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@#Abstract: Objective To construct SARS-CoV-2 receptor binding domain molecular probe for monoclonal memory B cell sorting and obtain RBD specific neutralizing antibodies from peripheral blood mononuclear cells (PBMCs) of COVID-19 convalescents by single-cell sorting. Methods The SARS-CoV-2 RBD sequence was downloaded from GenBank, and the Avi tag and 6-histidine tags were added at the C-terminal. After codon optimization, it was chemically synthesized, cloned into the pDRVI1.0 vector, expressed after transfection of 293F cells, and biotinylated consequently. RBD-specific B cells were sorted out with this probe1 from the PBMCs of convalescents recovered from COVID-19. After B cells were lysed, the variable regions of heavy chain and light chain were amplified, cloned into the antibody expression vector, and transfected into 293F cells to express the antibody. Then the antibody was purified from the supernatant using protein A column and SARS-CoV-2 pseudovirus was used to test their neutralizing activity. Results RBD-Avi probe was produced and successfully biotinylated sequentially with an efficiency of 30%-50%. Western blot analysis revealed that the biotinylated probe was recognized by the antibodies purified from COVID-19 convalescent plasma. Using this probe, 7 and 16 RBD-specific memory B cells were successfully isolated from the PBMCs of two convalescent individuals, accounting for 0.24% and 0.17% of the total cell population, respectively. After amplifying the variable regions of antibody heavy and light chains from the lysed B cells, 7 and 12 pairs of antibody heavy-light chains were obtained. A total of 16 antibodies were expressed in the convalescent individuals, and most of the purified antibodies showed neutralizing activity against the pseudovirus, with IC50 values of 6 antibodies below 1 μg/mL. The IC50 values of XJ-A9 and SCF-F1 against the wild-type pseudovirus were 0.07 μg/mL and 0.35 μg/mL, respectively. Conclusion The SARS-CoV-2 RBD molecular probe constructed in this study has good antigenicity, and the isolated antibodies present neutralizing activity against wild-type SARS-CoV-2 pseudovirus.
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@#Abstract：Objective To carry out serological analysis of varicella⁃zoster virus（VZV）IgG antibody level in healthy people aged 1 ~ 30 years in Liaoning Province. Methods In October 2020，3～5 mL venous blood samples were collected from 617 healthy people aged 1～30 years selected from six counties and districts in Shenyang，Fuxin and Dandong of Liaoning Province by stratified random sampling method，of which serum samples were collected and determined for VZV IgG antibody level by ELISA. The positive rate of serum antibody and geometric mean concentration（GMC）of antibody were calculated and compared. Results Among 617 serum samples，302 samples were positive for VZV IgG antibody，the positive rate was 48. 947%，and the GMC was 112. 772 mIU／mL. The positive rate of VZV IgG antibody was 29. 670%～75. 789% and the GMC was 45. 508～366. 559 mIU／mL in healthy people of various ages. Both of the antibody positive rate（χ2 = 67. 104， P < 0. 001）and GMC（F = 20. 685，P < 0. 001）showed significant differences. The positive rates of VZV IgG antibody in male and female were 44. 817% and 53. 633% respectively，which showed significant difference（χ2 = 4. 779，P = 0. 029）， while the GMCs were 96. 983 and 133. 829 mIU／mL respectively（t = -1. 958，P = 0. 051）with no significant difference. The positive rates of VZV IgG antibody of healthy people in Shenyang，Fuxin and Dandong of Liaoning Province were 55. 224%，40. 201% and 51. 152% respectively with significant differences（χ2 = 9. 683，P = 0. 008），of which the positive rate of FuxinwassignificantlylowerthanthoseofShenyangandDandong（χ2 =9. 046and5. 013，P =0. 003and0. 025，respectively）； While the GMCs were 133. 523，85. 953 and 123. 713 mIU／mL respectively with no significant difference（F = 0. 514， P = 0. 598）. Among 617 serum samples，54 sampleswere suspicious，which remained within the criticalrange afterre⁃examina⁃ tion，while the gap between positive rate and the total percentage of positive and suspicious results gradually decreased with the increase of age，indicating that the immunity to varicella gradually increased with the increase of age. Conclusion The VZV⁃IgG antibody level of healthy people aged 1～30 years in Liaoning Province increased gradually with age，while the overall level was low. To control the spread of varicella virus，it is recommended to increase varicella vaccine coverage in vulnerable areas and susceptible population to build VZV immune barrier.
