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1.
Orthop Surg ; 14(5): 946-954, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35441488

RESUMO

OBJECTIVE: To observe expression of CD38, a key modulator of nicotinamide dinucleotide (NAD+) metabolism in mice with knee osteoarthritis, and protective effect of CD38 inhibition during the osteoarthritis (OA) development. METHOD: The destabilization of the medial meniscus (DMM) model was performed in mice to mimic the process of OA. Immunofluorescence of CD38 was performed to evaluate its response during the OA process. Limb bud-derived mesenchymal cells were isolated for micromass culture. 100 nM or 1 µM CD38 inhibitor (78c) treatment for 14 days and CD38 sgRNA infection were then used to explore the effects of chondrogenic differentiation via Alcian blue staining. The expressions of chondrogenic markers were detected using RT-PCR and Western blot. To explore the protective effect of CD38 inhibitor on cartilage degradation during OA in vivo, a CD38 inhibitor was injected into the knee joint after DMM operations. Micro-CT analysis and Safranin O-fast green staining were used to evaluate subchondral bone micro-architecture changes and cartilage degeneration. RESULTS: Compared to the control group, the CD38 expression in superficial cartilage was obviously increased in DMM group (P < 0.05). During the normal chondrogenic differentiation, the extracellular matrix formed and expression of Sox9, Col2, aggrecan increased apparently while CD38 expression decreased, which could be reversed with ablation of CD38 in limb bud-derived mesenchymal cells. Consistent with findings in vitro, CD38 blockage via CD38 inhibitor injection protected against osteosclerosis in medial subchondral bone and cartilage degeneration in DMM-induced experimental mice. Compared to the Sham group, DMM mice showed significantly increased values of BV and BV/TV in subchondral bone (P < 0.05) and Mankin score, which could be rescued by 78c treatment (P < 0.05). Also the CD38 inhibitor contributed to homeostasis of anabolism and catabolism by upregulating Sox9, Col2, aggrecan and downregulating Runx2, Col10 and Mmp13. CONCLUSION: This study primarily implicates CD38 as an important regulator of chondrogenic differentiation. Inhibition of CD38 demonstrated protection against cartilage degeneration, which suggests that CD38 could be a potential therapeutic target for OA.


Assuntos
ADP-Ribosil Ciclase 1 , Cartilagem Articular , Glicoproteínas de Membrana , Osteoartrite do Joelho , ADP-Ribosil Ciclase 1/metabolismo , Agrecanas , Animais , Cartilagem Articular/fisiopatologia , Condrócitos , Modelos Animais de Doenças , Homeostase , Glicoproteínas de Membrana/metabolismo , Meniscos Tibiais/cirurgia , Camundongos , Osteoartrite do Joelho/metabolismo
2.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 53(1): 109-113, 2022 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-35048609

RESUMO

OBJECTIVE: To investigate the relationship between sleep duration and depressive symptoms in older people in China, and to explore whether there are gender differences in the relationship. METHODS: Accessing the data from China Health and Retirement Longitudinal Study (CHARLS) for 2015 and 2018, we covered in the study a total of 2898 respondents, including 1684 males (58.1%) and 1214 females (41.9%). The 10-item form of Center for Epidemiological Studies Depression Scale (CES-D) was used to measure the depressive symptoms of the older people. The effects of sleep duration on depressive symptoms of older people in China were analyzed with the logistic model. Based on the depressive and non-depressive subgroups in 2018, the differences of health factor variables and sleep duration between 2015 and 2018 were studied. Adjustments were made for health factors presenting statistical differences in 2018 to further explore the stability of the analysis results. RESULTS: Among the female older people, those with short sleep duration and those with long sleep duration had increased risks of developing depressive symptoms [odds ratio ( OR)=1.815, 95% confidence interval ( CI): 1.357-2.429 and OR=1.364, 95% CI: 1.001-1.854, respectively] in comparison to those with normal sleep duration. Among the male older people, no connection was identified between sleep duration and risks for depressive symptoms ( P>0.05). Residing in rural areas, suffering from chronic diseases and having limited ability for Instrumental Activities of Daily Living (IADL) were risk factors for developing depressive symptoms ( P<0.05). The sleep duration for 2015 and that for 2018 were found to be statistically different in the depressive older people ( P<0.001), but no no statistically significant difference was found between the sleep duration for 2015 and that for 2018 among the non-depressive older people. Findings for chronic diseases and IADL for 2015 and those for 2018 were statistically different in the depressive older population ( P<0.001). In the non-depressive older population, findings for chronic diseases, IADL and tobacco smoking status for 2015 and those for 2018 showed statistical difference ( P<0.05). Further sensitivity analysis showed that in the female older population, short sleep duration increased the risks for depressive symptoms ( OR=1.819, 95% CI: 1.356-2.440, P<0.001), while long sleep duration did not affect the risks for depressive symptoms. In the male older population, short or long sleep duration were not associated with the risks for depressive symptoms. CONCLUSION: The association between sleep duration and the risks for depressive symptoms was different for men and women. For the female older population, the possibility of depressive symptoms should be considered for those with short or long sleep duration. For the male older population, short or long sleep duration had little impact on the risks for depressive symptoms.


