Alendronate combined with Lactobacillus rhamnosus to combat bone loss in ovariectomized mice / 中华创伤骨科杂志

Objective:To study the protective effect of alendronate combined with Lactobacillus rhamnosus on bone loss in ovariectomized mice.Methods:Fifty female C57BL/6 mice were divided into 5 equal groups ( n=10). Ovariotomy was performed in groups A, B, C and D while a sham operation was performed in group E. Group A was subjected to combined administration of alendronate and Lactobacillus rhamnosus, group B to administration of alendronate, group C to administration of Lactobacillus rhamnosus, and groups D and E to administration of physiological saline only. At 3 months after operation, all the mice were sacrificed to harvest their femurs. Micro CT scanning was performed to detect the bone mineral density (BMD), trabecular relative volume, bone surface area/bone volume, and trabecular thickness and number of trabecular bone. Three-point bending test was used to detect the maximum load, stiffness, ultimate load, Young＇s modulus, and fracture energy. Osteocalcin and alkaline phosphatase levels were measured using blood samples from the mice eyeballs. The 2 groups were compared in terms of all the above indexes. Results:The BMD [(669.87±67.87) mg/cm 3], maximum load [(14.35±0.75) N] and fracture energy [(1,497.43±38.29) J/m 2] in group A were significantly higher than those in group B [(520.07±9.01) mg/cm 3, (11.94±0.82) N and(1,277.61±35.12) J/m 2] and group C [(388.15±25.61) mg/cm 3, (11.10±0.93) N and (1,115.27±63.24) J/m 2] (all P<0.05). The osteocalcin level in group A [(22.25±1.78) ng/mL] was significantly higher than that in group B [(19.08±1.45) ng/mL] and group D [(19.33±1.66) ng/mL] (both P<0.05). The alkaline phosphatase level in group A [(83.21±9.69) ng/mL] was significantly lower than that in group C [(113.16±14.44) ng/mL] and group D [(137.96±14.01) g/mL] (both P<0.05). Conclusion:Alendronate combined with Lactobacillus rhamnosus may play a synergistic role in prevention of bone loss in ovariectomized mice, because combined administration of the two is more effective than administration of either of the two.

miR-7/TGF-β2 axis sustains acidic tumor microenvironment-induced lung cancer metastasis

Acidosis, regardless of hypoxia involvement, is recognized as a chronic and harsh tumor microenvironment (TME) that educates malignant cells to thrive and metastasize. Although overwhelming evidence supports an acidic environment as a driver or ubiquitous hallmark of cancer progression, the unrevealed core mechanisms underlying the direct effect of acidification on tumorigenesis have hindered the discovery of novel therapeutic targets and clinical therapy. Here, chemical-induced and transgenic mouse models for colon, liver and lung cancer were established, respectively. miR-7 and TGF-β2 expressions were examined in clinical tissues (n = 184). RNA-seq, miRNA-seq, proteomics, biosynthesis analyses and functional studies were performed to validate the mechanisms involved in the acidic TME-induced lung cancer metastasis. Our data show that lung cancer is sensitive to the increased acidification of TME, and acidic TME-induced lung cancer metastasis via inhibition of miR-7-5p. TGF-β2 is a direct target of miR-7-5p. The reduced expression of miR-7-5p subsequently increases the expression of TGF-β2 which enhances the metastatic potential of the lung cancer. Indeed, overexpression of miR-7-5p reduces the acidic pH-enhanced lung cancer metastasis. Furthermore, the human lung tumor samples also show a reduced miR-7-5p expression but an elevated level of activated TGF-β2; the expressions of both miR-7-5p and TGF-β2 are correlated with patients' survival. We are the first to identify the role of the miR-7/TGF-β2 axis in acidic pH-enhanced lung cancer metastasis. Our study not only delineates how acidification directly affects tumorigenesis, but also suggests miR-7 is a novel reliable biomarker for acidic TME and a novel therapeutic target for non-small cell lung cancer (NSCLC) treatment. Our study opens an avenue to explore the pH-sensitive subcellular components as novel therapeutic targets for cancer treatment.

