Application of Self-assembled Nano-strategies of Traditional Chinese Medicine in Tumor Therapy： A Review / 中国实验方剂学杂志

Chinese medicine self-assembly nano-strategies（CSAN） is to utilize the self-assembly property of Chinese medicine components， so that the Chinese medicine components can self-assemble to form structurally stable nano-preparations through non-covalent interactions. The formation of Chinese medicine self-assembly nano-preparations is often a synergistic result of a variety of non-covalent interactions， and many Chinese medicine monomers are susceptible to self-assembly due to their structural characteristics， and the phenomenon of self-assembly of Chinese medicine is also common in the decoction of single or compound Chinese medicine， which has attracted the attention of researchers. It is found that CSAN can improve the solubility and bioavailability of active components in Chinese medicine， which is of positive significance for the development and application of insoluble components of Chinese medicine. The self-assembly phenomenon of Chinese medicine decoction is closely related to the therapeutic efficacy， and the study of self-assembly phenomenon of Chinese medicine will bring a new perspective for the explanation of the mechanism of Chinese medicine decoction. At the same time， traditional Chinese medicine（TCM） has unique advantages in the field of anti-tumor. The application of CSAN in the field of oncology can not only exert the anti-tumor effect of the active components of Chinese medicine directly， but also act as a natural nano-carrier to carry chemotherapy drugs for combination chemotherapy， improve the targeting of drugs， enhance the anti-tumor efficacy， and reduce the side effects of chemotherapy， which has excellent anti-tumor potential. The preparation method of Chinese medicine self-assembly nano-preparations is simple， low cost， and has better safety than traditional nano-preparations， which is conducive to the promotion of the clinical transformation of nano-preparations， and also helps to provide new strategies and perspectives for promoting the modernization of TCM. Therefore， based on a large number of researches in this field in recent years， this paper reviewed the formation mechanism， different assembly forms， formation conditions and stability of Chinese medicine self-assembly nano-preparations by searching databases such as China national knowledge infrastructure（CNKI）， PubMed， WanFang data and VIP， and summarized the application of CSAN in different tumor therapies， providing a reference for further research on CSAN.

Clinical characteristics and genetic analysis of a pedigree with Stargardt disease caused by a novel mutation in ABCA4 gene / 中华检验医学杂志

Objective:To explore the clinical characteristics and genetics of a pedigree with Stargardt disease, and investigate the pathogenicity of ABCA4 (ATP binding cassette subfamily A member 4) gene mutations in Stargardt disease.Methods:The proband was admitted to the Second People′s Hospital of Jinan in May 2021 due to diminution of vision. The proband was diagnosed with Stargardt disease according to the clinical diagnostic criteria of Stargardt disease. Detailed ophthalmological examinations was also performed on family members of the proband. Genomic DNA were extracted from the proband and the family members, and the whole exon sequencing was performed to find pathogenic gene mutations. The hazard of mutations was analyzed by polyphen-2, SIFT and MutationTaster websites. Sanger sequencing was used to verify the mutations. Conserved analysis of homologous species and 3-dimensional (3D) molecular model of the protein were used to analyze the pathogenicity.Results:Ophthalmological examinations showed reduced binocular vision, macular atrophy and "bull′s eye sign" in the proband and there was no abnormal signs and symptoms among the family members. Through whole exon sequencing analysis and Sanger sequencing verification, the compound heterozygous mutations (c.215G>A and c.6563T>C) of ABCA4 gene were co-segregated with this disease in this family. SIFT, Polyphen-2 and MutationTaster predicted that these two mutations were pathogenic. Conservative analysis and 3D molecular model of protein showed that mutations could cause changes in protein structure and affect protein function.Conclusion:The compound heterozygous mutations (C.215G>A and C.6563T>C) of ABCA4 gene are the pathogenic mutations of Stargardt disease in this pedigree.

