Objective To observe the effect of intercellular adhesion molecule on neuronal differentiation of P19 cells in vitro.Methods Monolayer P19 cells were cultured on the plates coated with 0.1% gelatin, 2.5 mg/L and 10 mg/L N-cad-Fc (a fusion protein of the N-cadherin extracellular domain and the IgG Fc domain), and then 5 × 10-7mol/L retinoic acid (RA) was used to induce neuronal differentiation of P19 cells. Neuron-specific class Ⅲβ-tubulin (Tuj-1) antibody was examined by immunofluorescence staining. The protein expressions of oct3/4 in stem cells, nestin in precursor cells, ngnl and map2 in neurons and gfap in glial cells were detected by reverse transcription-polymerase chain reaction (RT-PCR).Results RA at 5 × 10-7mol/L could induce P19 cells to differentiate into neurons. The average differentiation rates on 0. 1% gelatin, 2. 5 mg/L and 10 mg/L N-cad-Fc were 7. 8%, 28. 6% and 26.8%, respectively. Cells cultured on N-cad-Fc coated plates dramatically up-regulated the expressions of neural-specific proteins ngnl and map2. The expression of stem cell specific protein oct3/4 was decreased ahead of schedule and the expression of glial cell specific protein gfap could not be found by RT-PCR. Conclusions RA can induce P19 cells to differentiate into neuronal cells. Intercellular adhesion molecule has promotional effect on cell differentiation.

OBJECTIVE: To identify Pogostemon cablin and Cortex Magnoliae Officinalis in Huoxiang zhengqi pills by Gas Chromatography-Mass Spectrometry. METHODS:Pogostemon cablin and Cortex Magnoliae Officinalis in Huoxiang zhengqi pills were extracted by ultrasound extraction technology and identified on SPB-1701 column(30 m?0.25 mm?0.25 ?m) with He as carrier gas and EI as ion source at an electron energy of 70 eV. RESULT:The main components such as Patchouli alcohol from Pogostemon cablin,and Magnolol and Honokiol from Cortex Magnoliae Officinalis were identified simultaneouly. CONCLUSION: The method is simple,rapid,accurate and reliable,and applicable for the quality control of Huoxiang zhengqi pills.

Objective To investigate the sensitivity of tumor cells to photodynamic therapy(PDT), and to select the optimal photo-dose and photo density for PDT to the cultured cells. Methods Cells of SHEE and SHEEC cell lines were divided into 21 groups randomly,after 24 h inoculation, the cells accreted on paries of culture capsule completely, we replaced M199 complete culture solution with 30 μg/rnl Photofrin-Ⅱ solution 100 μl and then replacd with M199 complete culture solution without Photofiin-Ⅱ after 150 min of incubation in Photofrin-Ⅱ. Within 15 min,we dealed the cells with PDT using Photofrin-Diomed 630 under three different power densities:25 raW/cm~2,50 mW/cm~2 and 100 mW/cm~2 for 10 s,20 9,30 s,50 s, 100 s, 150 s and 200 s respectively,and then continue for 24 h culture. We examined the inhibiting effect on cell line SHEE and SHEEC under PDT by the method of CCK-8. Results There was no significant difference in the inhibition ratio with same power density of PDT between cell line SHEE and SHEEC under the concentration of 30 μg/ml Photofrin-Ⅱ. However, the inhibition ratio increased with the raising of photo-dose of PDT,but there was a platform stage after the 2.5～30 J/cm~ 2 photo-dose. Conclu-sion The difference for photodynamic sensitivity between human immortalization esophageal epithelial cell line SHEE and the malignant transformation cell line SHEEC is not significant. The targeting of PDT to malignant tumor cell may not be involved in the photosensitivity for tumor cell.

ObjectiveTo identify the curative effect of minimally invasive plate osteosynthesis (MIPO) and humeral head replacement in treatment of elderly patients with Neer four-part fractures.MethodsUsing the deltoid pectoral approach,28 patients with fresh Neer four-part fractures were treated by the locking plate combined with MIPO (Group A) and 27 by the humeral head replacement (Group B).Neer score,Constant-Murley score and simple shoulder test (SST) questionnaire were adopted for assessing the treatment outcome.ResultsGroup A was followed up for mean 32.1 months,which showed screws protruding into the joint space in two patients,tuberosity upward and backward displacement in one and femoral head ischemic necrosis in one.The mean visual analog scale (VAS) score,the mean Neer score and the mean Constant-Murley score were 2.2 points,88.6 points and 86.5 points respectively.There were average 9.0 answers for “yes” in the SST questionnaire.Group B were followed up for mean 34.6 months,which showed shoulder dislocation or subluxation in four patients,tuberosity displacement or excessive reduction in eight.The mean VAS score,the mean Neer score and the mean Constant-Murley score were 2.4 points,78.9 points and 77.3 points respectively.The mean number of answer for “yes” in SST questionnaire was 8.0 questions.There showed no statistical difference in VAS score between the two groups.While statistical difference was found in complications,Neer score,Constant-Murley score and SST score between two groups,with Group A superior to Group B.Conclusions For most elderly patients with Neer four-part fractures,MIPO has satisfactory results under strict control of surgical indications and technical tips.The humeral head replacement surgery still has many unresolved problems and needs careful consideration.

