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Objective:To investigate the distribution of intraocular pressure (IOP) in high-altitude population aged 18 years and over in Xining, Qinghai and establish the reference interval (RI) of IOP.Methods:A cross-sectional study was conducted in Xining, Qinghai Province at 2.271 km above sea level from September 2019 to May 2020.Ophthalmic examinations and IOP measurement were conducted among subjects from Physical Examination Center of Qinghai Provincial People's Hospital.The subjects who had been living in Xining without leaving for three months were enrolled.Ophthalmic examinations included vision examination, IOP measurement, slit-lamp microscopy, fundus photography, anterior and posterior segment optical coherence tomography.IOP was measured using Goldmann applanation tonometry under local anesthesia.Subjects with factors that could cause significant changes in IOP and affect the accuracy of IOP measurement, and those who were unable to receive IOP measurement were excluded.Subjects were grouped according to sex, age and ethnicity, and the distribution and RI of IOP were compared among all groups.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Beijing Tongren Hospital, Capital Medical University (No.TRECKY2017-024). Written informed consent was obtained from each subject.Results:A total of 6 120 subjects (6 120 eyes) aged 18-90 years old were enrolled, including 2 850 males and 3 270 females with average age of (45.54±13.85) years.The average IOP of high-altitude population in Xining, Qinghai Province was (14.32±1.93) mmHg (1 mmHg=0.133 kPa), with the RI of 10.54-18.10 mmHg.The average IOP was (14.42±1.98) mmHg in male with the RI of 10.54-18.30 mmHg, (14.23±1.88) mmHg in female with the RI of 10.55-17.91 mmHg.The IOP of male was higher than that of female ( t=3.71, P<0.001). The IOP of Han, Tibetan, Hui and other nationalities were (14.38±1.91), (13.93±2.06), (14.21±1.87), (13.94±1.95) mmHg, respectively, with a statistically significant overall difference ( F=6.73, P<0.001). The IOP of Han nationality was significantly higher than that of Tibetan, Hui and other nationalities, and the differences were statistically significant (all at P<0.05). Conclusions:RI of IOP in high-altitude population from Xining, Qinghai is lower compared with normal altitude area.
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The participants in this study were 20-49 years old rural childbearing age people who received the National Free Preconception Health Examination Project (NFPHEP) in Yunnan Province during 2013 to 2019. The proportion of ABO and RhD blood groups among different ethnic groups and different areas were calculated. The proportion of 2 748 131 participants with blood group A phenotype was highest (32.60%), followed by O (30.60%), B (27.33%) and AB (9.47%). In the RhD blood system, the proportion of the RhD positivity (RhD+) and RhD negativity (RhD-) group were 99.29% and 0.71% respectively. The proportions blood groups were significantly different among ethnic groups and areas (all P<0.001). Among 18 ethnic groups with more than 3 000 participants, Yao (42.75%), Bouyei (40.58%) and Dai (40.37%) ethnic groups had higher proportion of blood group O phenotype than other ethnic groups. Wa ethnic groups had highest proportion of the A (40.15%) and AB phenotypes (11.23%). Miao ethnic group (34.70%) and Lahu ethnic group (34.42%) had higher proportion of blood group B phenotype than other ethnic groups. Wa ethnic group had the highest proportion of RhD-group (1.88%). In all 16 prefectures of Yunnan, the proportion of blood group O phenotype was highest in Xishuangbanna Dai Autonomous Prefecture (40.27%). Baoshan city (36.39%), Lincang city (36.22%) and Dali Bai autonomous prefecture (36.06%) had higher proportion of blood group A phenotype than other regions. Diqing Tibetan Autonomous Prefecture (30.83%) and Qujing city (30.48%) had higher proportion of blood group B phenotype than other areas, while Zhaotong city had a highest proportion of blood group AB phenotype (11.19%). The proportion of RhD-group was highest in Honghe hani and Yi nationality autonomous prefecture(1.37%). The A RhD+(39.36%), A RhD-(0.78%), AB RhD+(11.03%), AB RhD-(0.20%) and O RhD-(0.48%) blood groups were higher proportion in Wa ethnic group than in other ethnic groups (P<0.001).
