Effect of warm-needling moxibustion on anterior cruciate ligament injury repair and related growth factors in rabbits with knee osteoarthritis / 中国组织工程研究

BACKGROUND:Warm-needling moxibustion can effectively treat knee osteoarthritis.Degeneration,injury and fracture of the anterior cruciate ligament can affect the local stability of the knee joint,and then induce the formation of knee osteoarthritis.Whether warm-needling moxibustion can repair the injured cruciate ligament and the mechanism of action are still unclear. OBJECTIVE:To observe the effects of warm-needling moxibustion on the morphology of the anterior cruciate ligament and the expression of insulin growth factor-1 and transforming growth factor-β in rabbits with knee osteoarthritis and to clarify the mechanism of anterior cruciate ligament repair by warm-needling moxibustion. METHODS:Thirty New Zealand rabbits were randomly divided into blank group,model group and warm-needling moxibustion group,with 10 rabbits in each group.Knee osteoarthritis model was established by plaster cast immobilization.The blank group was not intervened.Rabbits in the model group rabbits were fixed in a rabbit holder for 15 minutes every day.The warm-needling moxibustion group was treated with warm acupuncture,once a day,7 days as a course of treatment,a total of two courses.After treatment,the imaging changes of the anterior cruciate ligament were observed by MRI and MRI grading statistics were performed.Morphological changes of the anterior cruciate ligament were observed by transmission electron microscope and hematoxylin-eosin staining.mRNA and protein expressions of insulin growth factor-1 and transforming growth factor-β were detected by RT-PCR and western blot,respectively. RESULTS AND CONCLUSION:MRI examination:Compared with the blank control group,the anterior cruciate ligament in the model group was thickened,edematous,and partially torn,and the difference in grading statistics was statistically significant(P＜0.05).Compared with the model group,the anterior cruciate ligament in the warm-needling moxibustion group was slightly thickened,with mild edema and no tearing,and the difference in grading statistics was statistically significant(P＜0.05).General observation:In the model group,the surface of the anterior cruciate ligament was glossy and faded,with the edge being covered with flocculent periosteum and obvious tissue necrosis;in the warm-needling moxibustion group,the surface of the ligament was glossy,and the ligament was in a normal helical shape.Hematoxylin-eosin staining:In the model group,there was obvious tissue necrosis in the anterior cruciate ligament,a large number of new capillaries,loosely arranged fibroblasts and collagen fibers.In the warm-needling moxibustion group,there was a small amount of tissue necrosis and few new vessels in the anterior cruciate ligament,and the cells and collagen fibers were loosely and irregularly arranged.Transmission electron microscopy:In the model group,the fibers in the anterior cruciate ligament were arranged in a disordered way with uneven thickness and distribution,and there are more fibroblasts that were irregular in morphology;in the warm acupuncture group,the fibers were basically arranged longitudinally,with uneven thickness and distribution,and a small number of oval-shaped fibroblasts were observed.RT-PCR and western blot assay:mRNA and protein expressions of insulin growth factor-1 and transforming growth factor-β were significantly decreased in the model group compared with the blank control group(P＜0.05),but significant increased after treatment with warm-needling moxibustion(P＜0.05).To conclude,warm-needling moxibustion can alleviate anterior cruciate ligament injury and regulate the expression of insulin growth factor-1 and transforming growth factor-β to treat knee osteoarthritis.

Expression and clinicopathologic features of ARL5B in esophageal cancer and its mechanism / 西安交通大学学报(医学版)

【Objective】 To evaluate the clinical implications of ARL5B in esophageal cancer and its underlying mechanisms by using bioinformatics methods. 【Methods】 ARL5B transcriptomic expression data were obtained from The Cancer Genome Atlas (TCGA), R software was employed to detect the differential expression mRNAs, and related clinical information was collected for survival analysis. To validate the bioinformatics results, Real-time quantitative PCR (qRT-PCR) and Western blotting were carried out for clinical specimens of esophageal cancer tumor tissues and adjacent tissues. Immunohistochemistry was used to evaluate the expression of ARL5B and its associated clinicopathologic features. The underlying mechanisms of ARL5B in esophageal cancer were preliminarily explored by bioinformatics and qRT-PCR. 【Results】 Bioinformatics method showed that the expression of ARL5B in human esophageal cancer tissues was significantly higher than in adjacent tissues and correlated with poor prognosis. Clinical specimens were detected, the expressions of ARL5B mRNA and protein were the highest in metastases lymph node, followed by esophageal cancer tissues and adjacent tissues, which corresponded with bioinformatics results. The expression of ARL5B was strongly correlated with lymph node metastases and advanced clinical stage. Kaplan-Meier analysis results denoted high ARL5B level, indicating poor prognosis. Enrichment analysis showed that ARL5B was associated the biological processes such as vacuolar transport, late endosome to lysosome transport, and organelle localization. Protein-protein interaction analysis (PPI) suggested that ARL5B might interact with VPS16, KIF1A and TOM1, whose expressions were verified by qRT-PCR and positively correlated with ARL5B expression. 【Conclusion】 ARL5B was highly expressed in esophageal cancer and associated with lymph node metastases, advanced clinical stage, and poor prognosis. ARL5B may be involved in the progression of esophageal cancer with several molecular mechanisms.