Assuntos
Atividades Cotidianas , Depressão , Idoso , China/epidemiologia , Estudos de Coortes , Depressão/epidemiologia , Depressão/etiologia , Feminino , Humanos , Estudos Longitudinais , Masculino , Sono
3.
Appl Microbiol Biotechnol ; 105(24): 9153-9166, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34837493

RESUMO

To directly produce ß-alanine from glucose by microbial fermentation, a recombinant Corynebacterium glutamicum strain with high efficiency of ß-alanine production was constructed in this study. To do this, the biosynthetic pathway of ß-alanine in an L-lysine-producing strain XQ-5 was modified by enhancing carbon flux in biosynthetic pathway and limiting carbon flux in competitive pathway. This study showed that replacement of L-aspartate kinase (AK) with wild-type AK and disruption of lactate dehydrogenase and alanine/valine aminotransferases increase ß-alanine production because of decreasing the by-products accumulation. Moreover, L-aspartate-α-decarboxylase (ADC) from Bacillus subtilis was designed as the best enzyme for increasing ß-alanine production, and its variant (BsADCE56S/I88M) showed the highest activity for catalyzing L-aspartate to generate ß-alanine. To further increase ß-alanine production, expression level of BsADCE56S/I88M was controlled by optimizing promoter and RBS, indicating that Pgro plus ThirRBS is the best combination for BsADCE56S/I88M expression and ß-alanine production. The resultant strain XQ-5.5 produced 30.7 ± 2.3 g/L of ß-alanine with a low accumulation of lactate (from 5.2 ± 0.14 to 0.2 ± 0.09 g/L) and L-alanine (from 7.6 ± 0.22 to 3.8 ± 0. 32 g/L) in shake-flask fermentation and produced 56.5 ± 3.2 g/L of ß-alanine with a productivity of 0.79 g/(L·h) and the glucose conversion efficiency (α) of 39.5% in feed-batch fermentation. This is the first report of genetically modifying the biosynthetic pathway of ß-alanine that improves the efficiency of ß-alanine production in an L-lysine-producing strain, and these results give us a new insight for constructing the other valuable biochemical. KEY POINTS: • Optimization and overexpression of the key enzyme BsADC increased the accumulation of ß-alanine. • The AK was replaced with wild-type AK to increase the conversion of aspartic acid to ß-alanine. • A 56.5-g/L ß-alanine production in fed-batch fermentation was achieved.


Assuntos
Corynebacterium glutamicum , Vias Biossintéticas/genética , Corynebacterium glutamicum/genética , Glucose , Engenharia Metabólica , beta-Alanina
4.
Zhongguo Zhong Yao Za Zhi ; 46(10): 2481-2488, 2021 May.
Artigo em Chinês | MEDLINE | ID: mdl-34047094