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Adult , Humans , Middle Aged , Young Adult , Blood Group Antigens , China , Ethnicity , Rural PopulationABSTRACT
Objective:To prepare camel-derived nanoantibodies that can bind to the recombinant protein VirB12 antigen with high affinity and lay the foundation for further research.Methods:Xinjiang Bactrian camels were immunized six times with VirB12 recombinant protein, total RNA was extracted from lymphocytes isolated from peripheral blood, and the VHH gene fragment was amplified by nested PCR to construct a phage VHH display library. ELISA solid-phase affinity and enrichment methods were used for screening. After three rounds of affinity screening, the clones enriched in the second and third rounds were randomly picked out, and the binding of a nanoantibody with soluble expression to VirB12 was analyzed by ELISA. After sequence determination and multiple alignment, repetitive sequences were removed, and finally five non-redundant sequences were obtained, which were named D1, E6, H8, H9, and H10. The five identified nanoantibody genes were transformed into the WK6 strain, and the soluble expression of an intercellular substance was carried out at 16 °C. After purified expression of Ni-NTA, the binding ability and thermal stability of nanoantibodies and the antigen VirB12 protein were detected by Western Blot and ELISA.Results:Five strains of nanoantibodies were expressed in WK6 bacteria in soluble form. SDS-PAGE showed that the purity of five anti-VirB12 nanoantibodies was close to 90%, and they had high antigen-binding activity and obvious antigen-antibody concentration dependence. All four strains of nanoantibodies showed high thermal stability, and after being treated at 90 ℃, they could still retain more than 60% binding activity.Conclusions:In the study, a VHH phage display library with a capacity of 2.8×10 8 cfu/ml was constructed from Xinjiang Bactrian camel lymphocytes immunized with VirB12 recombinant protein. Five anti-VirB12 nanoantibodies with high affinity and thermal stability were obtained through solid-phase screening and enrichment and soluble monoclonal ELISA detection. These results laid the foundation for further development of VirB12 nanoantibodies.
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Objective:To established a method for the detection of soluble programmed death ligand 1 (PD-L1) protein in serum based on the poly nanoantibody of lumazine synthase(LS).Methods:A dual nanobody-based sandwich ELISA was established with a competitive ELISA to screen nanobodies recognizing different epitopes of PD-L1 as paired antibodies. To improve sensitivity, PD-L1 nanobody P3C8 and lumazine synthase(LS) were fused, and nanobodies were obtained in polymeric forms as sPD-L1 protein captures, so as to develop an LS-displayed polymeric nanobody-based sandwich ELISA (LSNbs-ELISA) method to detect sPD-L1.Results:Compared with the Nbs-ELISA method, the LSNbs-ELISA method is approximately 11-fold more sensitive for sPD-L1 detection. The limit of detections (LODs) of Nbs-ELISA and LSNbs-ELISA for sPD-L1 in serum were 2.87 ng/ml and 0.255 ng/ml, respectively. Both assays were highly specific for the detection of sPD-L1 and did not react with structure-related proteins PD-1, CD27, CD70, CD137, and CD147 when spiked into the human serum.Conclusions:The Nbs-ELISA and LSNbs-ELISA assays both have high sensitivity and specificity for detecting sPD-L1 in serum and could have potential clinical applications.
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Objective:To evaluate the potential of a previously identified CDR3 only single-domain antibodies (sdAbs) fragment, NBL42, as a general framework for affinity transfer.Methods:The H3 loops of VHH-A4(A4), VHH-H5(H5), cAb-Lys3(L3) and B6H12 which bind with alliinase, PD-1, lysozyme and CD47, respectively, were grafted into the corresponding loop of NBL42. The genes of the reconstituted CDR3 only sdAbs were synthesized, expressed in E. coliand purified with Ni 2+ column affinity chromatography. The antigen binding and stability of the recombinant CDR3 only sdAbs were assayed by ELISA. Results:The recombinant NBL42-A4CDR3, NBL42-H5CDR3, NBL42-L3CDR3 and NBL42-B6H12CDR3 ran as a single peak at 15, 15, 28 and 16 kDa, respectively, in SDS-PAGE as expected molecular weight. Grafted sdAbs NBL42-A4CDR3 and NBL42-H5CDR3 expressed in a soluble form and specifically bind with alliinase and PD-1, respectively, but lost about 50% of their binding activity. In contrast, the grafted sdAbs NBL42-Lys3CDR3 and NBL42-B6H12CDR3 completely lost their antigen binding capacity. NBL42 sdAbs and grafted sdAbs NBL42-A4CDR3 and NBL42-H5CDR3 retain roughly half of their binding activity after 90 ℃ heat treatment, indicating high stability. The C88Y mutation in NBL42 and the Swiss Mode 3D model predicted that the C88Y residue in FR3 may play a key role in NBL42 stability and CDR3 affinity transfer.Conclusions:The structure of NBL42 has potential as a framework for CDR3 transplantation and affinity transfer.