Porphyromonas gingivalis exacerbates ulcerative colitis via Porphyromonas gingivalis peptidylarginine deiminase / 国际口腔科学杂志·英文版

Ulcerative Colitis (UC) has been reported to be related to Porphyromonas gingivalis (P. gingivalis). Porphyromonas gingivalis peptidylarginine deiminase (PPAD), a virulence factor released by P. gingivalis, is known to induce inflammatory responses. To explore the pathological relationships between PPAD and UC, we used homologous recombination technology to construct a P. gingivalis strain in which the PPAD gene was deleted (Δppad) and a Δppad strain in which the PPAD gene was restored (comΔppad). C57BL/6 mice were orally gavaged with saline, P. gingivalis, Δppad, or comΔppad twice a week for the entire 40 days (days 0-40), and then, UC was induced by dextran sodium sulfate (DSS) solution for 10 days (days 31-40). P. gingivalis and comΔppad exacerbated DDS-induced colitis, which was determined by assessing the parameters of colon length, disease activity index, and histological activity index, but Δppad failed to exacerbate DDS-induced colitis. Flow cytometry and ELISA revealed that compared with Δppad, P. gingivalis, and comΔppad increased T helper 17 (Th17) cell numbers and interleukin (IL)-17 production but decreased regulatory T cells (Tregs) numbers and IL-10 production in the spleens of mice with UC. We also cocultured P. gingivalis, Δppad, or comΔppad with T lymphocytes in vitro and found that P. gingivalis and comΔppad significantly increased Th17 cell numbers and decreased Treg cell numbers. Immunofluorescence staining of colon tissue paraffin sections also confirmed these results. The results suggested that P. gingivalis exacerbated the severity of UC in part via PPAD.

Heparin-induced thrombocytopenia is associated with a high risk of mortality in critical COVID-19 patients receiving heparin-involved treatment

BackgroundCoronavirus infectious disease 2019 (COVID-19) has developed into a global pandemic. It is essential to investigate the clinical characteristics of COVID-19 and uncover potential risk factors for severe disease to reduce the overall mortality rate of COVID-19. MethodsSixty-one critical COVID-19 patients admitted to the intensive care unit (ICU) and 93 severe non-ICU patients at Huoshenshan Hospital (Wuhan, China) were included in this study. Medical records, including demographic, platelet counts, heparin-involved treatments, heparin-induced thrombocytopenia-(HIT) related laboratory tests, and fatal outcomes of COVID-19 patients were analyzed and compared between survivors and nonsurvivors. FindingsSixty-one critical COVID-19 patients treated in ICU included 15 survivors and 46 nonsurvivors. Forty-one percent of them (25/61) had severe thrombocytopenia, with a platelet count (PLT) less than 50x109/L, of whom 76% (19/25) had a platelet decrease of >50% compared to baseline; 96% of these patients (24/25) had a fatal outcome. Among the 46 nonsurvivors, 52{middle dot}2% (24/46) had severe thrombocytopenia, compared to 6{middle dot}7% (1/15) among survivors. Moreover, continuous renal replacement therapy (CRRT) could induce a significant decrease in PLT in 81{middle dot}3% of critical CRRT patients (13/16), resulting in a fatal outcome. In addition, a high level of anti-heparin-PF4 antibodies, a marker of HIT, was observed in most ICU patients. Surprisingly, HIT occurred not only in patients with heparin exposure, such as CRRT, but also in heparin-naive patients, suggesting that spontaneous HIT may occur in COVID-19. InterpretationAnti-heparin-PF4 antibodies are induced in critical COVID-19 patients, resulting in a progressive platelet decrease. Exposure to a high dose of heparin may trigger further severe thrombocytopenia with a fatal outcome. An alternative anticoagulant other than heparin should be used to treat COVID-19 patients in critical condition. FundingThis investigation was supported by grants 2016CB02400 and 2017YFC1201103 from the National Major Research and Development Program of China.