RESUMO

The paclitaxel-loaded and folic acid-modified poly(lactic-co-glycolic acid) nano-micelles(PTX@FA-PLGA-NMs) were prepared by the emulsion solvent evaporation method, and the parameters of paclitaxel-loaded nano-micelles were optimized with the particle size and PDI as evaluation indexes. The morphology of the nano-micelles was observed by transmission electron microscopy(TEM), and the stability, drug loading and encapsulation efficiency were systematically investigated. In vitro experiments were performed to study the cytotoxic effects of nano-micelles, apoptosis, and cellular uptake. Under the optimal parameters, the nano-micelles showed the particle size of(125.3±1.2) nm, the PDI of 0.086±0.026, the zeta potential of(-20.0±3.8) mV, the drug loading of 7.2%±0.75%, and the encapsulation efficiency of 50.7%±1.0%. The nano-micelles were in regular spherical shape as observed by TEM. The blank FA-PLGA-NMs exhibited almost no inhibitory effect on the proliferation and growth of tumor cells, while the drug-loaded nano-micelles and free PTX exhibited significant inhibitory effects. The IC_(50) of PTX@FA-PLGA-NMs and PTX was 0.56 µg·mL~(-1) and 0.66 µg·mL~(-1), respectively. The paclitaxel-loaded nano-micelles were potent in inhibiting cell migration as assessed by the scratch assay. PTX@FA-PLGA-NMs had good pro-apoptotic effect on cervical cancer HeLa cells and significantly promoted the uptake of HeLa cells. The results of in vitro experiments suggested that PTX@FA-PLGA-NMs could target and treat cervical cancer HeLa cells. Therefore, as nanodrug carriers, PTX@FA-PLGA-NMs with anti-cancer activity are a promising nano-system for improving the-rapeutic effects on tumors.


Assuntos
Antineoplásicos Fitogênicos , Neoplasias do Colo do Útero , Antineoplásicos Fitogênicos/farmacologia , Linhagem Celular Tumoral , Portadores de Fármacos , Feminino , Ácido Fólico , Glicolatos , Células HeLa , Humanos , Micelas , Paclitaxel , Tamanho da Partícula , Neoplasias do Colo do Útero/tratamento farmacológico
5.
Adv Physiol Educ ; 44(4): 726-733, 2020 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-33155832

RESUMO

Current interdisciplinary medical training calls for reforms and innovations in the assessment of pathophysiology education. Formative assessment is used to monitor student learning to provide ongoing feedback that can improve both learning and teaching. Beginning in 2016, we implemented a formative assessment composed of case-based multiple-choice questions (MCQs) for all students in all majors. In 2017, case study questions began to be employed in the formative assessment, and student-set, case-based questions were further introduced. Aiming to gather the students' suggestions and feedback on the mixed-method assessment, we conducted a survey on aspects such as the effectiveness of the assessment, assessment content and completion, opinions on student-set questions, and the impact on pathophysiology learning for students from 2017 to 2019. In addition, we compared students' semesterly final scores with those of previous students and evaluated the relationship between formative and summative assessment scores. The results for 1,277 students clearly showed that the reformed formative assessment system was well received by the students. The students thought that the formative assessment not only allowed for the provision of real-time feedback on the effectiveness of teaching and learning but also nurtured self-motivation, the development of analytical and problem-solving skills, and collaborative efforts. Both the semesterly final scores and the proportions of students scoring in higher score ranges increased after the implementation of the formative assessment, and the summative assessment scores were positively related to the formative assessment scores. Consequently, the reformed formative assessment system significantly improved the quality of pathophysiology education.


Assuntos
Educação de Graduação em Medicina , Estudantes de Medicina , Avaliação Educacional , Retroalimentação , Humanos , Inquéritos e Questionários
6.
Bone Joint Res ; 9(7): 368-385, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32793332

RESUMO

A balanced inflammatory response is important for successful fracture healing. The response of osteoporotic fracture healing is deranged and an altered inflammatory response can be one underlying cause. The objectives of this review were to compare the inflammatory responses between normal and osteoporotic fractures and to examine the potential effects on different healing outcomes. A systematic literature search was conducted with relevant keywords in PubMed, Embase, and Web of Science independently. Original preclinical studies and clinical studies involving the investigation of inflammatory response in fracture healing in ovariectomized (OVX) animals or osteoporotic/elderly patients with available full text and written in English were included. In total, 14 articles were selected. Various inflammatory factors were reported; of those tumour necrosis factor-α (TNF-α) and interleukin (IL)-6 are two commonly studied markers. Preclinical studies showed that OVX animals generally demonstrated higher systemic inflammatory response and poorer healing outcomes compared to normal controls (SHAM). However, it is inconclusive if the local inflammatory response is higher or lower in OVX animals. As for clinical studies, they mainly examine the temporal changes of the inflammatory stage or perform comparison between osteoporotic/fragility fracture patients and normal subjects without fracture. Our review of these studies emphasizes the lack of understanding that inflammation plays in the altered fracture healing response of osteoporotic/elderly patients. Taken together, it is clear that additional studies, preclinical and clinical, are required to dissect the regulatory role of inflammatory response in osteoporotic fracture healing. Cite this article: Bone Joint Res 2020;9(7):368-385.