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In this study, a method for simultaneous quantitative analysis of 6 salvianolic acids and 4 tanshinones in extracts of Salviae Miltiorrhizae Radix et Rhizoma was established by ultra-high performance liquid chromatography (UHPLC). The semi-biomimetic method was applied to simulate digestion process in vitro, to explore the digestion and transport characters of oral administration through the gastrointestinal tract, and to explain the content ratio changes and bioaccessibility of active ingredients in Salviae Miltiorrhizae Radix et Rhizoma. The results showed that the 10 index components have a good linear relationship in the corresponding concentration range, and the average recovery rate was 91.35% to 105.65%. After simulated digestion in vitro, types of chemical composition in simulated gastric fluid and simulated intestinal fluid digested extracts of Salviae Miltiorrhizae Radix et Rhizoma did not change significantly. While the content ratio of salvianolic acid B and rosmarinic acid decreased, and the content ratio of protocatechuic aldehyde and danshensu increased. In the simulated gastric fluid digestion extract of Salviae Miltiorrhizae Radix et Rhizoma, the order of bioaccessibility was: danshensu (50.19%) > salvianolic acid B (33.44%) > lithospermic acid (27.34%) > salvianolic acid A (21.71%) > rosmarinic acid (12.31%). In the simulated intestinal fluid digestion extract of Salviae Miltiorrhizae Radix et Rhizoma, the order of bioaccessibility was: 15,16-dihydrotanshinone Ⅰ (5.45%) > tanshinone Ⅰ (3.67%) > cryptotanshinone (3.29%) > tanshinone ⅡA (3.01%) > salvianolic acid A (2.39%) > lithospermic acid (1.57%) > salvianolic acid B (1.02%) > danshensu (0.41%) > rosmarinic acid (0.34%). In conclusion, the UHPLC method established in this study can be applied for accurately and sensitively detecting the contents of 6 salvianolic acids and 4 tanshinones in Salviae Miltiorrhizae Radix et Rhizoma. The results of semi-biomimetic extraction showed that not all components were extracted with simulated gastric fluid and simulated intestinal fluid, especially rosmarinic acid and salvianolic acid B. Therefore, in the quality study of Salviae Miltiorrhizae Radix et Rhizoma and its extract, bioavailability should be considered at the same time when select quality markers and determine their content limits.
