Establishment of a reporter system for estimating activation of human hepatic stellate cells based on COL1A1 promoter and enhanced green fluorescent protein / 北京大学学报(医学版)

OBJECTIVE@#To establish a visual reporting system for evaluating the activity of collagen Ⅰ α 1 chain (COL1A1) gene promoter in immortalized human hepatic stellate cells, so as to estimate the activation status of the cells and provide a new cell model for the screening and study of anti-hepatic fibrosis drugs.@*METHODS@#The promoter sequence of human COL1A1 was amplified from the genomic DNA of human hepatocarcinoma cell line HepG2. Based on the pLVX-AcGFP1-N1 plasmid, the recombinant plasmid pLVX-COL1A1-enhanced green fluorescent protein (EGFP) was constructed, in which the enhanced green fluorescent protein gene expression was regulated by the COL1A1 promoter. The monoclonal cell line was acquired by stably transfecting pLVX-COL1A1-EGFP into the immortalized human hepatic stellate cell line LX-2 by the lentivirus packaging system and screening. The cell line was treated with transforming growth factor-β1 (TGF-β1) or co-treated with TGF-β1 and drugs with potential anti-hepatic fibrosis effects. The EGFP fluorescence intensity in cells was analyzed by the fluorescence microscope and ImageJ 1.49 software using a semi-quantitative method. The COL1A1 and EGFP mRNA were detected by reverse transcription real-time quantitative PCR (RT-qPCR), and corresponding proteins were detected by Western blot.@*RESULTS@#The recombinant plasmid pLVX-COL1A1-EGFP with the expression of EGFP regulated by COL1A1 promoter was successfully constructed. Kozak sequence was added to enhance the expression of EGFP, which was identified by double digestion and sequencing. The LX-2 monoclonal cell line LX-2-CE stably transfected with pLVX-COL1A1-EGFP was obtained. After co-treatment with TGF-β1 and 5 μmol/L dihydrotanshinone Ⅰ with potential anti-hepatic fibrosis effect for 24 h, the total fluorescence intensity and the average fluorescence intensity of LX-2-CE were lower than those in TGF-β1 single treatment group (P < 0.05), the intracellular mRNA and protein levels of COL1A1 and EGFP were also lower than those in the TGF-β1 single treatment group (P < 0.05).@*CONCLUSION@#A reporter system for estimating activation of hepatic stellate cells based on COL1A1 promoter regulated EGFP expression is successfully constructed, which could visually report the changes in COL1A1 expression, one of the activation-related markers of hepatic stellate cells, in vitro. It provides a new cell model for the screening and study of anti-hepatic fibrosis drugs.

Epidemiological distribution of genotypes and sub-genotypes of hepatitis B virus in 15 ethnic groups in China / 中华流行病学杂志

Objective: To understand the distribution of genotypes and sub-genotypes of HBV in different ethnic groups in China. Methods: The HBsAg positive samples were selected by stratified multi-stage cluster sampling from the sample base of national HBV sero-epidemiological survey in 2020 for the amplification of S gene of HBV by nested PCR. A phylogeny tree was constructed to determine the genotypes and sub-genotypes of HBV. The distribution of genotypes and sub-genotypes of HBV were analyzed comprehensively by using laboratory data and demographic data. Results: A total of 1 539 positive samples from 15 ethnic groups were successfully amplified and analyzed, and 5 genotypes (B, C, D, I and C/D) were detected. The proportion of genotype B was higher in ethnic group of Han (74.52%, 623/836), Zhuang (49.28%, 34/69), Yi (53.19%, 25/47), Miao (94.12%, 32/34), Buyi (81.48%, 22/27). The proportions of genotype C were higher in ethnic groups of Yao (70.91%, 39/55). Genotype D was the predominant genotype in Uygur (83.78%, 31/37). Genotype C/D were detected in Tibetan (92.35%,326/353). In this study, 11 cases of genotype I were detected, 8 of which were distributed in Zhuang nationality. Except for Tibetan, sub-genotype B2 accounted for more than 80.00% in genotype B in all ethnic groups. The proportions of sub-genotype C2 were higher in 8 ethnic groups, i.e. Han, Tibetan, Yi, Uygur, Mongolian, Manchu, Hui and Miao. The proportions of sub-genotype C5 were higher in ethnic groups of Zhuang (55.56%, 15/27) and Yao (84.62%, 33/39). For genotype D, sub-genotype D3 was detected in Yi ethnic group and sub-genotype D1 was detected in both Uygur and Kazak. The proportions of sub-genotype C/D1 and C/D2 in Tibetan were 43.06% (152/353) and 49.29% (174/353). For all the 11 cases of genotype I infection, only sub-genotype I1 was detected. Conclusions: Five genotypes and 15 sub-genotypes of HBV were found in 15 ethnic groups. There were significant differences in the distribution of genotypes and sub-genotypes of HBV among different ethnic groups.