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OBJECTIVE@#To compare the efficacy of eltrombopag combined with cyclosporine A (CsA) and CsA alone in patients with transfusion-dependent non-severe aplastic anemia (TD-NSAA).@*METHODS@#The clinical data of 76 patients with treatment-naive TD-NSAA in Ningde Municipal Hospital of Ningde Normal University and Affiliated Hospital of Nantong University from December 2017 to June 2021 were retrospectively analyzed. Among them, 45 cases were treated with eltrombopag combined with CsA, and 31 patients with compatible baseline characters were treated with CsA alone. The efficacy of patients between the two groups was compared, and the factors affecting the curative effects were also analyzed.@*RESULTS@#There were significant differences in hematological response (HR) and complete response(CR) rates between the two groups at 3, 6, 12 months, and follow-up endpoint of treatment (P<0.05). With the prolongation of eltrombopag treatment time, the curative effect increased gradually, and the patients achieved more CR and HR rates by the end of the follow-up period. Simultaneously, with the increase in the maximum stable dose of eltrombopag, the HR rate increased gradually. The megakaryocyte count in eltrombopag group was higher than that in control at 6 and 12 months (P<0.05). Compared with the control group, the median time of platelet transfusion independence in eltrombopag group was more shorter (P=0.018), and the median platelets transfusion volume was lower (P=0.009). At 3, 6, 12 months after eltrombopag, the change of platelet in eltrombopag group was higher than that in the control group (P<0.05). Analysis of related factors affecting the efficacy showed that sex, age, iron overload, platelet count before treatment had no effect on the efficacy, and the median maximum stable dosage and the administration period for eltrombopag were related to the curative effect. The patients of eltrombopag group experienced adverse events of varying degrees, but the reactions were mild and mostly tolerated.@*CONCLUSION@#Eltrombopag can effectively improve the hematopoietic response and promote platelet recovery for TD-NSAA patients with relatively more residual hematopoietic cells, and it is safe and well tolerated.
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Humans , Anemia, Aplastic/therapy , Retrospective Studies , Treatment Outcome , Cyclosporine/therapeutic use , Immunosuppression Therapy , Immunosuppressive Agents/therapeutic useABSTRACT
@#Abstract: AIDS combined with Pneumocystis jirovecii pneumonia (PJP) and disseminated infections of Talaromyces marneffei and Cryptococcus neoformans are rare. This paper summarizes and analyzes the diagnosis and treatment of an AIDS patient with multiple fungal infections for reference. A 79-year-old male patient was admitted to the hospital with "stool habit change for more than 20 days". The white blood cell count was 4.57×109/L, the percentage of neutrophils was 81.8%, the absolute count of CD4+ lymphocytes was 6/μL, and the CD4/CD8 ratio was 0.17. HIV antibody positive was confirmed by CDC. The cerebrospinal fluid and alveolar lavage fluid were positive for Cryptococcus neoformans capsular antigen, and Pneumocystis jirovecii was found by the bronchoalveolar lavage fluid stained with hexamine silver. The cerebrospinal fluid culture was positive for Cryptococcus neoformans, and the blood culture was positive for Cryptococcus neoformans and Talaromyces marneffei. CT showed that bronchovascular bundles in both lungs were more thick, patchy and cable-like high-density shadows were seen in both lungs, and the edges were blurred. Nodular and cable-like high-density shadows were seen in the posterior apical segment of the left upper lobe, with clear margins. Infection of both lungs was considered, and secondary pulmonary tuberculosis occurred in the left upper lobe. After admission, the patient was treated with various anti-bacterial and fungal drugs due to recurrent fever, but the effect was not effective. The fever symptoms of the patient could not be significantly improved, and his condition continued to worsen, and he eventually died. The patient with AIDS complicated with bacterial and fungal infection, especially PJP infection in serious condifiton and has a poor prognosis for rapid development, so clinical attention should be paid to.