Research on clinical effectiveness of fuzzy failure mode and effects analysis combined with multi-dimensional management in orthopedic rehabilitation equipment management / 中国医学装备

Objective:To construct a fuzzy failure mode and effects analysis(FMEA)combined multi-dimensional management mode to improve the management quality of orthopedic rehabilitation treatment equipment.Methods:14 types of failure modes in 5 aspects and 25 common situations in 7 aspects of failure causes of orthopedic rehabilitation equipment were selected as risk control objects.Fuzzy algorithm was used to evaluate the risk severity,probability of occurrence,detectability and priority value,and multi-dimensional management countermeasures were developed from the aspects of daily inspection,quality detection,maintenance and emergency management.65 units of rehabilitation treatment equipment in clinical use in the hospital were selected and divided into control group(49 units)and observation group(54 units,including 38 units in the control group and 16 newly added units)according to different management methods.The control group adopted traditional FMEA priority value management method,and the observation group adopted fuzzy FMEA combined multi-dimensional management method.The equipment failure ratio,FMEA risk assessment value and medical equipment service quality were compared between the two groups.Results:The proportion of failures due to abnormal power-on self-test,poor treatment operation quality,malfunction shutdown,unqualified quality inspection and external damage of the orthopedic rehabilitation treatment equipment in the observation group were(0.76±0.89)%,(1.72±1.35)%,(1.07±1.22)%,(2.19±0.93)%and(0.13±0.26)%,respectively,which was lower than that of control group,the difference was statistically significant(Z=2.455,Z=2.833,Z=2.236,Z=3.637,Z=2.014;P<0.05).The severity,incidence,detectability and priority values of orthopedic rehabilitation equipment risk in the observation group were(3.34±1.57)points,(2.56±0.94)points,(3.63±1.20)points and(31.29±20.80)points,respectively,which were lower than that in the control group,the difference was statistically significant(Z=2.117,Z=2.665,Z=3.149,Z=3.466;P<0.05).The results of 50 randomly selected patients,37 related medical staff and 120 random inspection showed that the patient satisfaction,medical staff recognition and random inspection standardization of the equipment in the observation group were 88%(44/50),97.30%(36/37)and 96.67%(116/120),respectively,which were higher than those in the control group,the difference was statistically significant(x2=4.882,x2=6.198,x2=8.819;P<0.05).Conclusion:Fuzzy FMEA combined with multi-dimensional management mode can reduce the failure rate of orthopedic rehabilitation equipment,control the severity,occurrence probability and detectability of equipment risks,and improve the equipment operation safety and clinical service level.

3-Methyladenine alleviates extracellular matrix deposition in early diabetic nephropathy by inhibiting VEGF signaling / 中华内分泌代谢杂志

Objective:To investigate the effects of 3-methyladenine(3-MA)on extracellular matrix deposition in early diabetic nephropathy(DN)and its mechanism.Methods:A streptozotocin(STZ)-induced type 1 diabetes mouse model was used, and the mice were divided into vehicle control group, diabetes group(STZ group), 3-MA group, and chloroquine(CQ)group, 8 mice in each group. After 6 weeks of intervention, both kidneys were harvested, and the kidney-to-body weight ratio was recorded. Western blotting was performed to detect protein expressions of renal cortex fibronectin, α-smooth muscle actin(α-SMA), LC3, Beclin 1, p62, and vascular endothelial growth factor(VEGF). Immunohistochemistry was used to observe kidney fibronectin staining. Bioinformatics analysis was conducted on shared genes between diabetic nephropathy(DN)gene targets and 3-MA predicted gene targets.Results:Both 3-MA and CQ exhibited certain hypoglycemic effects in diabetic mice. Compared to the STZ group, the kidney-to-body weight ratio decreased in the 3-MA group( P<0.05). Western blotting showed that 3-MA reduced the expression of renal cortex matrix-related proteins fibronectin and α-SMA in diabetic mice( P<0.05 or P<0.01). Immunohistochemistry also revealed that 3-MA reduced fibronectin staining in the kidneys of diabetic mice. Both 3-MA and CQ inhibited the protein expression of renal cortex Beclin 1 in diabetic mice(both P<0.05), while 3-MA increased the expression of renal cortex p62( P<0.05). Bioinformatics analysis indicated a connection between shared genes of DN gene targets and 3-MA predicted gene targets with the VEGF signaling pathway. Western blotting results further showed that 3-MA reduced renal cortex VEGF expression in diabetic mice( P<0.01). Conclusion:3-MA can alleviate extracellular matrix deposition in the kidneys of early DN mice by inhibiting the VEGF signaling pathway.