Diagnostic value of magnetic resonance imaging conventional sequences and diffusion weighted imaging in polycystic ovary syndrome / 中华内分泌外科杂志

Objective To investigate the diagnostic value of magnetic resonance imaging(MRI) conventional sequences and diffusion weighted imaging (DWI) in polycystic ovary syndrome(PCOS).Methods 75 patients with clinical-confirmed PCOS and 46 healthy women in Yuyao Traditional Chinese Medicine Hospital from Oct.2016 to Apr.2018 were selected.All the subjects received MRI conventional sequences and DWI examination.The size of ovary and number of follicles were observed,and apparent diffusion coefficient (ADC) values in ovarian stroma were measured.The ovarian morphologic changes and ADC values between the two groups were compared.Results The volume of ovary and number of follicles in the control group were (6.6±1.5)cm3 and 6.9±1.8,the difference was statistically significant compared with that in PCOS group [(11.8±2.8)cm3,13.6±3.6)](P＜0.05).ADC value in ovarian stroma in PCOS group was reduced,and the difference was also statistically significant compared with that in the control group(P＜0.05).The sensitivity and specificity of diagnosing PCOS with ADC threshold of 1.39×10-3 mm2/s were 82.2％ and 68.9％,respectively.Conclusion MRI conventional sequences and DWI are helpful in diagnosis of PCOS,which has important significance for the clinical treatment and prognosis evaluation.

Continuing professional development for general practitioners after residency training in Shanghai Weifang community health service center / 中华全科医师杂志

A continuing professional development program was implemented in Shanghai Weifang Community Health Center for general practitioners (GPs) who completed residency training.The program consisted of four aspects: clinical ability, scientific research ability, teaching ability, and management ability.Clinical ability-building included the re-practice of specialty skills, expert mentoring in clinics, continuing education courses and clinic skills competitions.The scientific research ability was built from participating in research projects and talent cultivation programs.GPs were encouraged to be clinical mentors, and teaching ability was developed through attending teacher-training class and teaching practice.For those who were interested in management, the administrative positions related to medical, teaching and research would be provided.The implementation of the program received strong policy support from the health administrations at various levels.

Meta-analysis of posterior laminectomy and instrumented fusion versus laminoplasty in treatment of multilevel cervical spondylotic myelopathy / 中国组织工程研究

BACKGROUND:Many studies concern the comparison of posterior laminectomy and instrumented fusion and posterior laminoplasty for multilevel cervical spondylotic myelopathy, but the sample size of many studies has limitations. There is lack of objective evaluation on advantages and disadvantages of two surgical methods. OBJECTIVE:To compare the efficacy and safety of posterior laminectomy and instrumented fusion and laminoplasty in the treatment of multilevel cervical spondylotic myelopathy. METHODS:A systematic search of al the studies published was conducted on the PubMed, Cochrane Central, EMbase, the ISI Web of Knowledge Database, CMB, CNKI, VIP and Wanfang databases. Randomized and non-randomized control ed trials that compared between posterior laminectomy and instrumented fusion and laminoplasty for multilevel cervical spondylotic myelopathy were identified. Meta-analyses were performed in postoperative Japanese Orthopaedic Association scores, cervical range of motion, cervical curvature index, incidence of C5 nerve root paralysis and incidence of axial symptoms. RESULTS AND CONCLUSION:(1) Fourteen studies involving 1 024 patients were included. Among the patients, 519 underwent laminectomy and instrumented fusion and 505 underwent laminoplasty. (2) The results of the meta-analysis indicated that, compared with laminectomy and instrumented fusion group, laminoplasty group had advantages of a lower incidence of C5 palsy [RR=2.24, 95%CI(1.33,3.75), Z=3.05, P<0.05] and smal degree of cervical rotation injury [SMD=-0.71, 95%CI(-2.21,-1.2), Z=6.63, P<0.05]. However, the two groups had no statistical difference in postoperative Japanese Orthopaedic Association score, cervical curvature index and the incidence of axial symptoms. (3) These results suggested that both laminectomy and instrumented fusion and laminoplasty were demonstrated to be effective for multilevel cervical spondylotic myelopathy. Laminoplasty had obvious advantages of decreasing the degree of cervical rotation injury and lowering incidence of C5 palsy. However, in the process of clinical diagnosis and treatment, the patient’s condition should be combined. The long-term clinical efficacy of the technology needs more clinical work to confirm.