7.
Genes (Basel) ; 11(5)2020 04 27.
Artigo em Inglês | MEDLINE | ID: mdl-32349370

RESUMO

Interleukin 8 (IL-8) participates in the immune response and has the function of inducing neutrophils to release lysosomal enzymes and eliminate pathogens. This study was to investigate the effect of single nucleotide mutations in the IL-8 gene promoter region on the coccidiosis resistance index. In this study, 180 infected Eimeria tenella (E. tenella) Jinghai yellow chickens were used as experimental samples. DNA sequencing technology was used to detect single nucleotide polymorphisms (SNPs) in the IL-8 gene promoter region. The association between these SNPs and coccidiosis resistance indexes (including superoxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GSH-PX), catalase (CAT), nitric oxide (NO), interleukin-1ß (IL-1ß), interleukin-2 (IL-2), interleukin-6 (IL-6), IL-8, and interferon-γ (IFN-γ)) were analyzed. Three SNPs (T-550C, G-398T, and T-360C) were detected. Significant associations were found between each genotype at the T-550C site with NO (p-value = 0.006) and IL-8 (p-value = 0.034) indexes. Significant associations were found between each genotype at the G-398T site with SOD (p-value = 0.042), CAT (p-value = 0.049), NO (p-value = 0.008), and IL-2 (p-value = 0.044) indexes. Significant associations were found between each genotype at the T-360C site with SOD (p-value = 0.007), NO (p-value = 0.046), IL-2 (p-value = 0.041), IL-8 (p-value = 0.039), and IFN-γ (p-value = 0.042) indexes. Haplotype analysis showed that multiple indexes of the H1H3 haplotype combination were significantly higher than other haplotype combinations. Therefore, mutation of the IL-8 gene promoter region has a significant regulatory effect on the coccidiosis resistance index, with a change in transcription factor binding potentially altering IL-8 gene expression, thereby further affecting the IL-8 level in plasma. However, the specific mechanism needs further study.


Assuntos
Galinhas/genética , Coccidiose/genética , Resistência à Doença/genética , Interleucina-8/genética , Animais , Galinhas/parasitologia , Coccidiose/parasitologia , Eimeria tenella/patogenicidade , Regulação da Expressão Gênica/genética , Polimorfismo de Nucleotídeo Único/genética , Regiões Promotoras Genéticas/genética
8.
Chem Commun (Camb) ; 55(74): 11123-11126, 2019 Sep 21.
Artigo em Inglês | MEDLINE | ID: mdl-31463500

RESUMO

A Pd-catalyzed decarboxylative cross-coupling of α,ß-unsaturated carboxylic acids with cyclic and acyclic epoxides has been developed. Both ß-monosubstituted and ß-disubstituted unsaturated carboxylic acids, as well as conjugated diene unsaturated carboxylic acids are suitable reaction substrates. Substituted homoallylic alcohols were obtained in moderate to good yields. The product was obtained as a mixture of diastereomers favoring the anti diastereomer of the cyclic epoxides. This work provides a method for the modification of complex organic molecules containing α,ß-unsaturated carboxylic acids.

9.
Zhongguo Zhong Yao Za Zhi ; 44(9): 1729-1733, 2019 May.
Artigo em Chinês | MEDLINE | ID: mdl-31342693

RESUMO

To establish a quality constant evaluation system of Alismatis Rhizoma decoction pieces,in order to provide reference for regulating the market circulation of this decoction pieces. A total of 18 batches of Alismatis Rhizoma decoction pieces were collected from different pharmaceutical factories,and the morphological parameters of each sample were tested. The content of alisol B 23-acetate in Alismatis Rhizoma decoction pieces was determined by HPLC in the 2015 edition of Chinese Pharmacopoeia,and the parameters such as quality constant and relative quality constant were calculated. The quality constant range of 18 batches of Alismatis Rhizoma decoction pieces was 0. 390-2. 076. If 18 batches of Alismatis Rhizoma decoction pieces were divided into 3 grades,taking 80% of the maximum quality constant as first grade,50% to 80% as second grade,and the rest as third grade,then the quality constant of firstgrade samples was ≥1. 66,the quality constant of second-grade samples was ≥1. 04 and <1. 66,and the quality constant of third-grade samples was <1. 04. The established quality constant evaluation method is objective and feasible,which can be used to classify the grade of Alismatis Rhizoma decoction pieces and provide a reference method to control the quality of this decoction pieces.