Mechanism of Anti-tuberculosis Particles （Kanglao Granule） Based on Network Pharmacology and Experimental Research / 中国实验方剂学杂志

ObjectiveTo explore the potential anti-tuberculosis mechanism of Kanglao granule through network pharmacology. MethodThe active components of Kanglao granule were retrieved from related databases and the potential targets of the components from SwissTargetPrediction. Targets of the tuberculosis were screened from GeneCards and National Center for Biotechnology Information （NCBI）， and the anti-tuberculosis targets of the prescription were further identified. STRING and Cytoscape 3.8.0 were employed to construct the Chinese medicinal-disease target-signaling pathway network and screen core targets. Then gene ontology （GO） term enrichment and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment were performed. Finally， AutoDock Vina was used for molecular docking between the active components of the prescription and key proteins and Western blotting for verifying the interaction between them. ResultA total of 29 important chemical components in the prescription were screened out， including β-sitosterol， sesamin， and kaempferol. A total of 28 key anti-tuberculosis targets were retrieved， such as protein kinase B1 （Akt1）， epidermal growth factor receptor （EGFR）， hypoxia inducible factor-1A （HIF-1A）， proto-oncogene tyrosine-protein kinase （SRC）， and matrix metalloproteinase-9 （MMP-9）. Bioinformatics analysis showed the 28 targets were involved in 41 GO terms such as oxygen metabolism， nucleic acid transcription， and metabolic enzyme pathway， and 28 key KEGG pathways， including Mycobacterium tuberculosis signaling pathway and phosphatidylinositol 3 kinase/protein kinase B pathway. Molecular docking results showed that Akt1 had the strongest binding affinity to sesamin. In vitro experiment indicated that sesamin inhibited the growth of M. tuberculosis by suppressing the phosphorylation of Akt1. ConclusionKanglao granule improved the sterilization level and immune response through multi-component， multi-target， and multi-pathway interactions， thereby achieving therapeutic effect on tuberculosis. Akt1 is one of the important targets involved in the treatment of tuberculosis.

Mechanism of Zhigancao Tang Against Coronary Heart Disease： An Exploration Based on Thought of "Four in One" / 中国实验方剂学杂志

"Four in one" is a research idea of identifying the classic prescriptions from the following four dimensions： "nature， location， tendency， and syndrome". The multi-dimensional analysis of the mechanism of classic prescription Zhigancao Tang in treating coronary heart disease helps to understand the syndrome differentiation and treatment thoughts of ZHANG Zhong-jing. The coronary heart disease results from deficiency. The efficacy of Zhigancao Tang in treating coronary heart disease can be elucidated from the "nature，location，tendency， and syndrome". In terms of nature， Zhigancao Tang is pungent and sweet in flavor and warm in property， with the sweet responsible for tonifying deficiency， the pungent for dispersing Yang， resolving Yin， and eliminating surplus pathogen， and the warm for moving Yangqi， nourishing blood， and promoting blood circulation. In terms of location， Zhigancao Tang mainly acts on vessels for restoring the normal circulation of blood in the vessels and improving coronary artery stenosis and the resulting ischemia and anoxia. In terms of tendency， Zhigancao Tang tends to affect the upper and inner parts of the body to tonify deficiency in Zangfu organs， promote fluid production， nourish nutrient blood， and dissipate cold simultaneously， thus alleviating chest impediment. In terms of syndrome， Zhigancao Tang is applicable to fluid exhaustion with blood dryness and Yin-yang-qi-blood deficiency syndrome， manifested as regularly or irregularly intermittent pulse and severe palpitation. Zhigancao Tang has been widely used for the treatment of over 70 diseases classified into 10 systems， especially the cardiovascular diseases， in traditional Chinese medicine （TCM） and western medicine. As the “one” of “four in one”， Zhigancao Tang is composed of multiple Chinese herbs and its therapeutic effect is superior to the sum of its parts. It ameliorates the coronary heart disease by resisting inflammation， protecting against ischemia-reperfusion injury， adjusting the ion channels of myocardial cells， and participating in atrial remodeling and hematopoiesis. Its mechanism and clinical efficacy in the treatment of coronary heart disease have been verified by clinical and experimental studies. The utilization of the thought of "four in one" to analyze classical prescriptions enables the combination of prescriptions with syndromes， which is of great significance to the clinical application and modern development of classical prescriptions.