Investigation and analysis on pneumoconiosis in migrant workers without liability subject in Hunan Province / 中国职业医学

OBJECTIVE: To analyze the epidemiological characteristics of pneumoconiosis among migrant workers without liability subject(hereinafter referred to as Pneumoconiosis without Liability Subject) in Hunan Province. METHODS: The cases of pneumoconiosis without liability subject from 2017 to 2019 in Hunan Province were selected as the research subjects using typical sampling method. They were clinical diagnosis by occupational disease diagnostic institutions. The distributions of age, gender, length of service, area, type of work, type of pneumoconiosis, pneumoconiosis stage and the situation of poor households with filing and registration card were analyzed. RESULTS: A total of 18 870 cases of pneumoconiosis without liability subject were clinically diagnosed in Hunan Province from 2017 to 2019. The patients were mainly males(accounting for 99.8%), with the age ranged 50-65 years old(64.7%). Most of them had dust exposure service length of 5-29 years(78.4%). The cases of stage Ⅰ, Ⅱ and Ⅲ pneumoconiosis accounted for 32.2%, 26.0% and 41.8% respectively. The main types of disease were coal workers′ pneumoconiosis and silicosis(accounted for 99.3%). The first five geographical distributions were Chenzhou City, Zhuzhou City, Hengyang City, Yiyang City and Shaoyang City, accounting for 17.9%, 14.6%, 14.1%, 11.8% and 9.2% respectively. The distribution of work types were mainly mine-related jobs(91.3%). There were 1 774 cases who had complications(9.4%), of which the top three complications were emphysema, pulmonary and bronchial infection and tuberculosis. There were 3 662 cases with poor households archives and cards(19.4%). CONCLUSION: The hazards of pneumoconiosis among migrant workers in Hunan Province should not be ignored. In 2017, Hunan Province took the lead in launching a large-scale basic medical treatment and rescue operation for migrant workers with pneumoconiosis, which helped solve the problem of pneumoconiosis in migrant workers who had no professional history certification and responsible employer.

Long non-coding RNA SPATA31D5P promoting proliferation, migration and invasion of breast cancer cells through adsorption of microRNA-320a / 解剖学报

Objective Long non-coding RNA(lncRNA) are aberrantly expressed in breast cancer(BC) and strongly associated with its survival prognosis. The aim of this study is to investigate the expression and effect of IncRNA SPATA31D5P on the invasion and migration capacity of breast cancer cells through adsorption of miR-320a. Methods Totally 30 cases of BC tissues and paraneoplastic tissues were collected, and the expression levels of SPATA31D5P in BC tissues and BC cell lines were detected by Real-time PCR. MDA-MB-231 cells were transfected with SPATA31D5P siRNA interference vector, and cell proliferation, invasion and migration capacity were determined using the cell counting kit-8 assay (CCK-8), 5-ethynyl-2'- deoxyuridine(EdU), Transwell and wound-healing assay respectively. And cell cycle and apoptosis were detected by flow cytometry. Bioinformatics approachs were used to screen for miRNAs that could bind complementarily to SPATA31D5P, and the regulatory effect of SPATA31D5P on miR-320a was detected by Real-time PCR and dual luciferase reporter assay. Results SPATA31D5P levels were significantly higher in BC tissues than in adjacent normal breast tissues, and SPATA31D5P expression was higher in each BC cell line than in normal breast epithelial cells MCF10 A. The level of SPATA31D5P in the interference group was 0. 288±0. 052, which was lower than that of the blank control group 1. 114±0. 096 and negative control (NC) group 1. 079±0. 128 (P< 0. 01). The proliferation activity of MDA- MB-231 cells in the interfered group was significantly reduced and apoptotic rate was obviously increased compared to the NC and control groups (P<0. 01) ;the Gj phase block was observed in the interfered group; the scratch healing rate and number of perforated cells in the interference group were (14. 36 ± 1. 75) % and (26±1.52), which were lower than (52. 25± 1.87)% and ( 67. 33 ± 2. 91 ) of the NC group (PcO.Ol). Dual luciferase experiments confirmed that SPATA31D5P could directly regulate miR-320a expression and luciferase activity. Conclusion SPATA31D5P is highly expressed in BC, interfering with SPATA31D5P expression effectively inhibits the proliferation, migration and invasion of MDA-MB-231 cells, and the mechanism may be related to the targeted regulation of miR-320a.