Object To demonstrate the naked truth of four therapeutic features of TCM (cold, hot, warm and cool) from biophysical and biochemical point of view. Methods Growth thermogenesis curves of Escherichia coli were determined under different conditions by microcalorimetry to give biothermodynamic parameters, such as multiplication constant (k), median inhibitory concentration (IC 50 ), and enthalpy (?H). Differences between folia ginseng and flos ginseng were comprehensively compared with reference to classical Chinese literatures on material medica. Results The bacteriostatic curve of both drugs were basically similar. With increasing concentration of decoctions of both drugs, the lag phase was delayed and the peaks shifted to the right. While the difference of ?H was stable, which decreases with the increasing concentration of Folium ginseng, but increases with increasing concentration of Flos ginseng. Conclusion Microcalorimetry is not only a new effective method to describe the therapeutic features of TCM, but also could be used as a tool for the screening of active TCM.

Objective To investigate the relationship between expression of DAS-1 in gastric cardia intestinal metaplasia(CIM)and gastric cardia adenocarcinoma (GCA). Methods The cancerous tissues and CIM tissues (2 cm apart from caneer) obtained from 65 patients with GCA were examined for the expression of DAS-1 protein using immunohistoehemistry. The CIM tissues (<2 cm below Z line) obtained from 15 outpatients and inflammatory mucosa from 25 outpatients were also examined for expression of DAS-1 protein. Results The type Ⅲ IM was accounted for 55.4% (36/65) in GCA patients, which was significantly higher than that in outpatients [13.3% (2/15), P<0.01]. The positive rate of DAS-1 expression in cancerous tissues [78.5 % (51/65)] was also significantly higher than that in CIM tissues [38.8 %(30/80), P<0.01]. The expression of DAS-1 protein in IM tissues was gradually increased from type Ⅰ (0%) to type Ⅲ (71.1%) with positive correlation (P<0.01). Conclusions The type Ⅲ IM with over-expression of DAS-1 is closely related to GCA, which might be one of important precancerous lesions for GCA.

OBJECTIVE: To choose the disease-causing gene in a Chinese pedigree with ankylosing spondylitis (AS) by whole-exome sequencing (WES), and provide theory basis for mechanism of disease. METHODS: Clinical data of AS pedigree were collected, including 2 males, the age were 48 and 18 years old, the course of disease were 23 and 4 years. Whole blood genomic DNA of AS was extracted to perform whole exome sequencing, the results were compared with human databases, common variations which had been reported were wiped out, then non synonymous single nucleotide variants(SNVs) from the family members were combined, and candidate genes was selected initially. RESULTS: Totally 80 G data was obtained from AS family with high quality.By comparing results between patient and normal subject, and filtering with number of biological database, the result showed heterozygous mutation of JAK2 gene 12 exon c.1709 A>G (p.Tyr570Cys) may be the potential disease-causing gene. The variant c.1151T>C of MUC3A gene may be one of the causes of intestinal symptoms in the family members. CONCLUSION It is feasible to find t candidate gene mutations of AS by Exon sequencing. The mutation c.1709 A>G in gene JAK2 identified by whole exome sequencing might be the pathogenic mutation in this AS pedigree.
Exome ; Humans ; Male ; Mucin-3 ; Mutation ; Pedigree ; Spondylitis, Ankylosing ; Whole Exome Sequencing