Progress on Molecular Mechanism of Curcumin and Its Nano-preparation Against Liver Cancer / 中国实验方剂学杂志

There are certain limitations in the application of liver transplantation， resection and radiofrequency ablation for liver cancer. Therefore，specific and selective new drugs are needed to provide better treatment. Curcumin is a hydrophobic polyphenol with a wide range of activities，such as anti-inflammatory，antibacterial，anti-oxidant and anti-tumor properties. Its nano-preparation has stronger growth inhibition and pro-apoptosis effects on tumor cells. Literature retrieval found that curcumin's anti-liver cancer molecular mechanisms include inhibiting cell proliferation by regulating the expressions of relevant miR，glyoxalase 1（GLO1），CD133 and vascular endothelial growth factor （VEGF），inhibiting signal transducer and activator of transcription （STAT3） and YAP expression to induce cell apoptosis，regulating the heat shock protein 70 （HSP70）-Toll-like receptor 4 （TLR4） signaling pathway，Wnt/β-catenin and transforming growth factor（TGF）/epithelial-mesenchymal transition（EMT） pathways，nuclear factor-κB （NF-κB） signaling pathway and nuclear factor E2-related factor 2（Nrf2）/ Kelch-like ECH-related protein 1（Keap1） signaling pathway，inhibiting p38 mitogen-activated protein kinase （MAPK） phosphorylation，to reduce Lin28B expression，regulating phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway and inhibiting G protein-coupled receptors （GPR81）/hydroxycarboxylic acid receptor-1 （HCAR-1） expression to reverse transformation therapy resistance. Curcumin nano-preparation mainly includes polymer micelles，liposomes，loaded microbubbles，nanocapsules and nanoparticles. Curcumin is mainly delivered to liver cancer cells to rapidly release the drug，enhance the anti-liver cancer effect and reduce toxic and side effects in normal liver cells. The mechanisms include activation of DR5/Caspase-mediated exogenous apoptosis pathway and VEGF/VEGF receptors （VEGFRs） signaling pathway，loss of mitochondrial membrane potential and increase of intracellular reactive oxygen species （ROS）. This paper summarizes the molecular mechanism of curcumin and its nano-preparation against liver cancer，in order to further define the molecular mechanism of liver cancer and provide a new reference for its clinical diagnosis and treatment.

The prognosis of hepatitis E superinfection in hospitalized chronic hepatitis B patients in China: A systematic review and Meta-analysis / 中华疾病控制杂志

@#Objective To evaluate the prevalence and prognosis of hepatitis E virus ( HEV) su- perinfection in patients with chronic hepatitis B ( CHB) ． Methods Electronic databases of Pubmed， CNKI，and Wanfang were searched，and references of the relevant articles for literatures associated with HEV superinfection in CHB patients in China published during 2008－2019 were screened． Studies which met the inclusion and exclusion criterias were selected and related data were pooled for analysis． The Ｒ 3. 5. 2 and Ｒevman 5. 3 were used to measure heterogeneity with the I2 statistic and pooled incidence and odds ratio ( OＲ) ． Ｒesults 28 literatures were eligible for inclusion． The results showed the rate of HEV superinfection in hospitalized CHB ( CHB-HEV) patients was 13. 6% ( 95%CI: 12. 5% －14. 9%) ． The pooled incidence of liver failure in CHB-HEV patients was 34. 7% ( 95%CI: 29. 6%－40. 1%) ，which was higher than CHB patients ( OＲ= 10. 9，95%CI: 6. 8－17. 4) ，CHB patients superinfected with hepatitis A virus ( CHB-HAV) ( OＲ= 3. 8，95%CI: 2. 4－6. 2) ，and hepatitis E patients ( OＲ= 5. 1，95%CI: 3. 8－ 6. 8) ． The pooled mortality of CHB-HEV patients was 13. 8% ( 95%CI: 10. 3－17. 8) ，which was higher than CHB patients ( OＲ = 8. 5，95%CI: 4. 0－18. 3) ，CHB-HAV patients ( OＲ = 14. 4，95%CI: 4. 8－ 42. 9) ，and hepatitis E patients ( OＲ= 4. 4，95%CI: 3. 0－6. 4) ． Conclusion In China，HEV sueperinfection on chronic hepatitis B patients are common，and is an important cause of liver failure and death．