Analysis of occupational health examination of workers in a nuclear power plant in Guangdong, China / 中国辐射卫生

@#<b>Objective</b> To analyze the abnormality of radiation damage sensitive indices in radiation workers after operation of a third-generation nuclear power plant in Guangdong Province, China, and to provide a basis for promoting the occupational health management of radiation workers. <b>Methods</b> A two-way cohort study (2019—2021) was conducted to compare the radiation sensitive indices of occupational health examination in the exposed group (453 subjects) and the control group (160 subjects). <b>Results</b> In 2021, the free triiodothyronine (FT3) level of the exposed group was significantly higher than that of the control group [(5.57 ± 0.56) pmol/L <i>vs</i> (5.42 ± 0.60) pmol/L, <i> t</i> = 0.59, <i>P</i> < 0.05]. From 2019 to 2021, the exposed group showed significant changes in the average levels of platelet, hemoglobin, FT3, free thyroxine (FT4), and thyroid-stimulating hormone (<i>P</i> < 0.05); FT3 and FT4 first increased and then decreased, while TSH decreased continuously. <b>Conclusion</b> Long-term exposure to low-dose ionizing radiation from nuclear power plants can affect the platelet, hemoglobin, FT3, FT4, and thyroid-stimulating hormone of radiation workers, and the effect is relatively prominent on thyroid function by causing a tendency to hypothyroidism.

Bushen Jiedu Prescription Inhibits Metastasis of Colorectal Cancer by Regulating Polarization of M2-TAMs in Vivo / 中国实验方剂学杂志

ObjectiveTo explore the inhibitory effect of Bushen Jiedu prescription （BSJDP） on the metastasis of colorectal cancer （CRC） via activation of M2 tumor-associated macrophages （M2-TAMs） in vivo. MethodThe model of xenograft tumor was established with C57BL/6 mice， and then the model mice were randomly assigned into blank control group， Bushen Jiedu recipe-low dose （11.2 g·kg-1） group （BSJDP-L）， and Bushen Jiedu Recipe-high dose （22.4 g·kg-1） group （BSJDP-H）. Tumors were abolished when the volume reached 1.5-2.0 cm3. The mice were sacrificed 28 days post tumor abolishing and then the lung metastasis was observed. Histopathological changes in lung metastasis were examined by hematoxylin-eosin （HE） staining of metastasis tissues， and immunofluorescence （IF） staining was employed to observe the effect of BSJDP on tumor apoptosis and hypoxia. Flow cytometry was performed to analyze the macrophages M1/M2 ratio of tumor tissue. The expression levels of the genes （Arg1， CD206， and CD163） associated with the polarization of M2-TAMs were determined by Real-time fluorescent quantitative polymerase chain reaction （Real-time PCR）. ResultThe metastasis rate was 70%， 40%， and 10% in the blank control group， BSJDP-L group， and BSJDP-H group， respectively. The lower metastasis rates of BSJDP-L and BSJDP-H groups proved that BSJDP significantly inhibited lung metastasis of CRC. Compared with the blank control group， BSJDP-L and BSJDP-H reduced the tumor cell infiltration in tumor tissue （P<0.01）， increased the apoptosis of tumor cells， alleviated the hypoxic environment， and down-regulated the expression of Arg1， CD206， and CD163 in the tumor tissue （P<0.01）. In addition， the ratio of M2 macrophages ranked in a descending order of blank control group （34.867%） > BSJDP-L group （22.033%） > BSJDP-H group （11.633%） （P<0.01）. ConclusionBSJDP inhibits the lung metastasis of CRC by inhibiting the activation of M2-TAMs in the tumor microenvironment.

Advances in study of immune-related small fiber neuropathy / 中华神经科杂志

Small fiber neuropathy is a peripheral neuropathy that selectively involves unmyelinated C-type fibers and (or) thin myelinated Aδ-type fibers. It is mainly manifested as abnormal pain and temperature perception and autonomic symptoms. Changes in certain inflammatory markers and antibodies related to autoimmune diseases can occur in patients with immune-related small fiber neuropathy. This article aims to explore the immunological progress in small fiber neuropathy.