Assuntos
Alisma/química , Medicamentos de Ervas Chinesas/normas , Cromatografia Líquida de Alta Pressão , Controle de Qualidade , Rizoma/química
10.
Zhongguo Zhong Yao Za Zhi ; 44(6): 1273-1277, 2019 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-30989994

RESUMO

New Dihydroartemisinin Tablets were prepared,and a new dissolution determination method for the tablets was established,which provides reference for revising the quality standard.The dissolution experiment adopted the paddle method with 0.1 mol·L~(-1)hydrochloric acid solution 250 m L as the solvent,the rotating speed of 100 r·min~(-1)and the sampling time of 30 min.HPLC was adopted to determine the dissolution of dihydroartemisinin.The determination was performed on Agilent Eclipse XDB-C_(18)column with the mobile phase of acetonitrile-water(40∶60)at the flow rate of 1.0 m L·min~(-1).The detection wavelength was set at 216 nm,and the column temperature was 30℃.The sample size was 100µL.The linear range of dihydroartemisinin were 1.234 5-79.003 ng(r=1.000 0).The limit of quantitation was 0.308 6 ng,and the limit of detection was 0.154 3 ng.RSDs of precision tests were all lower than 1.0%.The recoveries were 98.09%-102.6%(RSD 1.8%,n=9).The average dissolutions of dihydroartemisinin in 3 batches of samples were 93.81%,92.61%,92.37%,respectively.The determination method is highly reproducible,accurate and reliable,and can objectively reflect the dissolution of Dihydroartemisinin Tablets,and provide a basis for revising the dissolution test of the current quality standard.Based on the dissolution rate,new tablets are superior to original tablets.


Assuntos
Artemisininas/química , Cromatografia Líquida de Alta Pressão , Solubilidade , Comprimidos
11.
Zhongguo Gu Shang ; 32(4): 383-386, 2019 Apr 25.
Artigo em Chinês | MEDLINE | ID: mdl-31027419

RESUMO

Total knee arthroplasty(TKA) has been the final clinical treatment of knee osteoarthritis at the final stage, postoperative limb and prosthesis alignment restoration directly affect clinical effect. In recent years, computer-assisted surgery has been used in TKA and obtained satisfied results. There paper has investigated that the use of computer-assisted systems could improve soft tissue balance after TKA, improve accuracy of installation of prosthesis, recover limb alignment and decrease intro-blood loss, postoperative fat embolism. Although computer-assisted navigation is not mainstream, with the continuous improvement of technology and innovation, the computer-assisted surgery could exert a more important role in TKA, and considerably beneficial effect on improvement of postoperative clinical effects.


Assuntos
Artroplastia do Joelho , Osteoartrite do Joelho , Cirurgia Assistida por Computador , Humanos , Articulação do Joelho , Período Pós-Operatório
12.
Zhongguo Gu Shang ; 32(2): 181-185, 2019 Feb 25.
Artigo em Chinês | MEDLINE | ID: mdl-30884939

RESUMO

Popliteal cyst is a common peri-knee cyst, also known as a Baker's cyst. With the current development of popliteal cysts, the pathogenesis is mainly due to increased pressure in the knee joint caused by various reasons, leading to a fluid-filled sac that can form behind the knee as a result of enlargement of the gastrocnemius-semimembranosus bursa. The current diagnostic methods include X-ray computed, ultrasound, and magnetic resonance. Among them, magnetic resonance imaging is considered the gold standard for the diagnosis of popliteal cysts. There are various treatments, including conservative treatment, traditional surgical resection and arthroscopic surgery. In recent years, the focus of the treatment of popliteal cysts has gradually shifted from the simple removal of cysts to arthroscopic treatment of intra-articular lesions and the treatment of joint-cyst communication, and achieved a good effect. However, there are still controversies about the most effective treatment, and the best surgery method is still uncertained. It is hoped that high quality prospective studies will be able to directly compare different surgery methods, so as to select the best treatment for popliteal cyst. This article reviews past literature research and describes in detail the epidemiology, pathological mechanism, clinical manifestations and signs, auxiliary examination, diagnosis and differential diagnosis and clinical treatment of popliteal cysts. Different diagnosis methods and treatment methods are compared and summarized to provide basis for clinical diagnosis and treatment.