Genetic analysis of mitochondrial pcox1 and ribosomal 18S rRNA genes in Eurytrema pancreaticum isolates from goats in Huaihua City, Hunan Province / 中国血吸虫病防治杂志

Objective To investigate the genetic variation of Eurytrema pancreaticum isolated from goats in Huaihua City, Hunan Province. Methods The partial sequence of mitochondrial cytochrome I (pcox1) and ribosomal 18S rRNA genes were amplified using a PCR assay in E. pancreaticum isolates from goats in Huaihua City, Hunan Province, and the PCR amplification products were sequenced. Then, the gene sequences were subjected to genetic variation and phylogenetic analyses. Results The sequences of the pcox1 and 18S rRNA genes were 430 bp and 1 857 bp in length in 18 E. pancreaticum isolates from goats in Huaihua City, Hunan Province, and there were 14 and 35 variation sites in pcox1 and 18S rRNA gene sequences, with intra-species genetic variations of 0 to 1.4% and 0 to 0.8%, respectively. The sequences of pcox1 and 18S rRNA genes had 99.0% to 99.8% and 99.5% to 99.8% homologies with those from E. pancreaticum Chinese strain recorded in the GenBank database. Consistent phylogenetic analysis results were found based on pcox1 and 18S rRNA genes. The 18 E. pancreaticum isolates from goats in Huaihua City were clustered into a clade with the known E. pancreaticum isolates registered in GenBank, and the clade with these 18 E. pancreaticum isolates was close to the clades with Eurytrema species and far from the clades with other trematodes. Conclusions The E. pancreaticum isolates from goats have a low genetic variation in Huaihua City, Hunan Province. Mitochondrial pcox1 and ribosomal 18S rRNA genes may serve as molecular markers for the studies on the genetic variation in goat-derived E. pancreaticum.

Study on the protective effect and mechanism of adiponectin on hypoxic injury of retinal vascular endothelial cells / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To investigate the protective effect of adiponectin on hypoxia-damaged rhesus monkey choroid /retinal vascular endothelial cells(RF/6A)and related mechanisms. <p>METHODS:<i>In vitro</i> cultured RF/6A cells were randomly divided into the control group, hypoxic injury(induced by CoCl2 stimulation)group and hypoxic injury + adiponectin(5μmol/L, 50μmol/L and 100μmol/L)group. Cell viability was assessed using the MTT assay and optimal concentration of adiponectin was selected. Western blot was used to detect the expression of Bax and Bcl-2 in RF/6A cells. Reactive oxygen species(ROS)detection kit was used to detect the content of ROS in RF/6A cells. <p>RESULTS: Compared with the control group, the cell viability of RF/6A cells in the hypoxic injury group and each adiponectin pretreatment group decreased(all <i>P</i><0.01). Compared with the hypoxic injury group, the cell viability of RF/6A cells in each adiponectin pretreatment group was significantly increased(all <i>P</i><0.05), and adiponectin of 50μmol/L was the appropriate protective concentration. Compared with the control group, the viability of RF/6A cells decreased, the protein expression level of Bax increased, the protein expression level of Bcl-2 decreased, and the content of ROS increased in the hypoxic injury group(all <i>P</i><0.01). Compared with the hypoxic injury group, the viability RF/6A cells increased, the expression level of Bax decreased, the expression level of Bcl-2 increased, and the content of ROS decreased in the adiponectin pretreatment group(all <i>P</i><0.01).<p>CONCLUSION: Our findings suggest that adiponectin can significantly alleviate retinal vascular endothelial cell damage and apoptosis caused by hypoxia, and the mechanism may be related to the inhibition of oxidative stress by adiponectin.

Association of BDNF gene polymorphism with endophenotypes in posttraumatic stress disorder

SUMMARY OBJECTIVE To explore the association of brain-derived neurotrophic factor gene (BDNF) polymorphism with the latent cognitive endophenotype of posttraumatic stress disorder (PTSD) after major natural disasters in Hainan Province, China. METHODS A total of 300 patients with PTSD and 150 healthy controls (HC) were surveyed by psychoanalysis scale to assess their cognitive functions. Polymerase chain reaction (PCR) and polyacrylamide gel electrophoresis (PAGE) were used to detect the BDNF gene polymorphism. RESULTS In terms of the cognitive function, the scores in the PTSD group were worse than those of the HC group (P < 0.05 or P < 0.01). There was a significant difference in the distribution of BDNF genotype and allele frequency between the two groups (P < 0.05). PTSD endophenotypes were significantly different among the BDNF genotypes in the PTSD group (P ≤ 0.01). CONCLUSION There is a statistically significant difference in the polymorphism of BDNF gene between PTSD and HC groups, and the alleles are associated with the incidence of PTSD. Thus, it may be a risk factor for PTSD.