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Objective:To investigate the clinical values of the differences between hematocrit and serum albumin (HCT-ALB) for evaluating the severity of patients with acute paraquat (PQ) poisoning.Methods:Patients with acute PQ poisoning admitted to the Second People's Hospital of Yunnan Province from January 2018 to December 2019 were enrolled, and healthy voluteers during the same period were selected as the control. The general information, poisoning dose and poisoning time of patients, as well as the HCT and serum ALB levels before blood product infusion, intravenous infusion, or hemopurification at admission were collected, and the HCT-ALB was calculated. According to the results of rapid semiquantitative test of PQ in urine at admission, the patients were divided into PQ low concentration group (0-10 mg/L) and PQ high concentration group (30-100 mg/L). The relationship between poisoning time, poisoning dose, HCT-ALB and the degree of acute PQ poisoning were analyzed, and Spearman method was used to analyze the grade correlation.Results:A total of 295 patients with acute PQ poisoning were enrolled, including 118 cases in PQ low concentration group and 177 cases in PQ high concentration group, and another 200 healthy persons matched with PQ patients in gender and age (healthy control group). The poisoning time of PQ low concentration group was significantly longer than that of PQ high concentration group [hours: 11.0 (6.0, 60.0) vs. 8.0 (5.0, 20.5), P < 0.01], but the poisoning dose was significantly lower than that of PQ high concentration group [mL: 10.0 (5.8, 15.0) vs. 40.0 (20.0, 80.0), P < 0.01]. The HCT and HCT-ALB in PQ low and high concentration groups were significantly higher than those of the healthy control group [HCT: (43.14±4.41)%, (43.54±5.40)% vs. (42.14±2.15)%, HCT-ALB: 3.59±6.26, 5.94±7.80 vs. -7.26±3.55, all P < 0.01], but ALB was significantly lower than that of the healthy control group (g/L: 39.54±5.74, 37.60±7.15 vs. 49.40±3.41, both P < 0.01). With the increase of urine PQ concentration, the HCT and HCT-ALB further increased, and ALB further decreased. There were significant differences between PQ high concentration group and PQ low concentration group [HCT: (43.54±5.40)% vs. (43.14±4.41)%, HCT-ALB: 5.94±7.80 vs. 3.59±6.26, ALB (g/L): 37.60±7.15 vs. 39.54±5.74, all P < 0.05]. The poisoning severity of patients with acute PQ poisoning were negatively correlated with poisoning time and ALB ( r values were -0.195 and -0.695, respectively, both P < 0.01), there were positively correlated with poisoning dose, HCT, and HCT-ALB ( r values were 0.650, 0.256, 0.737, respectively, all P < 0.01), and the correlation between HCT-ALB and poisoning severity was the strongest. Conclusion:The HCT-ALB can reflect the poisoning severity of patients with acute PQ poisoning and indirectly reveal the pathological changes of microvessels in patients with acute PQ poisoning.
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Biomarkers are defined as a characteristic that is objectively measured and evaluated as an indicator of normal biological processes, pathogenic processes, or pharmacological responses to a therapeutic intervention. Biomarkers can help the decision-making process for new drug research and development, provide guidance for the early clinical development of candidate drugs and reduce the risk of failure. Therefore, as a key factor in the development of new drugs, the discovery and research on biomarkers has increased the interest of the pharmaceutical industry and regulatory agencies. Guidelines on the development and use of biomarkers have been issued by drug regulatory agencies including the EMA, FDA and ICH. Biomarkers are encouraged to be used to facilitate drug development by these relevant regulatory agencies, and also to be used to monitor the safety and efficacy of drugs in post-marketing drug surveillance. The application of biomarkers is encouraged at different stages of a drug's life cycle, including at the stage of basic science research and target identification, prototype design or discovery, preclinical development, clinical development, FDA filling/approval and launch, as well as post-marketing was reviewed. The identification, development, and application of biomarkers in pharmaceutical research is discussed.
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Proton nuclear magnetic resonance(~1H-NMR) is used to investigate the effect of Renshenjian Decoction on serum and urine metabolism of type 2 diabetic rats with insulin resistance induced by high-sugar and high-fat diet combined with low-dose streptozotocin(STZ). After the successful establishment of the insulin resistance model of type 2 diabetes, administration for 35 days, the serum and urine of rats were taken. Once the ~1H-NMR data have been collected and processed, PCA and OPLS-DA were used to analyze them. The results show that: compared with the blank group, the contents of methionine, taurine, α-glucose and β-glucose in the serum of the model group increased significantly(P<0.001), while the contents of 3-hydroxybutyric acid, lactic acid and unsaturated fatty acids decreased significantly(P<0.01). In the model group, the contents of trimethylamine oxide, glycine, α-glucose, β-glucose, taurine and phosphocholine in urine increased significantly(P<0.05), while the contents of creatine, lactic acid, acetic acid and citric acid decreased significantly(P<0.05). Compared with the model group, the contents of 3-hydroxybutyric acid and unsaturated fatty acids in serum of rats in the treatment group increased significantly(P<0.05), while the contents of taurine, α-glucose and β-glucose decreased significantly(P<0.01). In the treatment group, the contents of lactic acid, taurine and creatine in urine increased significantly(P<0.05), while the contents of trimethylamine oxide, glycine, α-glucose, β-glucose and phosphocholine decreased significantly(P<0.01). The results show that Renshenjian Decoction can regulate metabolic disorder and promote the metabolic phenotype to return to the normal range. It displayed therapeutic effect on type 2 diabetic rats with insulin resistance and provided a certain scientific basis for the biological basic research of Renshenjian Decoction by improving insulin resistance in diabetes mellitus.