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Objective To investigate the effectiveness and cost of 50% and 80% wettable powder of niclosamide ethanolamine salt (NESWP) and 26% metaldehyde and niclosamide suspension concentrate (MNSC) in hilly schistosomiasis-endemic regions, so as to provide insights into the selection of chemical molluscicides in hilly regions. Methods In September 2020, a wasteland in Guanshanqiao Village, Yanrui Township, Yushan County of Jiangxi Province was selected as the experimental region, which was sectioned into five blocks and defined as four experimental groups (A1, A2, B, C) and a blank control group (D). 80% NESWP were given at doses of 1 g/m2 and 1.5 g/m2 in groups A1 and A2 using the spraying method, 50% NESWP was given at a dose of 2 g/m2 in Group B using the spraying method, and 26% MNSC was at a dose of 4 g/m2 in Group C using the spraying method, while no chemical treatment was given in Group D. Snail survey was performed using a systematic sampling method before chemical treatment and 1, 3, 7 d and 15 d post-treatment to examine the molluscicidal effect, and all molluscicidal costs were estimated to calculate the cost of chemical treatment per 1 m2 and the cost of the reduction in the mean density of living snails per 1%. Results The highest mortality of snails was 78.95% and the lowest density of living snails was 0.2388 snails/0.1 m2 in the experimental groups within 7 d of chemical treatment, and the highest mortality of snails was 94.74% and the lowest density of living snails was 0.058 0 snails/0.1 m2 7 d post-treatment. There were no significant differences in the snail mortality among the A1, A2, B and C groups 1 (χ2 = 2.250, P > 0.05), 3 (χ2 = 1.779, P > 0.05) or 15 d post-treatment (χ2 = 2.286, P > 0.05), while a significant difference was detected in the snail mortality among the four groups 7 d post-treatment (χ2 = 7.990, P = 0.046). In addition, there were no significant differences in the snail mortality between A1 and A2 groups 1 (χ2 = 0.724, P > 0.05), 3 (χ2 = 0.584, P > 0.05), 7 (χ2 = 0.400, P > 0.05) or 15 d post-treatment (χ2 = 0.251, P > 0.05). The costs of chemical treatment per 1 m2 were 0.58, 0.60, 0.64 Yuan and 0.73 Yuan in groups A1, A2, B and C, and the costs of the mean density of living snail per 1% reduction were 19.29, 20.44, 21.68 Yuan and 23.53 Yuan in groups A1, A2, B and C, respectively. Conclusion 80% NESWP shows a high molluscicidal efficacy and low cost in hilly schistosomiasis-endemic regions.
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Objective: To investigate the effect and mechanism of cyclanoline on nitrosamine (BBN)-induced bladder cancer in rats. Methods: Male SD rats were randomly divided into control group, model group, Cyc-low dose (20 mg/kg) group, Cyc-high dose (40 mg/kg) group and cisplatin (5 mg/kg) group. Rats were intragastrically administered with 0.5 mL BBN (0.4 kg/L, twice a week for 8 weeks) to establish bladder cancer model. In addition to control and model group given with normal saline, the other groups were intraperitoneally injected with drugs (once a day for 8 weeks). The bladder tissue was collected after experiment. HE staining was used to investigate the histopathological changes of the bladder in rats. The expressions of MMP9 and Ki67 were observed by immunohistochemistry method, and the expressions of KLF4, p21, CyclinD1, E-cadherin, N-cadherin, Vimentin, Wnt, β-catenin in the bladder tissues were detected by Western bloting. Results: Compared with the model group, cyclanoline effectively inhibited the infiltration of cystitis cells, promoted the degeneration of cancer cells, and reduced the proportion of cytoplasm. Cyclanoline significantly decreased the expressions of MMP9 and Ki67 (P < 0.05, 0.01), up-regulated the expressions of KLF4, p21 and E-cadherin (P < 0.05, 0.01), down-regulated the expressions of CyclinD1, Wnt, β-catenin, N-cadherin and Vimentin in bladder cancer tissues (P < 0.05, 0.01). Conclusion: Cyclanoline promotes the expression of KLF4, inhibits Wnt/β-catenin signaling transduction and the epithelial cell-mesenchymal transition of bladder cancer cells, thereby inhibiting the migration and invasion of bladder cancer cells. Meanwhile, cyclanoline regulates the expressions of p21 and CyclinD1 by up-regulating KLF4, affects the proliferation of bladder cancer cells, and thereby delays the pathological process of BBN-induced bladder cancer in rats.