RESUMO OBJETIVO Explorar a associação do polimorfismo do gene fator neurotrófico derivado do cérebro (BDNF) com o endofenótipo cognitivo latente de transtorno de estresse pós-traumático (TEPT) após grandes desastres naturais na província de Hainan, China. MÉTODOS Um total de 300 doentes com TEPT e 150 controles saudáveis (HC) foi investigado pela escala de psicanálise para avaliar as suas funções cognitivas. A reação em cadeia polimerase (PCR) e a eletroforese em gel de poliacrilamida (Page) foram usadas para detectar o polimorfismo do gene BDNF. RESULTADOS Em termos de função cognitiva, as pontuações no grupo TEPT foram piores do que as do grupo HC (P<0,05 ou P<0,01). Houve uma diferença significativa na distribuição do genótipo de BDNF e frequência do alelo entre os dois grupos (P<0,05). Os endofenótipos de TEPT foram significativamente diferentes entre os genótipos de BDNF do grupo TEPT (P≤0,01). CONCLUSÃO Existe uma diferença estatisticamente significativa no polimorfismo do gene BDNF entre o TEPT e os grupos HC, e os alelos estão associados à incidência do TEPT. Assim, pode ser um fator de risco para TEPT.

Measurement of Scars Using Structured-Light 3D Scanning in Forensic Practice / 法医学杂志

Objective To validate the accuracy and reliability of structured-light three-dimensional （SL-3D） scanning in measuring the length and area of the regular and irregular scars on body surface and discuss its value in forensic practice. Methods The lengths of 30 cases of simulated linear scars and 50 cases of linear scars after injury were measured using soft ruler, vernier caliper + thin line method, and SL-3D scanning. The areas of 35 cases of simulated patchy scars and 15 cases of patchy scars after injury were measured using length × width, film tracing with coordinate paper method, pixel method, and SL-3D scanning, and then statistically analyzed. Results The differences between the length of the simulated linear scars measured by SL-3D scanning and standard length had no statistical significance. When simulated patchy scars and patchy scars after injury were measured with high surface curvature and large irregular areas, the differences between the results of SL-3D scanning measurement and the standard area had no statistical significance. When the length of 50 cases of linear scars after injury were measured using SL-3D scanning, the correlation coefficient between the measurement results of two different investigators was 0.998, and the correlation coefficient between the two measurement results by the same investigator was 1.000. The correlation coefficient between the results of SL-3D scanning and that of vernier caliper + thin line method was 0.996. Conclusion The three-dimensional information of the scars on the body surface can be acquired using SL-3D scanning. The measurement of the length and area of the scars is not influenced by the location of scars, curvature of surface, and human factors. The measurement results are accurate, reliable and has unique advantages.

The comparison of robot-assisted laparoscopic and open pyelolithotomy in the management of nonstaghorn renal calculi larger than 2 cm / 复旦学报（医学版）

Objective To investigate the feasibility and safety of robot-assisted laparoscopic pyelolithotomy (RALPL) as the surgical management for nonstaghorn renal calculi larger than 2 cm. Methods Among the patients admitted to our hospital for nonstaghorn renal calculi larger than 2 cm between Jun. ,2013 and Jun. ,2017, a retrospective analysis was performed on the treatment outcome of patients undergoing RALPL (48 cases) and open pyelolithotomy (OPL) (44 cases). The initial stone-free rate, mean operating time, hemoglobin drop, off-bed activity time and hospital stay were compared between two groups. Results Compared with OPL group, RALPL group had a higher initial stone-free rate (91. 66% vs. 70.45%, P < 0.05), and had significant lesser hemoglobin drop, shorter operating time, off-bed activity time and hospital stay. There was no ClavienⅡ or above complications in RALPL group, but there were 2 cases of ClavienⅡcomplications in OPL group. Conclusions Transperitoneal approach RALPL has a better visual angle to observe renal pelvis and calyces, and can ensure a higher initial stone-free rate under the conditions of low intrarenal pelvic pressure and leaving renal parenchyma and vessels intact. RALPL is a minimally invasive, effective and safe treatment, and is an alternative indication for renal calculi larger than 2 cm (including renal pelvis stones with accompanying renal calyceal stones).