Effect of conversion to enteric-coated mycophenolate sodium on gastrointestine-related quality of life after renal transplantation / 中华器官移植杂志

Objective To evaluate the effect of conversion from mycophenolat mofetil (MMF) to enteric-coated mycophenolate sodium (EC-MPS) on gastrointestine-related quality of life,as well as the safety and efficacy.Method A total of 41 renal transplant (RT) recipients were converted from MMF to EC-MPS (46.3 ± 17.1) months after the operation due to the gastrointestinal side effects of MMF,with a mean time of 46.3 months.Before the conversion and 12 weeks later,the patients were evaluated with Gastrointestinal Quality of Life Index (GIQLI) questionnaire,and the safety and efficacy were assessed.Result The average dose of MMF was 846.7 ± 291.3 mg/day before the conversion,with a mean dose of 639.5 ± 186.4 mg/day for EC-MPS.The total score of GIQLI was 103.6 ± 10.7 before the conversion,and 12 weeks after conversion 118.3 ± 15.1,with a statistical significance (P＜0.05).The safety of EC-MPS was excellent without infection,acute rejection episode,loss of allograft or death.The serum creatine was 136.9 ± 35.7 mol/L before conversion and 128.4± 40.8 mol/L after conversion (P ＞ 0.05).Conclusion For the RT patients with gastrointestinal side effects of MMF,conversion to EC-MPS could significantly alleviate gastrointestinal illness,and improve quality of life,with excellent safety and efficacy.

Digital gene expression profiling analysis of the early adventitious shoot formation in Arabidopsis thaliana / 生物工程学报

Most current research in the field of adventitious shoot formation is focused on the regulatory function of a single gene. However, a systematic transcriptomic analysis of the early adventitious shoot formation is still lacking. Here, we analyzed the transcriptome profiling of the early adventitious shoot formation in Arabidopsis by RNA-seq high throughput sequencing technology, and identified 2 457 differentially expressed genes. Detailed categorization revealed that these genes were mainly involved in hormone homeostasis or signal transduction, callus and lateral root formation, shoot apical meristem development and photosynthesis. Further pathway enrichment analysis showed that genes involved in phenylalanine metabolism and phenylpropanoid biosynthesis were significantly enriched. Moreover, exogenous phenylalanine could repress adventitious shoot formation, indicating that phenylalanine metabolism and phenylpropanoid biosynthesis might be important for adventitious shoot formation.

Association of genetic polymorphisms of transcription factor 7-Like 2 gene and post-transplantation diabetes mellitus in Han Zhejiang population / 中华器官移植杂志

Objective To study the relationship between rs290487,rs7903146 of transcription factor 7-Like 2 (TCF7L2) gene and post-transplantation diabetes mellitus in Han Zhejiang population.Method We genotyped two single nucleotide polymorphisms (SNPs) across the TCF7L2 gene in 90 unrelated post-transplantation type 2 diabetes mellitus (PTDM) patients,112 unrelated non-PTDM patients,and a set of post-transplantation diabetes mellitus patients (n =68).Genotyping was performed using direct sequencing SNP Genotyping Assays.The association of SNPs with post-transplantation diabetes mellitus was analyzed.Result In this study,there was statistically significant difference in the T-allele of TCF7L2.rs7903146 between PTDM group (5.1％) and non-PTDM group (1.3％) (P＜0.05).For rs7903146,the frequencies of genotype C/C,C/T (70.0％) and T/T (35.8％) was statistically significant in PTDM group (P＜0.05).For rs290487,the frequencies of genotype C/C,C/T and T/T was 14.7％,38.2％ and 47.1％ respectively in PTDM group,P＞ 0.05.The incidence of PTDM was significantly higher in patients with the CT genotype (odds ratio 18.54 [95％ CI 1.21-282.26],P =0.03).Conclusion With the current sample size,we found that the CT genotype of rs7903146 was significantly associated with post-transplantation diabetes mellitus.