Effect of moxa-cone moxibustion at lung's back- / 中国针灸

OBJECTIVE@#To observe the effect of moxa-cone moxibustion at lung's back-@*METHODS@#Sixty SPF-grade healthy male Balb/c mice were randomly divided into a normal group, a model group, an LY294002 group (LY group), an electroacupuncture (EA) group and a moxibustion group, 12 mice in each group. Asthma model was replicated by using ovalbumin (OVA) sensitization. Except the mice in the normal group, all the mice were intraperitoneally injected with sensitization solution (containing 15 μg of OVA and 30 mg of aluminum hydroxide) on the 1st day, 7th day and 14th day, 0.5 mL per mice; from the 15th day, 1% OVA solution was atomized for 20 min, once a day for 2 weeks; the mice in the normal group was treated with identical operations but with 0.9% sodium chloride solution. The mice in the LY group were treated with injection of LY294002 at tail vein on the 13th day, 14th day and 15th day. At the beginning of the 15th day, The mice in the EA group were treated with EA at "Feishu" (BL 13) and "Zhongfu" (LU 1) with disperse-dense wave, frequency of 2 Hz/20 Hz, intensity of 1 mA, 15 min each time, once a day for 2 weeks. The mice in the moxibustion group was treated with moxa-cone moxibustion at "Feishu" (BL 13) and "Zhongfu" (LU 1) from the 15th day, three moxa-cones per acupoint, once a day for 2 weeks. On the 16th day, 18th day and 22nd day, the incubation period of asthma was recorded. On the 29th day, all the samples were collected. The expressions of IL-17 and IL-10 in serum and bronchoalveolar lavage fluid (BALF) were detected by ELISA method. The pathological changes of lung tissue were observed by HE staining. The percentage of Th17, Treg and Th17/Treg ratio in spleen tissue were detected by flow cytometry method.@*RESULTS@#Compared with the normal group, the incubation period of asthma in the model group was significantly shortened (@*CONCLUSION@#The Th17/Treg is imbalanced in asthmatic body. The moxibustion at lung's back-

Expression of polypeptide N-acetylgalactosaminyltransferase-10 in lung cancer / 解剖学报

Objective The purpose of this study is to investigate the expression of the polypeptide N-acetylgalactosaminyltransferase-10 (ppGalNAc-T10) protein in lung and breast and colon cancer. Methods Immunohistochemistry was used to analyse ppGalNAc-T10 expressions in 110 lung cancer tissues, 100 breast cancer tissues and 110 colon cancer tissues. Meanwhile, different cancer has 10 cases of normal tissues as control. Results The expression of ppGalNAc-T10 in normal lung tissue was higher compared with that in lung cancer organizations, and ppGalNAc-T10 positive expression was found to be significantly associated with invasion depth of tumor (P < 0. 01). PpGalNAc-T10 protein expression in breast and colon cancer was no significantly difference than that in non-tumor breast and colon tissue. Conclusion PpGalNAc-T10 expression is a useful marker in lung cancer.

Analysis of results of ultrasound examination of thyroid nodules in nuclear power workers / 中华劳动卫生职业病杂志

Objective@#To investigate the detection of thyroid nodules and related risk factors in nuclear power workers, and to provide scientific evidence for thyroid protection of nuclear power workers.@*Methods@#In December 2018, select 295 workers of a nuclear power production enterprise and 238 administrative staff of it, and select 250 staff members of a thermal power generation enterprise 70 kilometers away from the nuclear power station to conduct thyroid ultrasound examination and questionnaire survey for single factor. Analysis and further multivariate logistic regression analysis were used to study the risk factors of thyroid nodules in the population.@*Results@#Women and smoking history were independent risk factors for the increased incidence of thyroid nodules in the study population; three shifts work pattern was an independent risk factor for the increased prevalence of thyroid nodules in nuclear power workers (P<0.05) , and other factors such as the history of nuclear exposure had no significant correlation with thyroid nodules (P>0.05) .@*Conclusion@#Nuclear exposure has little effect on the prevalence of thyroid nodules in nuclear power workers.