Study on HPLC characteristic spectra of Lygodii Herba and chemical compositions of different parts from Lygodii Herba / 中草药

Objective: To establish the HPLC characteristic spectrum of Lygodii Herba from different habitats in Guangxi Province, and to evaluate the difference of components in different parts of medicinal materials. Methods: HPLC was performed on Waters symmetry C18 (250 mm × 4.6 mm, 5 μm) column, with the mobile phase of acetonitrile-0.2% phosphoric acid at flow rate of 1.0 mL/min; Detection wavelength was 354 nm; The column temperature was 30 ℃ and the sample size was 20 μL. Twelve batches of Lygodii Herba samples were determined and the characteristic spectrum of those were established, and the content of rutin, isoquercetin, and astragalin were determined to evaluate the difference of chemical components in different parts of medicinal materials. Results: There were seven characteristic peaks identified in the characteristic spectra of Lygodii Herba samples. Peak 1 was caffeic acid, peak 2 was Rutin, peak 3 was Isoquercitrin and peak 5 was astragalin. The similarities of 11 batches of samples were proved to be higher than 0.900 and one batch of them was proved to be less than 0.900. The chemical component of stem was richer than that of leaves of Lygodii Herba, and the content of the component was higher in the stem than that of the leaves. Conclusion: The method is simple, accurate, and reproducible, which can provide the scientific evidence for controlling the internal quality standards effectively. The preferred harvest season of Lygodii Herba is autumn and winter.

Therapeutic observation of tuina manipulation for lumbar intervertebral disc herniation / 针灸推拿医学（英文版）

Objective:To observe the clinical efficacy of balance-impact tuina therapy for lumbar intervertebral disc herniation (LIDH).Methods:A total of 118 eligible LIDH patients were randomized into an observation group and a control group by the random number table method,with 59 cases in each group.The observation group was intervened by balance-impact tuina therapy,while the control group was intervened by conventional tuina therapy,both for consecutive two weeks.The scores of visual analog scale (VAS),Oswestry disability index (ODI),quality of life questionnaire-core 30 (QOL-C30) were observed before and after treatment;the relapse rate was estimated at the sixth month and twelfth month following the treatment.The data were statistically analyzed.Results:After intervention,the total effective rate was 96.6％ in the observation group versus 91.5％ in the control group,and the between-group difference was statistically significant (P＜0.05).The VAS and ODI scores declined significantly after treatment in both groups (all P＜0.05),and the observation group was markedly lower than the control group (P＜0.05,P＜0.01).The QOL-C30 score increased significantly after treatment in both groups (both P＜0.05),and the observation group was markedly higher than the control group (P＜0.05).The relapse rates at the post-treatment sixth month and twelfth month in the observation group were lower than those in the control group (P＜0.05,P＜0.01).Conclusion:Compared with the conventional tuina therapy,the balance-impact tuina therapy shows advantage in lessening pain,improving the function and enhancing the quality of life in the treatment of LIDH,and it has a lower relapse rate.Thus,this therapy is worth promoting in clinic.

Effect of lipocalin-2 on angiogenesis of murine retinal vascular endothelial cells / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To explore the role of lipocalin-2(LCN-2)in retinal angiogenesis <i>in vitro</i> by observing its effects on proliferation, migration and capillary-like tube formation of murine retinal vascular endothelial cells(RVECs). <p>METHODS: Well cultured RVECs were divided to different groups which were treated with 0, 5, 10 μmol/L LCN-2 for 48h, respectively. Cell proliferation, migration and tube formation were detected by using the EDU assay, transwell assay and matrigel assay, respectively. <p>RESULTS: The cell proliferation rate was promoted in both low and high dose of LCN-2 groups compared to the control cells(<i>P</i><0.05). The number of migrated cells in both LCN-2 groups was significantly larger than that of the control group(<i>P</i><0.05). The number of capillary-like tube structures of both LCN-2 groups was significantly larger than that of the control cells(<i>P</i><0.05). In addition, cell proliferation, migration and tube formation were all increased with the increase of LCN-2 concentration. <p>CONCLUSION: LCN-2 could obviously promote the angiogenesis capacity of RVECs, suggesting that LCN-2 is an important pro-angiogenic factor in retinal angiogenesis.