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Objeetive@#To learn the sleep behaviors of preschool children and their association with behavioral problems,so as to provide reference for improving sleep quality and behaviors in preschool children.@*Methods@#By stratified random sampling method,the children aged 3-6 years were selected from 11 counties in 5 cities of Zhejiang Province. A self-designed questionnaire was used to investigate 19 items of sleep behaviors, and Achenbach children's behavior Scale（CBCL）was used to evaluate behavior problems. Spearman rank correlation was conducted to analyze the relationship between sleep behaviors and behavioral problems in preschool children. @*Results @#Totally 1 865 preschool children were enrolled,1 732 valid questionnaires were collected,with a response rate of 92.86%. The total score of sleep behaviors in preschool children was 5.87±3.38. The prevalence rates of 19 sleep behaviors ranged from 0.69% to 82.85%;the common behaviors were rejecting quilt cover（82.85%）,grinding teeth（53.35%）,sleep-talking（53.23%）,drooling（52.66%）and snoring（43.07%）. The total score of CBCL was 20.01±14.39. There were 229 children with behavioral problems,accounting for 13.22%. The results of Spearman rank correlation showed that except for sleeping posture and apnea,all the other 17 items of sleep behaviors were correlated with the total score of CBCL（P < 0.05）. @*Conclusion@#Preschool children generally have problems in sleep, their sleep behaviors are correlated with behavioral problems.
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@#[Abstract] Objective: To explore the clinical significance and in vitro biological effect of ubiquitin carboxyl-terminal hydrolase L5 (UCHL5) expression in thyroid carcinoma (TC) tissues. Methods: TCGAdata were used to analyze the expression of UCHL5 in thyroid carcinoma tissues and its relationship with the prognosis of patients. 82 pairs of TC tissues and corresponding adjacent tissues were collected in the Department of Vascular and Thyroid Surgery, Sichuan Provincial People's Hospital from May 2018 to July 2019; TC cell lines (KTC-1 and WRO) were cultured in vitro, and transfected with UCHL5 overexpression vectors or their control vectors via lentivirus. The mRNAand protein expressions of UCHL5 and B-Raf proto-oncogene serine/threonine-protein kinase (BRAF) in tissues and cells were detected by qPCR and Western blotting, respectively. Cell proliferation was detected by CCK-8, and cell invasion and migration were detected by Transwell and Wound-healing experiments. Results: The expression of UCHL5 was low in TC tissues (P<0.01), and its expression was upregulated in tumor tissues with high TNM stage (P<0.01). The expression of UCHL5 was significantly correlated with BRAF expression and TNM stage of patients (all P<0.01), but not significantly related with patient's age, gender, pathological type and BRAF mutation (all P>0.05). In vitro overexpression of UCHL5 in KTC-1 and WRO cells could significantly promote BRAF expression, cell proliferation and metastasis (all P<0.01). Conclusion: The expression of UCHL5 is low in TC tissue, but upregulated with tumor progression. The high expression of UCHL5 in TC patients suggests poor prognosis. Meanwhile, UCHL5 can promote the malignant behaviors of TC cells in vitro.
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Nineteen preschisanartane-type schinortriterpenoids (SNTs), among which eleven ones were previously undescribed, were isolated from two Schisandra species, S. sphaerandra and S. rubriflora. Their structures were determined using 1D and 2D NMR spectroscopic analyses, NMR data comparison, quantum chemical calculation of NMR parameters, electronic circular dichroism (ECD), X-ray single crystal diffraction, and chemical derivation. Furthermore, structural re-examination of a few previously reported preschisanartane-type SNTs led to the structural revision of preschisanartanin J. Besides, it is suggested that the reported structures of arisanlactone D and schilancidilactone W should be re-checked. Finally, a few isolated SNTs were found to possess neurite outgrowth-promoting activities, and protective activities against neural injuries.