miR-320 inhibits glycometabolism in colorectal cancer by targeting E2F1 gene / 国际肿瘤学杂志

Objective To study the effect of microRNA-320 (miR-320) targeting E2F1 gene on tumor glycometabolism in colorectal cancer.Methods The miR-320 expression level in colorectal cancer cell lines and cancer tissues was detected using quantitative real-time polymerase chain reaction (qRT-PCR).The binding sites of miR-320 and E2F1 were predicted by bioinformatics.Luciferase assay was used to detect the targeting regulation of miR-320 on E2F1.The relationship between E2F1 and miR-320 was verified in mRNA level and protein level.When the miR-320 in SW480 and LOVO ceils was up-regulated and the E2F1 was down-regulated,the changes of glycometabolism in tumor cells were analyzed using glucose/glucose oxidase kit and lactate test kit.Results The qRT-PCR results showed low expressions of miR-320 in colorectal cancer cell lines and cancer tissues (F =42.327,P ＜ 0.001;t =4.345,P =0.023).Luciferase assay showed that miR-320 could negatively regulate the expression of E2F1 (t =4.716,P =0.042).The expression levels of E2F1 protein and mRNA (t =4.780,P =0.041;t =5.506,P =0.031) confirmed that miR-320 could interact with E2F1 in LOVO and SW480 cells.Overexpression of miR-320 could reduce the contents of glucose (t =5.262,P=0.034;t =21.079,P=0.002) and lactic acid (t =9.609,P=0.011;t =18.582,P=0.003) in the cellular supematant in SW480 and LOVO ceils.Down-regulating the expression of E2F1 at the same time could enhance the inhibitory effect of miR-320 on glucose (t =5.128,P =0.036;t =5.089,P =0.037) and lactic acid (t =8.573,P =0.013;t =13.364,P =0.006).Conclusion E2F1 is the target gene of miR-320,and miR-320 can regulate the glycometabolism of colorectal cancer cells by targeting E2F1 gene.

Construction of sRNA-deficient and-overexpressing strains of Yersinia pestis / 军事医学

Objective To construct small RNA deletion and overexpression strains with a length of less than 100 nt in Yersinia pestis.Methods Deletion mutants of the target sRNAs were constructed by increasing the length of homologous regions.Meanwhile, the high copy plasmid pBAD/HisA was modified into an inducible transcriptional vector as an sRNA-overexpression plasmid by using QuikChange lightning site-directed mutagenesis kit .The presence , size, and transcription-al initiation sites of the indicated sRNA were predicted by transcriptome sequencing , primer extension , and previous stud-ies.The full-length DNA fragments of target sRNAs were transformed into the transcriptional vector .The overexpressing strains of sRNAs were identified by Northern Blot .Results and Conclusion Four sRNAs deletion mutants of sR01, sR02, sR03 and HmsA and three sRNAs overexpression mutants MicF , HmsA and CpxQ were successfully constructed .A method of construction of sRNA deficient and overexpressing strains of Y.pestis has been quickly and efficiently established by λ-Red homologous recombination technology and QuikChange ? lightning site-directed mutagenesis kit.

Recombinant neuregulin-1 beta preserves neural stem cell proliferation through extracellular regulated protein kinases signaling pathway in oxygen and glucose deprivation environment / 中华地方病学杂志