Assuntos
Cisto Popliteal , Artroscopia , Bolsa Sinovial , Humanos , Articulação do Joelho , Cisto Popliteal/diagnóstico , Cisto Popliteal/terapia , Estudos Prospectivos
13.
Antioxid Redox Signal ; 31(1): 75-91, 2019 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-30829051

RESUMO

Aims: Vascular calcification is associated with cardiovascular death in patients with chronic kidney disease (CKD). Peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) plays an important role in various cardiovascular diseases. However, its role in vascular calcification remains unknown. Results: Adenine-induced rat CKD model was used to induce arterial medial calcification. The level of PGC-1α decreased in abdominal aorta of CKD rats. Overexpression of PGC-1α significantly ameliorated calcium deposition in rat abdominal aorta, isolated carotid rings, and cultured vascular smooth muscle cells (VSMCs). Mitochondrial reactive oxygen species (mtROS) increased in calcifying aorta and VSMCs. Upregulation of PGC-1α inhibited, whereas PGC-1α depletion promoted ß-glycerophosphate-induced mtROS production and calcium deposition. Moreover, PGC-1α increased superoxide dismutase 1 (SOD1) and SOD2 contents in vivo and in vitro, whereas SOD2 deletion eliminated PGC-1α-mediated mtROS change and promoted calcium deposition. Mechanistically, sirtuin 3 (SIRT3) expression declined in calcifying aorta and VSMCs, while PGC-1α overexpression restored SIRT3 expression. Inhibition of SIRT3 by 3-TYP or siRNA (small interfering RNA) reduced PGC-1α-induced upregulation of SOD1 and SOD2, and abolished the protective effect of PGC-1α on calcification of VSMCs. Importantly, PGC-1α was reduced in calcified femoral arteries in CKD patients. In phosphate-induced human umbilical arterial calcification, upregulation of PGC-1α attenuated calcium nodule formation, while this protective effect was abolished by SIRT3 inhibitor. Innovation: We showed for the first time that PGC-1α is an important endogenous regulator against vascular calcification. Induction of PGC-1α could be a potential strategy to treat vascular calcification in CKD patients. Conclusions: PGC-1α protected against vascular calcification by SIRT3-mediated mtROS reduction.


Assuntos
Mitocôndrias/metabolismo , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo/metabolismo , Insuficiência Renal Crônica/metabolismo , Sirtuína 3/metabolismo , Calcificação Vascular/metabolismo , Adenina/efeitos adversos , Idoso , Idoso de 80 Anos ou mais , Animais , Aorta Abdominal/metabolismo , Células Cultivadas , Modelos Animais de Doenças , Feminino , Regulação da Expressão Gênica , Humanos , Masculino , Músculo Liso Vascular/citologia , Músculo Liso Vascular/metabolismo , Estresse Oxidativo , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo/genética , Ratos , Insuficiência Renal Crônica/induzido quimicamente , Insuficiência Renal Crônica/complicações , Calcificação Vascular/genética
14.
Hepatobiliary Pancreat Dis Int ; 18(5): 458-463, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30522829

RESUMO

BACKGROUND: Hepatitis B virus covalently closed circular DNA (HBV cccDNA) is an important biomarker of hepatitis B virus infection. However, the current methods are not specific and sensitive. The present study aimed to develop a specific and sensitive assay method for the quantification of HBV cccDNA. METHODS: Exonuclease I (Exo I) & Exonuclease III (Exo III) and specific primer probes are used in real-time PCR. The virus particles isolated from peripheral blood mononuclear cells were used as negative control and HBV1.3 recombinant plasmid 3.2 kb circular DNA fragment was used as positive control. The methods of cccDNA detection were evaluated in cell lines, plasmid, animal model, patient serum and liver biopsies. RESULTS: A linear range of 101-107 copies/assay using specific primers for HBV cccDNA was established. HBV cccDNA were only detected in cell lines, animal model and liver tissue. It cannot be detected in serum samples. Intrahepatic HBV cccDNA level had good correlation with intrahepatic total HBV DNA level (r = 0.765, P < 0.001). CONCLUSIONS: The real-time quantitative PCR is an effective and feasible method for sensitive and specific detection of low copy number of cccDNA. The novel detection method is fast, provides high sensitivity and specificity and can be used in clinical practice.