Mental health status among non-psychiatric inpatients in a general hospital / 四川精神卫生

ObjectiveTo analyze the mental health status of non-psychiatric inpatients in a general hospital， and to explore the relevant influencing factors， so as to provide references for the screening of mental health problems and the construction of intervention models among non-psychiatric inpatients. MethodsA cross-sectional study was conducted among 916 non-psychiatric inpatients in a third grade class A general hospital in Guangzhou， and all the inpatients were assessed using Patient Health Questionnaire-9 item （PHQ-9）， Generalized Anxiety Disorder-7 item （GAD-7）， Athens Insomnia Scale （AIS） and Columbia-Suicide Severity Rating Scale （C-SSRS） to detect their depression， anxiety， insomnia and suicide risk status. Thereafter， univariate and multivariate Logistic regression analysis were used to screen the risk factors affecting the mental health of inpatients. ResultsA total of 339 （37.0%） inpatients with positive mental health problems were screened， and the screening results for each dimension revealed 218 cases （23.8%） of depression， 141 cases （15.4%） of anxiety， 257 cases （28.1%） of insomnia， 42 cases （4.6%） of suicidal ideation and 7 cases （0.8%） of suicidal behavior. Binary Logistic regression analysis showed that female （OR=1.379， P<0.05） was a risk factor for positive screening of mental health problems. Ordinal Logistic regression analysis denoted that age above 60 years old （OR=1.542， P<0.05） and singlehood （OR=2.055， P<0.05） were risk factors affecting the severity of depression， while senior high school to junior college education （OR=0.524， P<0.05） was a protective factor of depression， meantime， female （OR=1.472， P<0.05） was a risk factor affecting the severity of insomnia. ConclusionMental health problems are quite common among non-psychiatric inpatients in general hospitals， and are mainly affected by factors such as gender， age， marital status and educational background.

Differential miRNA expression in the peripheral blood of patients with Keshan disease and its mechanism / 中华地方病学杂志

Objective:Through differential miRNA expression profiles and bioinformatics in the peripheral blood of patients with Keshan disease (KD) and healthy control, to explore the possible pathogenesis of KD.Methods:Ten patients with chronic KD (KD group) were selected in the severe disease area of KD in Wulian County, and 10 healthy subjects (control group) were selected in non-KD area of Dongchangfu District, Shandong Province. Blood sample of elbow vein was collected and plasma was separated. RNA-seq technology was used to construct the differential expression profiles of miRNA in KD and control groups. Target mRNAs were screened using Starbase, miRTarBase, miRDB and TargetScan. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were conducted to investigate the possible pathogenesis of KD.Results:Compared the control group and KD group, 132 differentially expressed miRNAs were screened out, including 90 upregulated and 42 downregulated miRNAs. Through Starbase, miRTarBase, miRDB and TargetScan, 53 miRNAs were obtained, 737 targeted mRNAs were obtained. GO analysis showed that the differential genes were mainly involved in the biological processes of Ras protein signal transduction, transmembrane transport, cell cycle regulation, cell adhesion, etc. KEGG pathway analysis showed that the differential genes were mainly involved in viral infection, endocytosis, adhesion spot and actin regulation.Conclusion:In this study, RNA-seq technology is used to obtain differential miRNA expression profiles of KD patients and healthy control, and target pathogenic genes and signaling pathways that may be related to KD are screened out.

Evaluation of a new ultrafast real-time fluorescence polymerase chain reaction and common assays in the detection of novel Bunya virus / 中华传染病杂志

