways such as extra cellular matrix (ECM)-receptor interaction and focal adhesion. It indicated that LAMC3 might contribute to short survival and tumor progression by regulation of the above pathways. Conclusion:Low expression of LAMC3 is related to poor prognosis and malignant progression in OC, and thus it is expected to be a new prognostic marker and therapeutic target for clinical treatment.

Objective: To investigate the possibility and diagnostic efficiency of ¹⁸F-NaF PET/CT bone scan after oral administration (PO) by comparing with that of intravenous injection (IV). Methods: Fifty patients (19 males, 31 females; average age: (52.8±11.7) years) with cancer who underwent PET/CT scans after oral and intravenous administration of ¹⁸F-NaF respectively with an interval of 2-7 d from June 2015 to September 2016 were prospectively enrolled. Single-phase ¹⁸F-NaF PET/CT was performed 60 min after IV, and dual-phase ¹⁸F-NaF PET/CT was performed 60 and 120 min after PO. All PET/CT images were reviewed, lesions were counted, and maximum standardized uptake value (SUV_max) and target/non-target (T/NT) ratios were calculated and compared. Paired t test was used. Results: Forty-one patients (15 males, 26 females; average age: (53.5±10.4) years) who finished all PET/CT scans were enrolled. The images at 120 min after PO was visually similar to the images at 60 min after IV. Three modalities detected the same cases and lesions (35 positive cases: 25 malignant, 8 benign, 2 cases with indefinite diagnosis; 302 lesions: 172 malignant, 108 benign, 22 ambiguous lesions). The SUV_{max-PO60 min} and SUV_{max-PO120 min} were lower than the SUV_{max-IV60 min} in the same lesion (18.22±12.64, 26.60±19.49 vs 28.07±16.34; t values: -12.36 and -3.59, both P<0.05). A total of 194 lesions were included for T/NT ratio analysis. T/NT_{IV60 min}, T/NT_{PO60 min} and T/NT_{PO120 min} were 2.76±1.30, 2.87±1.50, 2.98±1.42, respectively, and T/NT_{PO120 min} was higher than T/NT_{IV60 min} (t=3.18, P<0.05). Conclusion ¹⁸F-NaF PET/CT images after PO, especially at 120 min post-PO, has similar diagnostic power of lesion-detection and SUV_max-measurement with IV.

The identification of clinical questions for clinical practice guidelines of Chinese patent medicine （CPM） is important for subsequent evidence retrieval， evaluation of evidence quality， formation of recommendations. This paper described a methodological proposal for the identification of clinical questions for CPM guidelines to highlight the characteristics of Chinese patent medicine and reflect its effect in specific stage of the disease. Considering four aspects， namely， the drug of Chinese patent medicine （D）， the specific disease stage （S）， comparison （C）， and specific outcome （O）， DSCO framework has been proposed to formulate the clinical questions. Multi-source information through scientific research， policy or standard documents， and clinical data are suggested for collecting clinical questions， and clear selection criteria should be set to finalize the clinical questions to be addressed by the guideline. In addition， the above process needs to be transparently and publicly reported in order to ensure the clarity and completeness of the guidelines.

ObjectiveTo study the effect of organophosphate-solubilizing bacteria and compound bacteria on the photosynthesis and physiological and biochemical characteristics of leaves of Paris polyphylla var. yunnanensis， and to provide a reference for selecting suitable bacterial fertilizers in artificial cultivation of this medicinal species. MethodPot experiment was carried out indoor and the following groups were designed： control （CK）， inoculation with Bacillus mycoides （S1）， inoculation with B. wiedmannii （S2）， inoculation with B. proteolyticus （S3）， inoculation with B. mycoides and B. wiedmannii （S4）， inoculation with B. mycoides and B. proteolyticus （S5）， inoculation with B. wiedmannii and B. proteolyticus （S6）， and inoculation with B. mycoides， B. wiedmannii and B. proteolyticus （S7）. Then， the growth and development， photosynthesis， and various physiological and biochemical indexes of the leaves of this species were observed. ResultCompared with CK， the treatment groups showed decrease in content of malondialdehyde in the leaves （P<0.05）， particularly S7 （content was only about 1/3 that of the CK）. The leaf area， photosynthetic parameters， photosynthetic pigment content， soluble sugar content， soluble protein content， and activity of superoxide dismutase （SOD）， peroxidase （POD）， and catalase （CAT） in leaves of the treatment groups were all improved. Among them， the leaf area， soluble sugar content， and soluble protein content were the highest in S7， which were 2.8， 2.1， and 2.2 times that of the CK， respectively. SOD activity peaked in S6 （2.9 times higher than that in the CK） and the highest activity of POD and CAT was detected in S5 （1.5 times and 2.1 times， respectively higher than that in the CK）. ConclusionInoculation with different organophosphate-solubilizing bacteria or compound bacteria can promote the growth and development of P. polyphylla var. yunnanensis and improve its resistance to stresses. The combination of B. mycoides and B. proteolyticus and the combination of the three achieved the have the best effect. This study provides a reference for the selection of bacterial fertilizers for artificial cultivation of P. polyphylla var. yunnanensis.

The irrational use of Chinese patent medicines （CPM） is becoming more and more prominent， which makes the demand for clinical practice guidelines of CPM gradually increase. In order to make domestic scholars understand the latest developments and existing problems of the CPM guidelines， and promote its development， this paper introduced the concept of CPM guidelines， summarized the characteristics of the two development modes， namely “taking CPM as the key” and “taking disease/syndrome as the key”， and analyzed the current methodological status of developing and reporting CPM guidelines. Based on the existed problems， three suggestions have been put forward to optimize the quality of CPM guidelines， which were clarifying the target users and scope of CPM guidelines， establishing an open and transparent mechanism of the personnel involvement and process steps， and formulating implementable and operable recommendations for the use of CPM.