Assuntos
DNA Circular/análise , Exodesoxirribonucleases , Vírus da Hepatite B/isolamento & purificação , Hepatite B/sangue , Reação em Cadeia da Polimerase em Tempo Real/métodos , Animais , Linhagem Celular , Modelos Animais de Doenças , Hepatite B/metabolismo , Vírus da Hepatite B/genética , Hepatócitos , Humanos , Fígado/metabolismo , Camundongos , Plasmídeos , Sensibilidade e Especificidade
15.
Chem Commun (Camb) ; 54(35): 4417-4420, 2018 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-29648563

RESUMO

The stereoselective synthesis of tri-substituted alkenes is challenging. Herein, we report a Ni-catalyzed regio- and stereo-selective hydroalkylation of internal alkynes with non-activated alkyl halides. This method does not use any sensitive organometallic reagents and shows good functional group compatibility, which enables the efficient synthesis of many tri-substituted olefins from readily available coupling partners. It also provides a straightforward method for the modification of bioactive organic molecules.

16.
J Mol Cell Cardiol ; 116: 135-144, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29438665

RESUMO

Cardiac fibrosis is characterized by excessive deposition of extracellular matrix (ECM) proteins in the myocardium and results in decreased ventricular compliance and diastolic dysfunction. Cartilage intermediate layer protein-1 (CILP-1), a novel identified cardiac matricellular protein, is upregulated in most conditions associated with cardiac remodeling, however, whether CILP-1 is involved in pressure overload-induced fibrotic response is unknown. Here, we investigated whether CILP-1 was critically involved in the fibrotic remodeling induced by pressure overload. Western blot analysis and immunofluorescence staining showed that CILP-1 was predominantly detected in cardiac myocytes and to a less extent in the interstitium. In isolated adult mouse ventricular myocytes and nonmyocytes, CILP-1 was found to be mainly synthesized by myocytes. CILP-1 expression in left ventricles was upregulated in C57BL/6 mice undergoing transverse aortic constriction (TAC). Myocardial CILP-1 knockdown aggravated whereas CILP-1 overexpression attenuated TAC-induced ventricular remodeling and dysfunction, as measured by echocardiography test, morphological examination, and gene expressions of fibrotic molecules. Incubation of cardiac fibroblasts with the conditioned medium containing full-length, N-terminal, or C-terminal CILP-1 inhibited transforming growth factor (TGF)-ß1-induced Smad3 phosphorylation and the subsequent profibrotic events. We first demonstrated that C-terminal CILP-1 increased Akt phosphorylation, promoted the interaction between Akt and Smad3, and suppressed Smad3 phosphorylation. Blockade of PI3K-Akt pathway attenuated the inhibitory effect of C-CILP-1 on TGF-ß1-induced Smad3 activation. We conclude that CILP-1 is a novel ECM protein possessing anti-fibrotic ability in pressure overload-induced fibrotic remodeling. This anti-fibrotic effect of CILP-1 attributes to interfering TGF-ß1 signaling through its N- and C- terminal fragments.


Assuntos
Miocárdio/metabolismo , Miocárdio/patologia , Pressão , Pirofosfatases/metabolismo , Transdução de Sinais , Fator de Crescimento Transformador beta1/metabolismo , Animais , Aorta/diagnóstico por imagem , Aorta/patologia , Aorta/fisiopatologia , Constrição Patológica , Dependovirus/metabolismo , Fibrose , Técnicas de Silenciamento de Genes , Camundongos Endogâmicos C57BL , Miócitos Cardíacos/metabolismo , Fosforilação , Ligação Proteica , Proteínas Proto-Oncogênicas c-akt/metabolismo , Pirofosfatases/genética , Proteína Smad3/metabolismo , Regulação para Cima/genética , Remodelação Ventricular
17.
Hepatol Res ; 48(3): E133-E145, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28707778