Objective:To investigate the specificity and sensitivity of four methods including ultrafastreal-time fluorescence polymerase chain reaction (PCR), real-time fluorescence (RT)-PCR, enzyme-linked immunosorbent assay (ELISA) and gold immunochromatography assay (GICA) for the detection of novel Bunya virus, so as to provide experimental basis for the early diagnosis of severe fever with thrombocytopenia syndrome (SFTS).Methods:Serum samples from 86 clinically diagnosis SFTS patients admitted to the Jinan Infectious Diseases Hospital Affiliated to Shandoug University were tested by ultrafast real-time fluorescence PCR, RT-PCR, ELISA and GICA during June 1 to September 30, 2017. Chi-square test was used for statistical analysis.Results:Among 86 serum samples, the positive rate of novel Bunya virus of ultrafast real-time fluorescence PCR, RT-PCR, IgM-ELISA, IgG-ELISA, IgM-GICA and IgG-GICA were 82(95.34%), 79(91.86%), 41(47.67%), 8(9.3%), 19(22.09%) and 3(3.49%), respectively. The specificity of ultrafast real-time fluorescence PCR was 100%, and the sensitivity was 1×10 3 copies/mL.Repeated amplification test showed that the variation coefficient of the computed tomography value was <2%.During phases one, two and three, the positive rates of ultrafast real-time fluorescence PCR were 41(97.62%), 34(94.44%) and 7(87.50%), and RT-PCR were 39(92.86%), 33(91.67%) and 7(87.50%), respectively. During phases one and two, the positive rate of ultrafast real-time fluorescence PCR was slightly higher. The positive rate of anti-novel Bunya virus antibody (IgM) tested by ELISA had a significant increase from phase one (28.57%)to phase three (87.50%). There were statistical differences between phase two and phase, as well as between phase three and phase one ( χ2=8.347 and 7.561, respectively, both P<0.01). IgM-GICA also had an increase from phase one (14.29%) to phase two (33.33%)( χ2=3.962, P<0.01), while it was still lower than the other tests.In phase one, the positive rate of RT-PCR was higher than those of ELISA(both IgM and IgG)and GICA(both IgM and IgG)( χ2=33.740, 55.080, 49.010 and 64.340, respectively, all P<0.01). In phase two, the positive rate of RT-PCR was higher than those of ELISA(both IgM and IgG)and GICA(both IgM and IgG) ( χ2=7.700, 46.720, 23.700 and 50.630, respectively, all P<0.01). In phase three, the positive rates of ultrafast real-time fluorescence PCR, RT-PCR and IgM-ELISA were equivalent, which were all higher than those of IgG-ELISA and GICA (both IgM and IgG). The positive rates of RT-PCR and IgG-ELISA, IgM-GICA and IgG-GICA were significantly different (all χ2=6.250, all P<0.05). Conclusion:In the early detection of novel Bunya virus, ultrafast real-time fluorescence PCR has higher sensitivity, specificity, good repeatability and high stability, which greatly reduces the amplification time compared with the traditional RT-PCR, and is of great value in the early and rapid diagnosis of SFTS.

Chloroxoquinoline inhibits invasion in breast cancer via down-regulating Rho/Rho kinase signaling pathway / 浙江大学学报·医学版

OBJECTIVE: To investigate the effect of chloroxoquinoline on cytoskeleton of breast cancer cells and its relation with Rho/Rho kinase signaling pathway. METHODS: Breast cancer Bcap37 and MDA-MB-453 cells were treated with different concentrations of chloroxoquinoline. Wound healing and Transwell assay were conducted to detect cell migration and invasion, respectively. Rhodamine-phalloidin staining and immunofluorescent staining were used to observe the polymerization state of F-actin and the expression of α-Tublin in breast cancer cells, respectively. Western blot was used to detect the phosphorylation level of key protein in Rho/Rho kinase signaling pathway. RESULTS Compared with the control group, chloroxoquinoline treatment induced dose-dependent decrease in cell migration and invasion, and Bcap37 and MDA-MB-453 cells treated with chloroxoquinoline showed dose-dependent changes in cell morphology and decrease in cell body. The staining of F-actin and α-Tublin was irregular and clustered. Furthermore, treatment of chloroxoquinoline down-regulated the phosphorylation of the Rho/Rho kinase signaling proteins Cofilin, Limk and Rock2 (all P<0.01). CONCLUSIONS Chloroxoquinoline inhibits the cytoskeleton in breast cancer Bcap37 and MDA-MB-453 cells and inhibits cell migration. This effect may be associated with down-regulation of Rho/Rho kinase signaling pathway.

A molecular epidemiological survey on the outbreak of recombinant Norovirus GII.P16-GII.2 in Guangzhou city / 中华实验和临床病毒学杂志