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Wu-tou decoction (WTD) was originally recorded in the synopsis of the golden chamber and it had been widely used for the treatment of neuropathic pain (NP) with exact therapeutic efficacy. However, the underlying molecular mechanisms still remain unclarified. Thus, in this research, we aimed at clarifying the underlying molecular mechanisms of WTD against NP by combining network analysis and experimental validation based on the spinal nerve ligation (SNL) model. Firstly, the network analysis indicated that key targets of WTD were significantly involved in the MAPK signaling pathway (P = 4.04E-12) and four important components of the above pathway, AKT kinase (AKT), MAP kinase kinase 4 (MKK4), c-Jun N-terminal kinase (JNK) and transcription factor AP-1 (JUN) had been reported to play a vital role in neuroinflammation during the disease process of NP. Then, experimental validation results proved that WTD markedly reduce the severity of mechanical allodynia (P<0.01) and cold hypersensitivity (P<0.05) of SNL rats. In addition, Western blot results provided evidence that the phosphorylated protein expression levels of AKT, MKK4, JNK and JUN in the superficial lamina of spinal cord of SNL rats were markedly increased (P<0.001), and WTD could improve the phosphorylated protein expression level of AKT (P<0.001) which was reported to be nerve protective and attenuate the phosphorylated protein expression levels of MKK4, JNK and JUN (P<0.01) which were closely involved into neuroinflammation. In conclusion, this study indicated that WTD might exert anti-hyperalgesia action through the inhibition of neuroinflammation mediated by AKT-MKK4-JNK-JUN which belong to the MAPK signaling pathway. These findings also provided scientific evidences that WTD might be a promising candidate for NP. Animal experiments in this study were approved by the Ethics Committee of Experimental Animals of the China Academy of Chinese Medical Sciences.
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In 1980s, Biomphalaria straminea, an intermediate host of Schistosoma mansoni, was found in Shenzhen City, Guangdong Province, China, and currently, this snail has colonized in Shenzhen City and spread to peripheral cities involving of Dongguan and Huizhou. Since imported cases infected with S. mamoni have been reported from time to time in China, Mainland China is facing the potential risk of transmission of schistosomiasis mansoni. With the deepening of the opening-up policy, notably the implementation of the Belt and Road Initiative, there is an increase in the risk of transmission of schistosomiasis mansoni in Mainland China. Increasing the understanding on schistosomiasis mansoni, improving the awareness toward schistosomiasis mansoni prevention and control, and identifying, reporting and managing imported cases with S. mansoni infection or pathogen carriers, are of particular importance to prevent the development of entire life cycle of S. mansoni and the resultant schistosomiasis mansoni transmission in China. To protect public health, a consensus has been reached pertaining to the surveillance and control strategy of imported schistosomiasis mansoni by Chinese infectious disease experts and parasitologists, with aims to improve the awareness and capability for the diagnosis, treatment and control of imported schistosomiasis mansoni among Chinese disease control and prevention institutions and medical institutions, and decrease and even eliminate the risk of schistosomiasis mansoni transmission in China.
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In 1980s, Biomphalaria straminea, an intermediate host of Schistosoma mansoni, was found in Shenzhen City, Guangdong Province, China, and currently, this snail has colonized in Shenzhen City and spread to peripheral cities involving of Dongguan and Huizhou. Since imported cases infected with S. mamoni have been reported from time to time in China, Mainland China is facing the potential risk of transmission of schistosomiasis mansoni. With the deepening of the opening-up policy, notably the implementation of the Belt and Road Initiative, there is an increase in the risk of transmission of schistosomiasis mansoni in Mainland China. Increasing the understanding on schistosomiasis mansoni, improving the awareness toward schistosomiasis mansoni prevention and control, and identifying, reporting and managing imported cases with S. mansoni infection or pathogen carriers, are of particular importance to prevent the development of entire life cycle of S. mansoni and the resultant schistosomiasis mansoni transmission in China. To protect public health, a consensus has been reached pertaining to the surveillance and control strategy of imported schistosomiasis mansoni by Chinese infectious disease experts and parasitologists, with aims to improve the awareness and capability for the diagnosis, treatment and control of imported schistosomiasis mansoni among Chinese disease control and prevention institutions and medical institutions, and decrease and even eliminate the risk of schistosomiasis mansoni transmission in China.