RESUMO

AIM: Chronic hepatitis B (CHB) patients with normal alanine aminotransferase (ALT) levels are not free from significant hepatic lesions. Recently, there has been an improved understanding of the clinical significance of quantitative hepatitis B core antibody levels (qAnti-HBc) during CHB management. In this cross-sectional study, we evaluated the utility of qAnti-HBc in identifying significant liver inflammation in CHB patients. METHODS: A total of 469 patients (training set, n = 363; validation set, n = 106) who underwent liver biopsy (LB) were included. The qAnti-HBc levels were quantified and the relationship between histology and serum markers was systematically analyzed. RESULTS: In the training set, qAnti-HBc levels were found to have significant diagnostic value for moderate to severe liver inflammation (≥G2) in all patients (area under the receiver operating characteristic curve [AUROC] = 0.768; 95% confidence interval [CI], 0.721-0.810; P < 0.001) and in patients with normal or near-normal ALT levels (AUROC = 0.767; 95% CI, 0.697-0.828; P < 0.001). Our novel index (AC index) for the identification of ≥G2 inflammation, which combined the qAnti-HBc and ALT levels, significantly improved diagnostic performance (AUROC = 0.813; 95% CI, 0.768-0.852) compared to the use of ALT alone (AUROC = 0.779; 95% CI, 0.732-0.821) in all patients. In the validation set, the AC index showed an improved AUROC of 0.890 (95% CI, 0.814-0.942) and 0.867 (95% CI, 0.749-0.943) in all patients and patients with normal ALT levels, respectively. CONCLUSIONS: The qAnti-HBc level predicts significant liver inflammation well, even in patients with normal or near-normal ALT levels. Compared with the conventional ALT level, the AC index is a more reliable non-invasive biomarker for significant liver inflammation in CHB patients.

18.
RSC Adv ; 8(72): 41561-41565, 2018 Dec 07.
Artigo em Inglês | MEDLINE | ID: mdl-35559282

RESUMO

Copper-catalyzed cross-coupling reactions of vinyl epoxide with arylboronates to obtain aryl-substituted homoallylic alcohols are described. The reaction selectivity was different to that of previously reported vinyl epoxide ring-opening reactions using aryl nucleophiles. The reaction proceeded under mild conditions, affording aryl-substituted homoallylic alcohols with high selectivity and in good to excellent yields. The reaction provides convenient access to aryl-substituted homoallylic alcohols from vinyl epoxide.

19.
Zhongguo Zhong Yao Za Zhi ; 42(10): 1819-1824, 2017 May.
Artigo em Chinês | MEDLINE | ID: mdl-29090536

RESUMO

As an outstanding representative of traditional Chinese medicine(TCM) prescriptions accumulated from famous TCM doctors' clinical experiences in past dynasties, classical TCM excellent prescriptions (cTCMeP) are the most valuable part of TCM system. To support the research and development of cTCMeP, a series of regulations and measures were issued to encourage its simplified registration. There is still a long-way to go because many key problems and puzzles about technology, registration and administration in cTCMeP R&D process are not resolved. Based on the analysis of registration and management regulations of botanical drug products in FDA of USA and Japan, and EMA of Europe, the possible key problems and countermeasures in chemistry, manufacture and control (CMC) of simplified registration of cTCMeP were analyzed on the consideration of its actual situation. The method of "reference decoction extract by traditional prescription" (RDETP) was firstly proposed as standard to evaluate the quality and preparation uniformity between the new developing product under simplified registration and traditional original usages of cTCMeP, instead of Standard Decoction method in Japan. "Totality of the evidence" approach, mass balance and bioassay/biological assay of cTCMeP were emphatically suggested to introduce to the quality uniformity evaluation system in the raw drug material, drug substance and final product between the modern product and traditional decoction.


Assuntos
Medicamentos de Ervas Chinesas/normas , Preparações Farmacêuticas/normas , Controle de Qualidade , Medicina Tradicional Chinesa
20.
Zhongguo Zhong Yao Za Zhi ; 42(7): 1331-1337, 2017 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-29052395

RESUMO

To optimize the purification process of gynostemma pentaphyllum saponins (GPS) based on "adjoint marker" online control technology with GPS as the testing index. UPLC-QTOF-MS technology was used for qualitative analysis. "Adjoint marker" online control results showed that the end point of load sample was that the UV absorbance of effluent liquid was equal to half of that of load sample solution, and the absorbance was basically stable when the end point was stable. In UPLC-QTOF-MS qualitative analysis, 16 saponins were identified from GPS, including 13 known gynostemma saponins and 3 new saponins. This optimized method was proved to be simple, scientific, reasonable, easy for online determination, real-time record, and can be better applied to the mass production and automation of production. The results of qualitative analysis indicated that the "adjoint marker" online control technology can well retain main efficacy components of medicinal materials, and provide analysis tools for the process control and quality traceability.


Assuntos
Medicamentos de Ervas Chinesas/química , Gynostemma/química , Saponinas/isolamento & purificação , Biomarcadores , Cromatografia Líquida de Alta Pressão , Espectrometria de Massas
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