Objective To investigate the influence of recombinant neuregulin-1 beta (rhNRG-1β) on neural stem cell proliferation through extracellular regulated protein kinases (ERK) signaling pathway in oxygen and glucose deprivation (OGD) environment.Methods Neural stem cells were obtained from embryonic brain of mice pregnant for 14-17 d,cultured and identified by immunochemical staining through detection of the indicator nestin using the SABC-FITC (POD)double standard kit.Neural stem cells were divided into three experiment groups (OGD group,control group and OGD + rhNRG-1β group).Control group:identified neural stem cells,2 × 107,were cultured for 3 h in the 24-hole culture plate with DMEM/F12 complete culture medium;OGD group:neural stem cells,2 × 107,were cultured in the 24-hole culture plate deprived glucose DMEM/F12 in a wet airtight container (37 ℃ constant temperature),cells were cultured with mixed gas of nitrogen (950 ml/L) and oxygen (50 ml/L) for 1 h,and then the culture medium was replaced with complete culture medium and cultured for 3 h;OGD + rhNRG-1β group:before OGD intervention,100 μg/L rhNRG-1β was given for 3 h.Neurospheres formation:the three groups of stem cells were dispersed into single cells,1 × 106/ml cells were inoculated to culture plates containing cover slips coated with poly lysine,and cultivated for 7 d,and neurospheres formation of the 3 groups of neural stem cells was observed under microscope,which was aimed to record neural stem cells proliferation changes.Colony formation:the three groups of stem cells,vaccinated in 60 mm in a petri dish,2 × 107 in number,were cultivated in complete culture medium for 24 h.The colony formation of the three groups of cells was observed under microscope,and neural stem cells proliferation changes were observed.Western blotting:the change of phosphorylation ERK (pERK) protein of the three groups of stem cells was determined,and the effect of pERK protein expression regulated by rhNRG-1β in mice neural stem cells proliferation through ERK signaling pathway was observed.Results Microscopically the primary cultured stem cells grew in single or in pairs,in a round shape;neural stem cell proliferated in clumps or colony;neural stem cells expressed the specificity of nestin protein markers with fluorescent yellow-green color.The differences in the aspects of the average diameter of neurospheres and neurospheres quantity in the neural stem cells between groups were statistically significant (F =693.66,1 002.09,all P ＜ 0.01),and among them the neuropheres formation of OGD group was significantly suppressed.Formation quantity and average diameter in OGD group [(88.78 ± 7.14) numbers,(62.12 ± 2.52) μm] were significantly lower than those in the control group [(246.34 ± 8.67) numbers,(128.45 ± 2.33) μm] and those in OGD + rhNRG-1β group [(237.87 ± 6.61) numbers,(118.37 ± 2.71) μm,all P ＜ 0.01].The difference of colony formation rate of neural stem cell was statistically significant (F =132.03,P ＜ 0.01),and among them colony formation of OGD group significantly suppressed.Formation rate in OGD group [(11.65 ± 0.94)％] was significantly lower than that in the control group [(33.23 ± 2.93)％] and that in OGD + rhNRG-1β group [(31.42 ± 2.61)％,all P ＜ 0.01].Western blotting showed that the difference of pERK protein expression of neural stem cells between groups was statistically significant (F =63.76,P ＜ 0.01).Relative expression of the pERK protein in OGD group (0.487 ± 0.072) was significantly lower than that in the control group (1.013 ± 0.112) and that in OGD + rh-NRG-1β group (1.752 ± 0.278,all P ＜ 0.01).Conclusion rhNRG-1β preserves neural stem cell proliferation with phosphorylation ERK protein expression up-regulated in oxygen and glucose deprivation environment.

RNA interference of HERC4 inhibits proliferation, apoptosis and migration of cervical cancer Hela cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the effects of silencing HERC4 on the proliferation, apoptosis, and migration of cervical cancer cell line Hela and the possible molecular mechanisms.</p><p><b>METHODS</b>Three HERC4-specific small interfering RNAs (siRNAs) were transfected into Hela cells, and HERC4 expression in the cells was examined with Western blotting. CCK-8 assay, annexin V-FITC/PI assay, and wound healing assay were used to assess the effect of HERC4 silencing on the proliferation, apoptosis and migration ability of Hela cells. The expression levels of cyclin D1 and Bcl-2 in the cells were detected using Western blotting.</p><p><b>RESULTS</b>Transfection of siRNA-3 resulted in significantly decreased HERC4 protein expression (P<0.01). HERC4 silencing by siRNA-3 markedly suppressed the proliferation and migration of Hela cells, increased the apoptosis rate (P<0.01) and reduced the expression levels of cyclin D1 and Bcl-2 (P<0.01).</p><p><b>CONCLUSION</b>Silencing of HERC4 efficiently inhibits the proliferation, migration, and invasion of Hela cells in vitro, and the underlying mechanisms may involve the down-regulation of cyclin D1 and Bcl-2.</p>

Expression of long noncoding RNA LINC01358 in prostate cancer and its effect on the proliferation and migration of prostate cancer cells / 中华男科学杂志