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Objective The aim of this study was to investigate the effect of Ca2+ /calmodulin - dependent kinase II (CaMKII)γ RNA interference on the expression of nuclear factor of activated T-cells cytoplasmic 1(NFATc1),tyrosine kinase(c-Src)and tartrate resistant acid phosphatase(TRAP)genes,and its role and molecular mechanism in osteoclast differentiation. Methods The CaMKII γ RNA interference vector was constructed by lentivirus and transfected into RAW264. 7 cells. The experiment was di-vided into three groups:A,B and C,which were the control group,negative vector group and interference vector group. After transfec-tion for 12 hours,osteoclasts induced by 50 ng/mL RANKL and the cells were harvested after induction for 5 days. Real-time quanti-tative PCR,Western blot and immunofluorescence were used to detect the expression of NFATc1,TRAP and c-Src genes in three groups. Results The mRNA levels of NFATc1,TRAP and c-Src in the group C decreased by 49. 86% ,43. 65% and 53. 57% ,re-spectively(P<0. 001),and the protein levels decreased by 54. 22% ,46. 75% and 45. 86% ,respectively(P<0. 001). There was no significant difference between the A and the B groups(P>0. 05). The fluorescence intensity of the above genes in the group C was significantly weaker than that in the A and B groups,and the formation of osteoclasts was significantly less than that in the A and B groups. Conclusion CaMKIIγ RNA interference significantly inhibited the expression of NFATc1,TRAP and c-Src genes,sugges-ting that CaMKIIγ plays a key regulatory role in osteoclast differentiation.
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OBJECTIVE@#To evaluate the clinical efficacy of low dose combined chemotherapy（LDCC） for patients with relapsed and refractory aplastic anemia-paroxysmal nocturnal hemoglobinuria（AA-PNH） syndrome, and to analyze the advantages of LDCC in the treatment of AA-PNH syndrome.@*METHODS@#The clinical characteristics and the curative effect of LDCC in 9 patients with relapsed and refractory AA-PNH syndrome were retrospectively analyzed. Five patients were treated with MP therapy［melphalan 2 mg/(m·d); prednisone 0.5 mg/(kg·d)］, and the other 4 patients were treated with HA therapy(HHT 2 mg/d iv drip, for 5 days; Ara-C 100 mg/d iv drip, for 5 days). The changes of PNH clone, dosage of corticosteroid, hemolysis and the relapse of disease, hematological parameters and adverse reactions were compared before and after therapy. All patients were treated for 1-2 courses.@*RESULTS@#Seven out of 9 patients responded well, the dosage of corticosteroid and the bilirubin concentration decreased significantly and anemia was relieved in 7 patients (P<0.05). One patient relapsed in one year. PNH clone of 3 patients turned negative. Five patients did not rely on blood transfusion in 1 year. There was no bone marrow failure to be found in all patients.@*CONCLUSION@#The LDCC has better efficacy and safety in the treatment of patients with AA-PNH syndrome, moreover, the patients is more tolerant to LDCC, thus the LDCC may be a selection for treatment of patients with relapsed and refractory AA-PNH syndrome.
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Humans , Anemia, Aplastic , Anemia, Refractory , Hemoglobinuria, Paroxysmal , Hemolysis , Retrospective StudiesABSTRACT
To study the SSR loci information and develop molecular markers, a total of 43 683 Unigenes in transcriptome of Andrographis paniculata were used to explore SSR. The distribution frequency of SSR and the basic characteristics of repeat motifs were analyzed using MicroSAtellite software, SSR primers were designed by Primer 3.0 software and then validated by PCR. Moreover, the gene function analysis of SSR Unigene was obtained by Blast. The results showed that 14 135 SSR loci were found in the transcriptome of A. paniculata, which distributed in 9 973 Unigenes with a distribution frequency of 32.36%. Di-nucleotide and Tri-nucleotide repeat were the main types, accounted for 75.54% of all SSRs. The repeat motifs of AT/AT and CCG/CGG were the predominant repeat types of Di-nucleotide and Tri-nucleotide, respectively. A total of 4 740 pairs of SSR primers with the potential to produce polymorphism were designed for maker development. Ten pairs of primers in 20 pairs of randomly picked primers produced fragments with expected molecular size. The gene function of Unigenes containing SSR were mostly related to the basic metabolism function of A. paniculata. The SSR markers in transcriptome of A. paniculata show rich type, strong specificity and high potential of polymorphism, which will benefit the candidate gene mining and marker-assisted breeding.