Analysis of Heterogeneous Biological Characteristics and Prognosis-Related Factors in AL Patients with Mistranslation-Expressed Lymphoid and Myeloid-Related Antigen / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To analyze the heterogeneous biological characteristics of acute leukemia (AL) patients with mistranslation expressed lymphoid and myeloid-related antigens, and it's prognosis-related factors.</p><p><b>METHODS</b>Two hundred and fourteen AL patiens with mistranslation expressed lymphoid and myeloid-related antigens were grouped according to immunophenotypes, and the heterogeneous biologic charecteristics and prognosis related factors were analyzed, moreover the survival curves were drawn to analyze the survival of patiens.</p><p><b>RESULTS</b>The immunophenotype in 214 cases was mainly cross-expression of myeloid and B lineage antigen (118 cases), followed by cross-expression of myeloid antigen and T lineage (88 cases), while the cross-expression of myeloid, T and B lineages, was less (only 8 cases). In ALL patiens with cross-expression of myeloid antigen, the CD33 was main type; while in AML patients with cross-expression of lymphoid antigen, CD7 was main type of lineage antigen, CD19 was main type of B lineage antigen. Among 214 AL patients, the cross-expression of CD55 and myeloid antigen was found in 30 cases, the cross-expression of CD7, CD19 and CD74 was observed in 6 cases, the cross-expressions of CD7, CD34 and CD56 was detected in 4 cases. Among AML patients with lymphoid antigen expression, the recurrent chromosmal abnormalities were found in 16 cases; among ALL patients with myeloid antigen expression, the recurrent chromosomal abnormalities were observed in 10 cases. The mistranslation antigen expression existed in 26 patients with recurrent chromosomal abnormalities, the mistranslated CD33 and CD13 in ALL patients with myeloid antigen expression was common, while the mistranslated CD2, CD56 and CD19 in AML patients with lymphoid antigen expression was common. As compared with patients without lymphoid antigen expression, the survival rate decreased significantly in patients with mistranslated CD7(+) and CD34(+) (both P<0.05). The logistic regression analysis showed that CD7, CD34 were main influencing factors for prognosis of AL patients (both P<0.05).</p><p><b>CONCLUSION</b>The AL with mistranslation expressed lymphoid and myeloid antigens is a special kind of leukemia which possesses the heterogencous biological characteristcs and unique prognostic features, thus the immunophemotype of AL patients should be detected by flow cytometry. The existance of mistranlation-expressed differatiation antigens such as CD7 and CD34 is mainly influencing factors for the prognosis of AL patiens.</p>

Implant prosthesis design scheme for sophisticated congenital partial edentia / 华西口腔医学杂志

Sophisticated congenital partial edentia are often accompanied by severe shortage of bone height and width due to the absence of permanent teeth; such condition will affect implant placement. This study aimed to display the different typical implant rehabilitation schemes we designed for sophisticated congenital partial edentia cases with severely atrophic alveolar bone.

RNA-binding protein HuR regulates hsa-let-7c expression by its RNA recognition motif / 生理学报

MicroRNAs (miRNAs) are small noncoding RNAs that control diverse cellular and developmental events through repression of large sets of target mRNAs. miRNAs expressions were mainly regulated at two levels: transcriptional and post-transcriptional. Transcriptional regulation of miRNA-encoding genes produce specific expression patterns of individual miRNA. However, the mechanism of post-transcriptional regulation of miRNAs remains largely unknown. The present study was aimed to clarify whether HuR, an evolutionary conserved AU-rich binding protein, could regulate miRNAs expressions. By means of a computational screen for AUUUA motifs within pri-miRNAs, we found that the downstream of hsa-let-7c but not hsa-miR-21 was enriched of AUUUA motifs. Then we transfected HuR and mutant HuR lacking RNA recognition motif 3 (RRM3) respectively into HEK293T cells. And HuR protein and miRNAs expressions were detected by Western blot and real-time PCR, respectively. The results showed that the overexpression of HuR promoted mature hsa-let-7c expression but not hsa-miR-21 expression. Furthermore, overexpression of HuR deletion mutant lacking RRM3 did not promote hsa-let-7c expression. These results suggest that RRM3 is crucial for HuR mediating mature hsa-let-7c expression. Collectively, these findings proposed a novel role of HuR in biogenesis of miRNAs, possibly by way of post-transcriptional regulation of miRNAs.