Objective@#To find out the source and the epidemic pattern of norovirus outbreak in July, 2016 to June, 2017 in Guangzhou.@*Methods@#The stool samples and clinical information of diarrhea cases were collected by the sentinel hospitals and CDCs; a real-time RT-PCR method was used to detect the norovirus nucleic acids from the samples, the positive ones were amplified and sequenced; the partial sequences of norovirus were aligned by an online BLAST alignment, and a phylogenetic tree was constructed by a neighbor-joining method .@*Results@#A total of 854 cases with infectious diarrhea were reported by Guangzhou diarrhea surveillance network from July, 2016 to June, 2017; the gender ratio (male versus female) was 1∶0.67; 78.33% of the cases were preschool children under the age of 7 years. Totally 220 samples were detected norovirus G II+ (25.76%, including 5 double-positive samples with G I+ ). GII.Pe-GII.4.Sydney_2012 was the prevalent genotype in the second half of 2016 (94.64%), which was replaced by GII.P16-GII.2 in the first half of 2017 (67.65%). Since September 2016, the reported number of norovirus-caused diarrhea epidemic was increased gradually; the peak of epidemic curve emerged in February to March of 2017, and the number started to decrease since April. In May to June there were only 2-3 epidemics reported monthly. All the endemics from September to November 2016 were caused by genotype GII.Pe-GII.4.Sydney_2012; the endemics from December 2016 to April 2017 were mainly caused by genotype GII.P16-GII.2. Some samples from kitchen workers and babysitters were detected GII+ , which was consistent with the result of the cases′ samples.@*Conclusions@#It was the first time that the novel GII.P16-GII.2 recombinant strain outbroke occurred in Guangzhou City and homology analysis also suggested that GII.P16-GII.2 was the main source of those epidemics in 2016 -2017 winter and spring season. Furthermore, The kitchen workers and babysitters may have played an important role in the spread of norovirus.

Comparative analysis of lncRNA-mRNA co-expression between Keshan disease and dilated cardiomyopathy / 中华地方病学杂志

Objective By constructing the differential expression profile of lncRNA/mRNA in peripheral blood plasma of patients with Keshan disease (KSD) and dilated cardiomyopathy (DCM),to explore the commonality and characteristics of the two diseases in molecular mechanism.Methods Ten patients with chronic KSD were selected in the severe disease area of KSD in Shandong Province,and 10 cases of DCM and 10 healthy subjects (control group) were selected in non-KSD area.Blood of elbow vein was collected and plasma was separated.RNA-seq technology was used to construct the differential lncRNA/mRNA expression profile between KSD and control group,DCM and control group,and co-expression and specific expression of partial genes in KSD and DCM were analyzed through Wien analysis.The lncRNA-mRNA co-expression network maps of specific part of KSD,specific part of DCM and common part of the two diseases were constructed,and Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were applied to distinguish the biological function of the two diseases.Results Compared with control group,102 dysregulated mRNAs and 22 dysregulated lncRNAs showed the same trend in KSD and DCM.And 3 606 mRNAs and 451 lncRNAs were only differentially expressed in KSD group,217 mRNAs and 137 lncRNAs were only differentially expressed in DCM group.The differentially expressed lncRNA/mRNA shared between the KSD and DCM groups were mainly about viral transcription,immuno-inflammatory response,oxidative stress signaling pathways.The KSD specific lncRNA/mRNA mainly participated in cell membrane damage and viral myocarditis.The DCM specific lncRNA/mRNA mainly regulated mitochondrial structure and oxidative phosphorylation related enzymes.Conclusion The differentially expressed lncRNA/mRNA shared in KSD and DCM groups are mainly involved in viral transcription,oxidative stress signaling pathways;KSD specific lncRNA/mRNA are mainly related to cell membrane damage and viral myocarditis;DCM specific lncRNA/mRNA mainly regulate mitochondrial structure.

Preparation of antibody against ANKRD22 and its expression in colorectal cancer / 预防医学

Objective@# To prepare the monoclonal antibody against ankyrin repeat domain 22（ANKRD22）and to investigate its expression in colorectal cancer tissues.@*Methods @#The recombinant human ANKRD22 was expressed through E. coli and pET-42a and then used to immunize Balb/c mice after purification. Anti-human ANKRD22 specific monoclonal antibodies were selected by Western blotting with 293T cell lysate highly expressing ANKRD22 as antigen. The expression of ANKRD22 in the tissue microarrays of 112 patients with colorectal cancer was detected by immunohistochemical staining.@*Results @#Four specific monoclonal antibodies against human ANKRD22 were screened out of 93 hybridoma cells,which reacted well with natural human ANKRD22. ANKRD22 was mainly distributed in the cytoplasm of colorectal cancer cells. In 112 cases of colorectal cancer,94 cases were detected positive for ANKRD22 expression,with the positive rate of 83.93%. The expression of ANKRD22 was statistically correlated with the expression of p53 and β-catenin（P<0.05）,but not with age,sex,location of tumors,AJCC stage,Dukes stage,degree of differentiation,lymph node metastasis and mismatch repair gene expression（P>0.05）.@*Conclusion @#The expression level of ANKRD22 was high in colorectal cancer. ANKRD22 might be involved in the carcinogenesis of colorectal epithelium and be a potential diagnostic marker.