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Objective To investigate the function of cAMP response element binding protein (CREB) and extracellular signal-regulated kinase (ERK1/2) in osteoclast differentiation mediated by Ca2+/calmodulin-dependent kinase Ⅱ (CaMK Ⅱ)δ,and elucidate the molecular mechanism thereof.Methods CaMK Ⅱδ RNA interference lentivirus vector was constructed and mouse RAW264.7 cells were transfected with the virus to determine the interference efficiency.After virus transfection,RAW264.7 cells were treated with 50ng/ml receptor activator of nuclear factor κB ligand (RANKL) and the phosphorylation levels of CREB and ERK1/2 were detected at different time points.The cells were also treated with PD98059,an ERK1/2 inhibitor,to determine the effect of ERK1/2 signal blocking on the expression of nuclear factor activated T-cells cytoplasmic 1 (NFATc1) and osteoclast differentiation.Results Interference efficiency of recombinant CaMK Ⅱδ virus vector was 77.2％ at mRNA level and 70.2％ at protein level.CaMK Ⅱδ RNA interference significantly suppressed phosphorylation of CREB and ERK1/2,and the levels ofp-CREB and p-ERK1/2 were down-regulated by 21％-55％ and 55％-64％,respectively.ERK1/2 inhibitor significantly down-regulated the protein expression of NFATc1,and the number of osteoclast,the number and size of bone resorption lacunae decreased by 39.3％,50.0％ and 52.3％,respectively.Conclusion CaMK Ⅱδ RNA interference may significantly suppress the phosphorylation of CREB and ERK1/2,and CREB and ERK1/2 have mediated the CaMK Ⅱδ-induced osteoclast differentiation.
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Objective: To compare the effect of galectin-3 (Gal-3), NT-proBNP and echocardiography paramerters on assessing cardiac function in patients with chronic heart failure (HF). Methods: A total of 144 patients treated in our hospital from 2016-03 to 2016-11 were enrolled. According to the NYHA classification, the patients were divided into 2 groups: HF group and Normal cardiac function group. n=72 in each group. Basic clinical information was collected, blood levels of Gal-3 and NT-proBNP were examined, echocardiography was conducted to measure left ventricular ejection fraction (LVEF) and left ventricular end diastolic diameter (LVEDD). Correlations between Gal-3, NT-proBNP and echocardiography parameters were studied, the abilities of Gal-3, NT-proBNP and echocardiography for estimating HF were compared. Results: Compared with Normal cardiac function group, HF group had increased blood levels of NT-proBNP [3499.5 (1431.3-9088.0) ng/L] vs [384.1 (122.1-1540.5) ng/L] and Gal-3 [3.0 (1.71-5.8) pg/ml] vs [1.9 (1.4-2.6) pg/ml], decreased LVEF [49.5% (42%-58%)] vs [62.5% (59%-67%)], enlarged LVEDD [52.0 (46.3-57.8) mm] vs [46.0 (42.0-49.0) mm] and elevated serum creatinine [113.6 (90.5-152.7) umol/L] vs 82.4 (69.1-97.4) umol/L], all P<0.05. Correlation analysis showed that NT-proBNP and Galectin-3 were positively related to LVEF and LVEDD; Gal-3 and NT-ProBNP had the strongest correlation (r=0.57, P<0.01). The AUC of ROC for Gal-3 was 0.674 (0.584-0.763), for NT-proBNP was 0.837 (0.771-0.902) and for LVEF was 0.806, (0.735-0.878) which implied that NT-proBNP was the most powerful parameter for estimating HF. Conclusion: Gal-3 had the ability to estimate HF and could be used as a biomarker, while its ability was lower than NT-proBNP in clinical practice.