Objective:To explore the clinical observation of Xiaozhang Plaster for external use in the treatment of postprandial discomfort syndrome of qi stagnation functional dyspepsia.Methods:Randomized controlled trial. From January 2019 to January 2022, 128 patients with postprandial discomfort syndrome of functional dyspepsia of qi stagnation type in our hospital were selected and divided into two groups by random number table, with 64 cases in each group. Both groups were orally treated with mosapride citrate tablets. On this basis, the treatment group was combined with external use of Xiaozhang Plaster, and the control group was combined with placebo patch. Both groups were treated for 2 weeks. Traditional Chinese Medicine syndrome score, symptom severity and attack frequency score were performed before and after treatment. The quality of life of the patients was assessed with SF-36 Short-form health survey-36 (SF-36), anxiety and depression were assessed with Hospital Anxiety and Depression Scale (HADS), and the growth hormone releasing peptide (Ghrelin), calcitonin gene-related peptide (CGRP) The levels of gastrin 17 (G-17), pepsinogen I (PG I) and pepsinogen Ⅱ (PG Ⅱ) levels were observed by ELISA, and clinical efficacy was evaluated.Results:The total effective rate was 96.88% (62/64) in the treatment group and 81.25% (52/64) in the control group. The difference between the two groups was statistically significant ( χ2=8.02, P<0.05). After treatment, the score and total score of epigastric distension, epigastric distension and pain, fullness of both flanks, loss of appetite in the treatment group were significantly lower than those in the control group ( t=38.43, 32.39, 38.43, 32.87, 33.74, P<0.01), the score of symptom severity and attack frequency were significantly lower than those in the control group ( t=61.42, 33.46, P<0.01), and the score of SF-36 was significantly higher than that in the control group ( t=8.26, P<0.01); The score of HADS was significantly lower than that of the control group ( t=38.06, P<0.01). After treatment, the levels of CGRP and G-17 in the treatment group were significantly lower than those in the control group ( t values were 6.22, 29.51, P<0.01), the level of Ghrelin, and the levels of PG Ⅰ, PG Ⅱ were significantly higher than those in the control group ( t=23.85, 13.26, 19.53, P<0.01). Conclusion:On the basis of basic drug treatment, the application of Xiaozhang Plaster for external use to intervene patients with postprandial discomfort syndrome of qi stagnation functional dyspepsia can improve the quality of life, gastrointestinal hormones, anxiety and depression symptoms, and the effect is remarkable.

OBJECTIVETo study the inhibitory effects of Taxus chinensis var. mairei Aqueous Extract (TAE) on SGC-7901 and MCF-7 cells, and to explore its mechanisms.

METHODSThe inhibitory effects of TAT and Paclitaxel on the proliferation of SGC-7901 and MCF-7 cells were tested by MTT method. Their effects on the morphology of SGC-7901 and MCF-7 cells were observed by microscope. Its effects on the cell apoptosis were detected by flow cytometry.

RESULTSThe TAE had inhibitory effects on the proliferation of tumor cells, and its mechanisms were correlated to inducing the apoptosis of tumor cells.

CONCLUSIONTAE had inhibitory effects on the proliferation of tumor cells.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; MCF-7 Cells ; Plant Extracts ; pharmacology ; Taxus ; chemistry

Objective To construct the molecular transmission networks of HIV-1 CRF01 AE strains circulating in China and to analyze their characteristics. Methods Sequences of the pol genes of Chinese CRF01 AE strains were downloaded from Los Alamos database and the related demographic informa-tion was checked. Transmission networks were created by four steps,including phylogenetic tree construc-tion,transmission cluster extraction,minimum genetic distance identification and network visualization. Chi-square test was performed to analyze the differences in the distribution of different populations in the networks and the differences in the distribution of subjects with different degrees among different sub-populations. De-scriptive analysis was used to investigate the transmission links between sub-populations and various regions. Results The 2 419 sequences constituted 250 separate networks including 847 nodes and 610 edges. The number of subjects covered by different networks ranged from 2 to 25. Subjects with degree ≥2 represented only 26. 4%(224 / 847)of network-individuals,but were connected with 66. 5%(563 / 847)of all network-individuals. There were 37. 6%(669 / 1 781)of men who have sex with men(MSM),28. 4%(102 / 359) of heterosexual men,16. 1%(10 / 62)of intravenous drug users(IDUs)and 30. 4%(66 / 217)of hetero-sexual women involved in the networks(χ2 =23. 774,P﹤0. 001). The percentage of subjects with degree ≥2 was high in MSM(10. 8% ,193 / 1 781)compared to that in heterosexual men(5. 6% ,20 / 359)(P =0. 002)and heterosexual women(4. 6% ,10 / 217)(P=0. 004). Of the 669 MSM in the networks,95. 5%(639 / 669)linked to other MSM and only 2. 4% (16 / 669)linked to heterosexual women. However, 15. 1%(10 / 66)of the heterosexual women in the networks linked to MSM. Of the heterosexual men in the networks,35. 3%(36 / 102)linked to heterosexual women,9. 8%(10 / 102)linked to MSM. Of the sub-jects in the networks,20. 9%(177 / 847)linked to other regions' individuals. Conclusion The super-spreaders played an important role in the molecular transmission networks of HIV-1 CRF01 AE strains even though they were in a minority. The transmission of HIV-1 CRF01 AE strains between sub-populations and various regions was complicated and active.