<p><b>Objective</b>To explore the expression of long noncoding RNA (lncRNA) LINC01358 in prostate cancer (PCa) and its effect on the proliferation and migration of PCa cells.</p><p><b>METHODS</b>The lncRNA array was used to screen differentially expressed lncRNAs in PCa and the corresponding carcinoma-adjacent normal tissues from 3 patients. The expressions of LINC01358 in the primary PCa, metastatic PCa, and carcinoma-adjacent tissues were compared using the PCa dataset of the Memorial Sloan Kettering Cancer Center (MSKCC). The data obtained were validated by determining the expression of LINC01358 in the PCa and carcinoma-adjacent tissues of another 10 patients by quantitative real time PCR (qRT-PCR). The effects of lncRNA LINC01358 on the proliferation of DU145 cells and migration of PCa cells were detected by MTT and Transwell assay, respectively.</p><p><b>RESULTS</b>Totally, 79 differentially expressed lncRNAs in the lncRNA array, 36 highly and the other 43 lowly expressed in the PCa tissue. LINC01358 was up-regulated in the cancerous tissue. According to the MSKCC data, the LINC01358 expression was markedly higher in metastatic PCa (5.81±0.19, n = 19) and primary PCa (5.47±0.04, n = 131) than in the PCa-adjacent tissue (5.15±0.07, n = 29) and significantly correlated with postoperative biochemical relapse of the malignancy (P<0.05). qRT-PCR indicated a remarkably higher expression of LINC01358 in the PCa than in the carcinoma-adjacent tissue (6.02±1.12 vs 3.21±0.21, P<0.05). Transfection of the DU145 cells with siRNA significantly decreased the level of LINC01358 and inhibited the proliferation and migration of the PCa cells.</p><p><b>CONCLUSIONS</b>LINC01358 is highly expressed in the PCa tissue and knockdown of LINC01358 may inhibit the proliferation and migration of PCa cells. LncRNA LINC01358 may be involved in the development and progression of PCa and become an index for the early diagnosis as well as a new target for the gene therapy of the malignancy.</p>

Synthesis and biological evaluation of novel pleuromutilin derivatives with nitrogen-containing heterocycles as antibacterial agents / 药学学报

A series of new pleuromutilins derivatives were designed and synthesized through coupling 2-aminothiazole ring of WL001 with different nitrogen-containing substituted heterocycles in the side chain. Their biological activities were evaluated against both Gram-positive and Gram-negative clinical bacteria in vitro Most new compounds displayed specificity to certain strain of bacteria. Particularly, compounds with saturated nitrogen-containing heterocycles exhibited significant antibacterial activities (0.062 5-8 µg · mL(-1)) superior or similar to those of amoxicillin, tiamulin and levofloxcin. Furthermore, treatment with 15a and 15b having piperidine or morpholine residues also could effectively inhibit Gram-negative bacteria. Therefore, our novel findings may provide a new insight into the design of novel pleuromutilin derivatives and lay the basis for further studies on the treatment of drug-resistance of pathogenic bacteria.

Analysis and synthesis of the related substances of palbociclib / 中国药科大学学报

@#To strengthen the quality control of palbociclib, three related substances were separated from the bulk drug. These substances were identified as 8-cyclopentyl-5-methyl-2-((5-(piperazin-1-yl)pyridin-2-yl)amino)pyrido ［2, 3-d］ pyrimidin-7(8H)-one(A), 8-cyclopentyl-5-methyl-2-((5-(piperazin-1-yl)pyridine-2-yl)amino)-6-vinylpyrido ［2, 3-d］ pyrimidin-7(8H)-one(B), tert-butyl4-(6-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7, 8-dihydropyrido ［2, 3-d］ pyrimidin-2-yl)amino)pyridin-3-yl)piperazi-ne-1-carboxylate(C). Based on the structures of the impurities, the possible routes to them were discussed, and their structures were elucidated by 1H NMR and MS.

Synthesis and biological evaluation of novel pleuromutilin derivatives with nitrogen-containing heterocycles as antibacterial agents / 药学学报

A series of new pleuromutilins derivatives were designed and synthesized through coupling 2-aminothiazole ring of WL001 with different nitrogen-containing substituted heterocycles in the side chain. Their biological activities were evaluated against both Gram-positive and Gram-negative clinical bacteria in vitro Most new compounds displayed specificity to certain strain of bacteria. Particularly, compounds with saturated nitrogen-containing heterocycles exhibited significant antibacterial activities (0.062 5-8 µg · mL(-1)) superior or similar to those of amoxicillin, tiamulin and levofloxcin. Furthermore, treatment with 15a and 15b having piperidine or morpholine residues also could effectively inhibit Gram-negative bacteria. Therefore, our novel findings may provide a new insight into the design of novel pleuromutilin derivatives and lay the basis for further studies on the treatment of drug-resistance of pathogenic bacteria.