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Objective :To study correlation between treadmill exercise test assessing myocardial ischemia and coronary stenosis .Methods :A total of 150 patients with stable angina pectoris in our hospital ,according to vascular stenosis by coronary CT were divided into group A (stenosis＜ 50%,n＝ 50) ,group B (stenosis 50% ～75%,n＝ 50) and group C (stenosis ＞75%,n＝50).Coronary stenotic rate and ST depression during exercise test were compared a-mong three groups.Spearman method was used to analyze correlation between treadmill exercise positive rate and coronary stenosis .Results :Compared with group B ,there were significant rise in coronary diameter stenotic rate [(63.64 ± 4.21)% vs.(66.71 ± 4.46)%] and vascular diameter of reference segment [(2.92 ± 0.23) mm vs.(3.03 ± 0.21) mm] in group C ,P＜0.05 or ＜0.01. Along with coronary stenosis aggravated ,there was significant re-duction in onset time of ST depression [(712.3 ± 202.7) s vs.(602.3 ± 210.4) s vs.(501.2 ± 236.1) s] ,and signif-icant rise in duration [ (425.4 ± 200.5) s vs.(520.8 ± 205.8) s vs.(603.4 ± 198.4) s] and treadmill exercise posi-tive rate (64.00% vs.82.00% vs .92.00%) ,in group A ,B ,C respectively ;and ST depression extent of group C was significantly higher than that of group A [(2.4 ± 1.1) mV vs.(1.9 ± 0.8) mV] ,P＜0.05 or ＜0.01. Spearman correlation analysis indicated that treadmill exercise positive rate (myocardial ischemia rate ) was not significantly correlated with coronary stenotic degree (r＝3.425 , P＝0.126).Conclusion :Although treadmill exercise test pos-sesses important significance in diagnosis of myocardial ischemia ,but coronary stenotic degree is not correlated with myocardial ischemic degree .which should raise clinical attention
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Objective To analyze the effect of Fufang Xueshuantong capsule on glaucomatous optic atrophy and its influence on ocular hemodynamics.Methods 124 cases of patients with glaucomatous optic atrophy from April 2014 to December 2016 in our hospital were selected,and randomly divided into experimental group and control group according to the order of hospital admission , with 62 cases in each group.The control group was treated with nerve growth factor plus vitamin needling,the experimental group was treated with nerve growth factor and vitamin and compound Xueshuantong capsule.Observation and comparison the total effective rate, visual function (visual acuity, total gray value) improvement of the two groups of patients, as well as the changes of ocular hemodynamic parameters (systolic peak velocity, diastolic End-stage blood flow velocity, resistance index, pulsatility index).Results The total effective rate of the experimental group was 93.55%(58/62), the total effective rate of the control group was 70.97%(44/62), the total effective rate of the experimental group was higher than the control group (P <0.05)).After treatment, the visual acuity of the experimental group was significantly higher than the control group, the total gray value of the experimental group was significantly lower than the control group (P<0.05).The peak systolic velocity and end diastolic velocity in the experimental group were significantly higher than those in the control group (P<0.05).The resistance index and pulsatility index in the experimental group were lower than those in the control group (P<0.05). Conclusion Compound Xueshuantong capsule is effective in the treatment of glaucomatous optic atrophy , can improve the ocular hemodynamics and improve the visual function.
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Objective To study the effect of zoledronate (ZOL) on Ca2+/calmodulin-dependent kinase Ⅱ δ (CaMK Ⅱ δ) and down-stream gene expressions during osteoclast differentiation.Methods Mouse osteoclast precursors RAW264.7 cells were divided into the control group and ZOL group.The cells in both groups were induced with 50μg/L receptor activator of nuclear factor kappa B ligand (RANKL) and were harvested on 5 d,while the cells in ZOL group were also simultaneously treated with 1 × 10-6 mol/L ZOL for 2 d.Five days later,the cells were harvested and examined osteoclastogenesis,as well as gene expressions of CaMK Ⅱ δ,nuclear factor of activated T-cells cytoplasmic 1 (NFATc1),tartrate-resistant acid phosphatase (TRAP) and cell-sarcoma receptor coactivator (c-Src).Results The number of TRAP positive multinuclear osteoclasts,number and size of dentin absorption lacunae and area in the ZOL group were (20.0±3.2),(18.0±4.2) and (6 335.3± 1 043.2)μm2 respectively,which were significantly lower than (36.0 ± 8.4),(37.2 ± 5.0) and (11 636.2 ± 3 661.1) μm2 in the control group and decreased by 44.4 ％,51.6 ％ and 45.6 ％ respectively (P＜0.01).ZOL also significantly inhibited the gene expressions of CaMK Ⅱ δ,NFATc1,TRAP and c-Src,and the mRNA levels of these genes were decreased by 44.1％,49.0％,53.8％ and 49.6％ respectively,the protein level were decreased by 43.5 ％,32.2 ％,45.5 ％ and 48.0 ％ respectively.The immunofluorescent cytochemistry detection results showed the fluorescence intensity of CaMK Ⅱ δ,NFATc1,TRAP and c-Srcin in the ZOL group was significantly weakened when compared with the control group.Conclusion ZOL could significantly inhibit the osteoclast formation and bone absorption function,and down-regulates gene expressions of CaMK Ⅱ δ,NFATc1,TRAP and c-Src in osteoclast differentiation.