Treatment of vertical femoral neck fractures by the technique of static compression screws with medial support / 中华创伤骨科杂志

Objective To investigate the advantage of the technique of static compression screws with medial support using one ordinary cannulated compression screw (OCCS) and 2 headless cannulated compression screws (HCCSs) in reducing complications in the treatment of vertical femoral neck fractures.Methods From December 2014 to July 2017,79 patients were treated at Department of Orthopaedics,The Sixth People's Hospital of Shanghai for vertical femoral neck fractures.They were 51 men and 28 women,aged from 20 to 65 years (average,49.1 years).Their injury involved 45 left sides and 34 right sides.Of them,37 were treated with one OCCS at the top and 2 HCCSs at the bottom of a triangle arrangement for fixation of the vertical femoral neck fracture (the experimental group);the other 42 were treated with 3OCCSs at a triangle arrangement for fixation of the vertical femoral neck fracture (the control group).Their fracture healing and complications were followed up at postoperative 6 weeks,3,6 12,18,24 months and any time of discomfort by anteroposterior and lateral X-ray films of the knee joint.Results The 2 groups were compatible due to insignificant differences between their preoperative general data (P ＞ 0.05).This cohort was followed up for 9 to 24 months (average,17.5 months).Of them,52 achieved fracture union.Of the 27 patients who failed,8 were in the experimental group (21.6％) and 19 in the control group (45.2％),showing a significant difference in the rate of failure between the 2 groups (P ＜ 0.05).In the experimental group,the rate of nonunion was 8.1％ (3/37),the rate of implant failure 18.9％ (7/37),and the rate of fermoral neck varus 8.1％ (3/37),all significantly lower than those in the control group [26.2％ (11/42),40.5％ (17/42) and 23.8％ (13/42),respectively] (P ＜0.05).Conclusion For treatment of vertical femoral neck fractures,the technique of static compression screws with medial support is not only easy but also leads to a lower rate of complications.

Construction of a lentivirus vector containing Pax6 and its transfection of human bone marrow mesenchymal stem cells / 中国组织工程研究

BACKGROUND:Pax6 gene plays an important role in eye development and differentiation, and to study how it regulates the differentiation of human bone marrow mesenchymal stem cel s (BMSCs), gaining the BMSCs stably over-expressing Pax6 is crucial, which is also the basis of stem cel replacement therapy. OBJECTIVE:To construct a lentivirus vector containing Pax6 and detect the expression of Pax6 in transfected human BMSCs. METHODS:Pax6 gene was extracted using PCR. After its connection with lentivirus vector pHIV-EGFP, it was then packaged by 293T cel s. The human BMSCs were transfected with recombinant lentivirus Pax6-EGFP as wel as lentivirus vector pHIV-EGFP, which was considered negative control group. The cel ular morphology was observed by a fluorescence microscope, and the mRNA expression of Pax6 was detected by real-time PCR. RESULTS AND CONCLUSION:The recombinant lentivirus Pax6-EGFP was constructed successful y with a titer of 3×109 pfu/L. After the transfection, both the green fluorescent protein and Pax6 gene were expressed detected using fluorescence microscope and real-time PCR, showing that the method of lentiviral transfection is a safe and effective way to modify BMSCs.