Percutaneous antegrade ureteral stent in the treatment of renal graft ureteral obstruction: 11eases report / 中华器官移植杂志

ObjectiveTo explore the efficacy and safety of percutaneous antegrade stenting in the treatment of ureteral obstruction after renal transplantation.MethodsWe retrospectively reviewed 11patients with renal graft ureteral obstruction (2 cases of acute obstruction and 9 cases of chronic obstruction) from March 2009 to March 2011.The etiology of the obstruction was renal graft-ureter-bladder anastomotic stricture in 5 cases,stone obstruction in 2 cases,and undetermined in 4 cases.Renal graft and collecting system were examined by ultrasonography preoperatively to select suitable puncture position,and then ureteropyelography was performed under X-ray guidance.When the obstruction location was clear,the urology guidewire was implanted to the bladder by needle,and then guidewire was released by cystoscopy.Ureteral stent was implanted along the guidewire,and upper ureteral stents was observed under X-ray. After removal of guidewire,the stent location was confirmed once again.The renal pelvis fistula drainage lasted for 1-2 weeks,and ureteral stent to 6 months to one year.Ultrasound and renal function were tested after 1week,1month,3 months and 6 months,and then every six months.ResultsOperation was done successfully in 10 patients,and failed in one case due to a long segment of ureteral stenosis.The operating time of ureteral stent implantation was 54±27 min.Serum creatinine of patients was reduced from preoperative 326±147 to postoperative 89±49 μmol/L.During a follow-up period of 6 to 27 months,no complications occurred.ConclusionPercutaneous antegrade stenting in the treatment of ureteral obstruction after renal transplantation is safe and effective.

Correlation between post-transplant glomerular filtration rate in 1 year and long-term graft survival in renal transplant patients / 中华器官移植杂志

Objective To explore the correlation between post-transplant glomerular filtration rate (GFR) in 1 year and long-term graft survival in renal transplant patients.Methods The clinical data of 334 patients who received their cadaveric kidney transplantations between November 1994 and October 2004 were analyzed retrospectively.According to the GFR at one year after transplant operation, normal GFR group was defined as GFR more than or equal to 1.083 ml/s, while patients whose GFR less than 1.083 ml/s were fallen into abnormal GFR group.Cockeroft-Gault (C-G) formula was used to compare the difference in the renal function between the two groups.Kaplan-Meier assay was used to compare the difference in the allograft survival between the two groups in the functional renal allograft or the non-functional renal allograft.The correlativity of GFR level at the first year and the GFR level at the 5th year was analyzed.Results The GFR level at the first year after transplantation was proportional to the graft survival time of the kidney.Five and ten years after transplantation, the renal transplantation long-term survival rate in the normal renal function groups was significantly higher than in the abnormal renal function groups (P＜0.05).As compared with the GFR level at the first year after transplantation, the changes in amplitude of GFR level at the 5th year after transplantation was (0.080 ±0.248) ml/s, and the descent had a positive correlation with GFR level at the 5th year after transplantatioa Conclusion GFR level at the first year after transplantation predicts long-term renal functioa The higher of GFR level at the first year, the higher of GFR level at the 5th year.

Dynamic simulation and analysis of ERK signal pathway activated by epidermal growth factor receptor / 生物医学工程学杂志

Extra-cellular signal regulated kinase (ERK) signal transduction is an important signaling pathway, involved in a variety of processes in cell and is closely related with the occurrence of human cancers. By the application of dynamic method, the ERK signal pathway activated by epidermal growth factor receptor (EGFR) was simulated and analyzed in this work. The influence of the mutation of adaptor protein on the expression of phosphorylated ERK signal was analyzed, and the complex coupling of the protein kinases and their kinetic function were analyzed. The obtained results are helpful to understanding the mechanism of tumor development and to finding out new target molecules in the ERK signaling system.