Influence of body mass index on postoperative complications and survival in esopha-geal squamous cell carcinoma patients based on propensity score matching method / 中国肿瘤临床

Objective: To explore the influence of body mass index (BMI) on postoperative complications and survival in patients with esophageal squamous cell carcinoma (ESCC) using propensity score matched (PSM) methods. Methods: We retrospectively analyzed clinical data of 533 patients with ESCC who were admitted for thoracic surgery in Shandong Provincial Hospital, between January 2011 and December 2012. After conducting PSM methods to balance the covariates, the incidence of postoperative complications and sur-vival rate were compared between the two groups. The effects of BMI on postoperative complications and survival were analyzed with multivariate Logistic regression and Cox proportional hazard analyses, respectively. Survival analysis was performed using Kaplan-Mei-er curves and Log-rank test. Results: After adjusting the PSM and confounding variables, the two groups were well matched (146 pa-tients each) without significant differences in baseline characteristics. The incidence of wound infection and respiratory system compli-cations in the H-BMI group (BMI≥25 kg/m2) were significantly higher than those in the N-BMI group (18.5-25 kg/m2) (P<0.05). H-BMI was an independent risk factor for wound infection and respiratory system complications. In the present study, no significant differ-ence was observed in the 3-year overall survival between H-BMI and N-BMI patients (54.1% vs. 47.8%, P=0.212). Patients with H-BMI had significantly better 5-year overall survival than those with N-BMI (39% vs. 25%, P=0.016). The subgroup analysis showed that pa-tients with H-BMI had a better overall survival than those with N-BMI in stages Ⅰ-Ⅱ (47.3% vs. 29.0%, P=0.032). However, this difference was not significant when patients were stratified into stages Ⅲ and Ⅳ (24.5% vs. 16.7%, P=0.393). Conclusions: H-BMI appears not to decrease the overall survival of patients with ESCC. Therefore, ESCC in patients with H-BMI can receive surgical treatment safely, but proper intraoperative management and close postoperative monitoring should be performed.

Combination of musk and olibanum promotes proliferation of prostate stem cells in mice: An experimental study / 中华男科学杂志

Objective@#To study the effects of muskolibanum combination on the proliferation and differentiation of prostate stem cells.@*METHODS@#We cultured prostate epithelial cells and urogenital sinus mesenchymal (UGSM) cells from 7－10 d old C57BL/6 mice and 16－18 d old pregnant C57BL/6 mice, transplanted the mixed suspension of the two types of cells under the kidney envelope of SCIDCB.17 male mice, and harvested the transplants 30 days later. We randomly divided the SCIDCB.17 mice into four groups to be treated intragastrically with musk (n = 8), olibanum (n = 8), musk+olibanum (n = 7), and normal saline (blank control, n = 8)) respectively, all for 14 days. Then we collected the kidney tissue for observation of the morphology of the glandular tubes and differentiation of different subsets of stem cells by HE staining and determination of the expressions and distribution of P63, CD133, CD117 and Sca1 by immunohistochemistry and Western blot.@*RESULTS@#A system was successfully established for the isolation and mixed culture of Sca1 Lin+ CD49f+ (LSC) cells of prostate stem cells and UGSM cells of the mouse embryonic prostate. Immunohistochemistry showed positive expressions of P63, CD133, Sca1, and CD117 in the prostatic acinar epithelia and proved the presence of prostatic acinar epithelial structure in the transplants. Compared with the blank control group, the expressions of CD133, Sca1 and CD117 were significantly increased in the musk, olibanum, and musk+olibanum groups (P< 0.05), higher in the musk+olibanum than in the musk or olibanum group (P< 0.05), and their protein expressions were even more elevated in the musk+olibanum group (P< 0.01), with statistically significant difference from the olibanum group (P< 0.05).@*CONCLUSIONS@#The combination of musk and olibanum can improve the proliferation and differentiation of prostate stem cells.

Retrospective analysis of operative methods and complications in 29 patients with carotid body tumor / 临床耳鼻咽喉头颈外科杂志

Objective:To analyze the operative methods and complications for surgical resection of carotid body tumor (CBT). Method:Clinical data of 29 patients underwent CBT resection from Jan 2007 to Apr 2015 were retrospectively studied. Result:Five out of 29 patients got bilateral CBT, the others had unilateral lesions, totally 34 CBT resection were performed. Tumor completely dissected without carotid artery clamping and reconstruction in 18 procedures, tumor resection combined with external carotid artery resection in 16 procedures, 9 out of these 16 procedures combined with internal carotid artery resection. The internal carotid artery was reconstructed with autologous greater saphenous vein in 4 procedures, with artificial graft in 3 procedures, and internal carotid artery ligation without reconstruction in 2 procedures. There was no patient death during perioperative period, cerebral infarction happened in 1 patient and cranial nerve injury occurred in 12 cases. Conclusion:Carotid artery resection and reconstruction is very common during CBT resection, surgeon must be familiar with the methods of carotid artery reconstruction, and the most common complication of carotid body tumor resection is the cranial nerve injury.