Expression of recombinant phosphodiesterases 3A and 3B using baculovirus expression system

Background: Phosphodiesterase 3A [PDE3A] and phosphodiesterase 3B [PDE3B] play a critical role in the regulation of intracellular level of adenosine 3',5'-cyclic monophosphate [cyclic AMP, cAMP] and guanosine 3',5'-cyclic monophosphate [cyclic GMP, cGMP]. Subsequently PDE3 inhibitors have shown to relax vascular and inhibit platelet aggregation in cardiovascular disease

Objectives: In this study, our aim was to establish a method of expression for the recombinant human PDE3A and PDE3B proteins in insect cells using baculovirus expression system in order to investigate the activity of the expressed PDE3A and PDE3B proteins

Materials and Methods: The full length human PDE3A and PDE3B cDNA were cloned into recombinant baculovirus and transfected into the SF9 insect cells. Recombinant proteins were collected at 48 h, 60 h, 72 h, and 84 h post transfection. Transfection of recombinant baculovirus was verified by the morphological changes of the SF9 cells. Expression of human PDE3A and PDE3B was detected by using RT-PCR and western blot, respectively. The [125]I RIA method was used to determine the level of adenosine 3',5'-cyclic monophosphate cAMP and cGMP, correspondingly. The activity of the expressed PDE3A and PDE3B proteins were investigated by cAMP and cGMP dsgradation with or without addition of milrinone, a potent and selective PDE inhibitor

Results: Recombinant human PDE3A and PDE3B proteins were stably expressed in SF9 cells and could be detected by distinct morphological changes in the SF9 cells, RT-PCR, and western blot at 48 h post-transfection. The molecular weights of the recombinant PDE3A and PDE3B molecular weights proteins were about 120 KDa and 135 KDa, respectively. Results of [125]I RIA assay showed that the levels of cAMP and cGMP were significantly decreased after incubation with the expressed PDE3A and PDE3B proteins. Furthermore, degradation of cAMP and cGMP through the activity of PDE3A and PDE3B was suppressed following to the addition of milrinone

Conclusions: Recombinant human PDE3A and PDE3B could be expressed in SF9 cells using baculovirus expression system, and thus provides the basic material for studying human PDE3A and PDE3B activity

Analysis of the drug resistance and the integron resistance gene cassette's characteristics of Shigella flexneri / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the correlation between Shigella flexneri multi-drug resistance and drug resistance gene cassette of integrons.</p><p><b>METHOD</b>All 79 strains of Shigella flexneri were isolated from the feces of children ranged in age from 6 months to 14 years in some hospitals of Jinan, between May 2009 and April 2012.The resistance was detected by Kirby Bauer agar diffusion method, 1, 2 and 3 integron gene was amplified by PCR, the variable region of positive strains treated with enzyme digestion and determined by Series Analysis.</p><p><b>RESULT</b>Among 79 Shigella flexneri strains, the resistance rate was 91% (72/79) to ampicillin, chloramphenicol, tetracycline, streptomycin, 70% (55/79) to sulfamethoxazole/trimethoprim, 30% (24/79), 23% (18/79), 33% (26/79) and 32% (25/79) to cefotaxime, ceftazidime, ciprofloxacin and levofloxacin.All 79 strains were susceptible to cefoxitin, imipenem, cefoperazone/sulbactam. The common drug resistance pattern is ampicillin tetracycline-chloramphenicol-streptomycin, accounted for 91% (72/79); 91% (72/79) strains carried integrons of class 1, 86% (68/79) strains carried integrons of class 2, No intI3 was detected. The resistance to ampicillin, streptomycin, tetracycline, chloramphenicol of atypical class 1 integron positive strains was significantly higher than the negative strains (χ² = 35.96, P<0.01). The sequencing results:dfrV was detected in class 1 integron variable regions of 9 strains, dfrA17-aadA5 in 2 strains, blaOXA-30-aadA1 in 70 strains, 2 strains were not detected resistance gene cassette, all resistance gene cassettes were dfrA1-sat1-aadA1 in class 2 integron variable regions.</p><p><b>CONCLUSION</b>The muti-drug resistance of Shigella flexneri in Jinan was closely associated with integrons.</p>

Clinical efficacy of Shuxuening injection combined with epalrestat for diabetic nephropathy / 中国生化药物杂志