Gene Cloning, Expression and Immunogenicity of the Protective Antigen Subolesin in Dermacentor silvarum

Subolesin (4D8), the ortholog of insect akirins, is a highly conserved protective antigen and thus has the potential for development of a broad-spectrum vaccine against ticks and mosquitoes. To date, no protective antigens have been characterized nor tested as candidate vaccines against Dermacentor silvarum bites and transmission of associated pathogens. In this study, we cloned the open reading frame (ORF) of D. silvarum 4D8 cDNA (Ds4D8), which consisted of 498 bp encoding 165 amino acid residues. The results of sequence alignments and phylogenetic analysis demonstrated that D. silvarum 4D8 (Ds4D8) is highly conserved showing more than 81% identity of amino acid sequences with those of other hard ticks. Additionally, Ds4D8 containing restriction sites was ligated into the pET-32(a+) expression vector and the recombinant plasmid was transformed into Escherichia coli rosetta. The recombinant Ds4D8 (rDs4D8) was induced by isopropyl beta-D-thiogalactopyranoside (IPTG) and purified using Ni affinity chromatography. The SDS-PAGE results showed that the molecular weight of rDs4D8 was 40 kDa, which was consistent with the expected molecular mass considering 22 kDa histidine-tagged thioredoxin (TRX) protein from the expression vector. Western blot results showed that rabbit anti-D. silvarum serum recognized the expressed rDs4D8, suggesting an immune response against rDs4D8. These results provided the basis for developing a candidate vaccine against D. silvarum ticks and transmission of associated pathogens.

The effect of NGF gene modified Scwhann cells on DRG neurons repair after compressed injury / 中华骨科杂志

ObjectiveTo study the effect and mechanism of NGF gene modified SCs on DRG (dorsal root ganglion) neurons repair after compressed injury.MethodsSCs were obtained by one enzyme digestion method.SCs were transfected with NGF gene by adenovirus.Thirty-two female SD rats with compression injury of dorsal root ganglia on right lumbar nerve roots.were divided randomly into following groups:Normal saline(NS) group,Pure SCs group,Ad-NGF group,and SCs+NGF group.Nerve root tissues were harvested 2 weeks after treatment.Western blot were used to detect the proNGF volume in nerve root tissue lysis; Double-labeling fluorescent Immunohistochemistry(IHC) was used to count the number of β-Tubulin Ⅲ positive cells and activating transcription factor 3 positive cells.The ratio of injured neurons to survived neurons was calculated.ResultsWestern bolt showed the proNGF volume in nerve root tissue lysis of SCs+NGF group increased dramatically.Double-labeling fluorescent IHC showed SCs+NGF group vs any group,the density of survived DRG neurons(β-Tubulin Ⅲ positive cells) increased significantly,meanwhile the percentage of injured neurons (ATF3 positive cells) in survived neurons decreased dramatically.Conclusion NGF gene Modified SCs could promote the survival of DRG neurons after compression injury and decrease the ratio of injured neurons.We conclude that this study provides a new treatment strategy for the patients who suffer from chronic compression injury on nerve roots and DRG neurons.

Cardiomyocyte Differentiation from P19 Cells Induced by NKX2-5 / 中国医科大学学报

Objective To identify the rale of NKX2-5 gene in cardiomyocyte differentiation and its mechanism.Methods P19 cells were divided into transfected and non-transfected groups.In the transfected group,P19 cells were with stable expression of NKX2-5 gene.The P19 cells were cultured in suspension for 4 days,and the formed aggregates were transferred to Petri dish for adherent culture.On days 4,8,12,and 16 of the adherent culture,the expressions of ct-saicomeric actin(α-SA)and cardiac troponin T(cTnT)were detected with double-labeling immunofluorescence and Western blot.The ultrastruetural changes were observed on day 16.Results In the transfected group,no expression of α-SA and cTnT was found on day 4,and the expression of these 2 proteins or co-expression existed on days 8,12,and 16.There were early cell junction and myofilament-like structure in the cytoplasm of some cells in the transfected group.In the non-transfected group,these 2 proteins were negative,and no differentiated cell was found.Conclusion Stable expression of NKX2-5 gene can induce cardiomyocyte differentiation from P19 cells,but the P19 cells with stable expression of JVKX2-5 gene is not suitable to be an in vitro model of cardiac development.