Establishment of pTet-on rat precartilaginons stem cell strains and construction of pTRE-PTHrP(1-36) responsive plasmids / 中华创伤杂志

Objective To establish a doxycycline-controlled immortalized pre-cartilaginons stem cells (IPCSCs) strains, clone parathroid hormone-related peptide[PTHrP(1-36)] gene and construct re- sponsive plasmid, pTRE-PTHrP (1-36). Methods Plasmid pTet-on was transfected into IPCSCs by using LipoinfectaminTM 2000 and then the stable clones were obtained by G418 screening. The doxycyc- line was added into the medium of monoclonal cells that were transiently transfected with plasmid pTRE- 2Hyg-Lue. The total RNA was extracted from PCSCs and the PTHrP(1-36) gene obtained by RT-PCR method. Then, the PTHrP (1-36) gene was subcloned to plasmids of Tet-responsive element with the se- lection marker of hygromycin pTRE-2Hyg to construct recombinant eukaryotic expression plasmid pTRE- PTHrP(1-36). After transferred into E. coli-DH5α, the clone was amplified, the recombinant plasm0ids were purified and identified by double-enzyme digestion. Results The doxycycline induced IPCSCs line was obtained, with 50 times higher than the non-induced cell line. Double enzyme digestion analysis and sequencing showed that the target gene was cloned into recombinant plasmid. Conclusions The induced IPCSCs line can be used to highly express alien genes. The responsive plasmid containing PTHrP (1-36) gene may be premising for rigorous control of PTHrp (1-36) gene expression.

Separation,identification and immortalization of precartilaginous stem cells from neonatal rats / 中国组织工程研究

BACKGROUND: Precartilaginous stem cells (PSCs) have strong proliferation ability and differentiation potential,but they are instable and prone to differentiate.Importing exogenous gene could immortalize them and leave phenotype character unchanged.OBJECTIVE: To establish immortalized precartilaginous stem cells (PSCs) from neonatal SD rats in vitro for the further related research about the differentiation mechanism and clinical application of precartilaginous stem cells.DESIGN,TIME AND SETTING: Single sample observation.The study was carried out in the Department of Orthopedics.Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from October 2005 to September 2006.MATERIALS: Neonatal SD rats,irrespective of gender,24-hour old,were used for prepare PSCs.METHODS: By using LipofectamineTM 2000,a gene transfection reagent,plasmid pCMVSV40T/PUR containing the simian virus 40 large T antigene gene (SV40Tag) was transfected into the primary cultured PSCs isolated by immuniomagnetic beads coasted with the second antibody.Colonies were isolated by puromycin selection and expanded by many passages.MAIN OUTCOME MEASURES: Biological character of PSCs; plasmid identification; biological character of transfected cells and identification; RT-PCR; growth curve.RESULTS: Immunomagnetic beads separation system obtains PSCs,which was confirmed as fibroblast growth factor receptor-3 (FGFR-3) positive PSCs.Double restriction enzyme was cut,electrophoresis confirmed pCMV was 3 kb,SV40T was 2.3 kb.A particular anti-puromycin cell clone was acquired,which was confirmed as FGFR-3 positive PSCs.The total RNA was isolated from the positive cell clones,and a 588 bp fragment,which was specific for the SV40T antigene gene,was amplified.The transfected cells were expanded to immortalized cell strain,named as immortalized precartilaginous stem cells (IPSCs).Thepopulation doubling time of IPSCs was (22.98±2.77) hours,no significant effect of subculture,freezing and recovering had been found.CONCLUSION: Precartilaginous stem cells could be isolated from neonatal SD rats,cultured in vitro,and immortalized through the transfection of pCMVSV40T/PUR.

Research progress in the quantitative structure-antitumor activity relationship studies of taxol derivatives / 中国药理学通报

The up-to-date quantitative structure-antitumor activity relationship studies of taxol analogues based on molecular modeling techniques were reviewed in the present paper. Both the common understandings and the debates about the contributions of several key structural groups, including the C13 side chain, the substituents of C14, C2 and C10 as well as the oxtane ring to the biological activities of taxol derivatives were summerized. In addition, the studies about other factors influencing the activity except for the structure and the classification research of the molecules were also discussed.