Objective To explore the clinical efficacy of Shuxuening combined with epalrestat in the treatment of diabetic nephropathy. Methods Eighty patients from March 2014 to June 2015 in Metabolic Disease Department of the Second Hospital of Tianjin were randomly divided into observation group and control group with 40 patients in each group.The control group received epalrestat, and the observation group received Shuxuening injection on the baisis of control group. The related indicators were compared between two groups.Results After treatment, the 24 h-urine microalbumin, plasma low density lipoprotein and serum creatinine in observation group [(239.31 ±106.54)mg/L,(2.45 ±0.55)mmol/L,(95.54 ± 22.13)mol/L]were lower than those in control group [(349.90 ±148.40),(3.41 ±0.52),(108.76 ±34.30)](P<0.05).The β2-microglobulin (β2-MG) and plasma high-sensitive C-reactive protein (hs-CRP) in observation group [(0.39 ±0.06),(6.31 ±1.58)mg/L]were lower than those in control group [(0.49 ±0.12),(7.89 ±1.35)](P<0.05).The total efficacy in observation group was higher than that in control group (70.0%vs.45.0%,P<0.05).Conclusion Shuxuening injection joint epalrestat has the exact efficacy in treatment of patients with diabetic nephropathy.

Mechanism of tumor dormancy / 国际肿瘤学杂志

Tumor dormancy is a phase of occurrence and progress of cancer.In recent years,with the further study of circulating tumor cells,disseminated tumor cells and cancer stem cells,it is found that they are closely related to the tumor dormancy.Moreover,tumor microenvironment is another important factor in tumor dormancy.

Efficacy analysis of laparoscopic adjustable gastric placation for obesity / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To analyze the efficacy and safety of laparoscopic adjustable gastric placation (LAGBP), a new procedure for surgical treatment of obesity.</p><p><b>METHODS</b>Clinical and 1-year follow-up data of 10 patients who underwent LAGBP in our department between September and November 2011 were analyzed retrospectively.</p><p><b>RESULTS</b>The mean operative time was (93.0±13.4) min, while the mean intraoperative blood loss was (15.5±4.7) ml. The mean excessive body weight loss rate(%EWL) at 3, 6, 9 and 12 months after the operation was 25.1%, 40.6%, 45.3% and 50.8% respectively. There were no severe post operative complications.</p><p><b>CONCLUSIONS</b>LAGBP is associated with high safety and good short-term efficacy.</p>

Study on the isoniazid induced cellular damage and the expressions of Fas/Fas ligand of HepG2 / 中华传染病杂志

Objective To establish a model of isoniazid induced necrosis and apoptosis in HepG2 cell and to observe the expressions of Fas/Fas ligand (FasL) in this model.Methods HepG2 cells were treated with different dosages of isoniazid (0,1,2,4,6 and 8 mg/mL) or blank control for 24 hours.Flow cytometer was used to observe the cellular morphology of the HepG2 cell.Annexin V/propidium iodide double staining and flow cytometry were employed to detect the necrosis and apoptosis of HepG2 cells.The expressions of Fas/FasL on the cells were also determined by flow cytometry.The data were analyzed by one-way ANOVA.The comparisons between the drug groups and the control group were performed by using Dunnett t test. Results The higher the dose of isoniazid (4,6,8 mg/mL) was,the more necrosis and apoptosis were observed.In the 4,6 and 8mg/mL isoniazid arms,the total mortality rates were all higher than the control group [(32.1 ±7.5)％,(34.9±8.1)％,(38.2±9.4)％ vs (7.2±1.5)％ respectively](t=4.62,5.14 and 5.75,respectively; all P＜0.01 ).The expression levels of Fas increased along with the dose of isoniazid increasing [(8.7±2.2)％,(11.5±2.8)％,(12.3±3.0)％ and (10.6±2.9)％ in isoniazid 2,4,6and 8 mg/mL arms,respectively],which were all higher than that in control arm [(3.1 ±0.8) ％](t=2.97,P＜0.05; t=4.46,P＜0.01; t=4.88,P＜0.01; t=3.98,P＜0.05).Furthermore,the expressions of FasL increased as well when the dose of isoniazid increased.The expression levels of FasL were (16.2±3.5)％,(21.7±4.8)％ and (18.7±4.9)％,respectively in isoniazid 4,6 and 8 mg/mL arms,which were all higher than that in the control group [(7.4±1.4)％](t=3.11,P＜0.01; t=5.06,P＜0.01; t=3.99,P＜0.05).HepG2 cell necrosis increased with isoniazid of 8 mg/mL.However,the increase of apoptosis was not observed.Conclusion Isoniazid can induce HepG2 cell necrosis and apoptosis,and the apoptosis may be related with the increased expressions of the Fas/FasL on the cells.