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Animals , Rats , Blood Glucose , Diabetes Mellitus, Experimental , Diabetes Mellitus, Type 2 , Metabolomics , Proton Magnetic Resonance Spectroscopy , Rats, Sprague-DawleyABSTRACT
@#Organelles have their special functions, they interact with each other and coordinate a series of important physiological functions at the same time. Organelle interaction occurs at membrane contact sites(MCSs), where the membranous organelle endoplasmic reticulum is the core, and specific tethered proteins at the membrane contact site bind to the organelle membrane and various protein complexes work together to perform specific functions, such as lipid transport, Ca2+ transfer, etc. This review studies on the structure and function of membrane contact sites and their key roles in organelle interactions, focusing on the connection between the endoplasmic reticulum and plasma membrane, mitochondria and Golgi, as well as the association between the key proteins at membrane contact sites and the occurrence and development of various diseases.
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Background@#Collagen type IV (COL4)-related nephropathy includes a variety of kidney diseases that occur with or without extra-renal manifestations caused by COL4A3-5 mutations. Previous studies revealed several novel mutations, including three COL4A3 missense mutations (G619R, G801R, and C1616Y) and the COL4A3 chr:228172489delA c.4317delA p.Thr1440ProfsX87 frameshift mutation that resulted in a truncated NC1 domain (hereafter named COL4A3 c.4317delA); however, the mutation mechanisms that lead to podocyte injury remain unclear. This study aimed to further explore the mutation mechanisms that lead to podocyte injury.@*Methods@#Wild-type (WT) and four mutant COL4A3 segments were constructed into a lentiviral plasmid, then stably transfected into human podocytes. Real-time polymerase chain reaction and Western blotting were applied to detect endoplasmic reticulum stress (ERS)- and apoptosis-related mRNA and protein levels. Then, human podocytes were treated with MG132 (a proteasome inhibitor) and brefeldin A (a transport protein inhibitor). The human podocyte findings were verified by the establishment of a mus-Col4a3 knockout mouse monoclonal podocyte using clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) technology.@*Results@#Our data showed that COL4A3 mRNA was significantly overexpressed in the lentivirus stably transfected podocytes. Moreover, the COL4A3 protein level was significantly increased in all groups except the COL4A3 c.4317delA group. Compared to the other test groups, the COL4A3 c.4317delA group showed excessive ERS and apoptosis. Podocytes treated with MG132 showed remarkably increased intra-cellular expression of the COL4A3 c.4317delA mutation. MG132 intervention improved higher ERS and apoptosis levels in the COL4A3 c.4317delA group. Mouse monoclonal podocytes with COL4A3 chr:82717932insA c.4852insA p.Arg1618ThrfsX4 were successfully acquired; this NC1-truncated mutation suggested a higher level of ERS and relatively remarkable level of apoptosis compared to that of the WT group.@*Conclusions@#We demonstrated that excessive ERS and ERS-induced apoptosis were involved in the podocyte injury caused by the NC1-truncated COL4A3 mutation. Furthermore, proteasome pathway intervention might become a potential treatment for collagen type IV-related nephropathy caused by a severely truncated COL4A3 mutation.
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BACKGROUND@#Collagen type IV (COL4)-related nephropathy includes a variety of kidney diseases that occur with or without extra-renal manifestations caused by COL4A3-5 mutations. Previous studies revealed several novel mutations, including three COL4A3 missense mutations (G619R, G801R, and C1616Y) and the COL4A3 chr:228172489delA c.4317delA p.Thr1440ProfsX87 frameshift mutation that resulted in a truncated NC1 domain (hereafter named COL4A3 c.4317delA); however, the mutation mechanisms that lead to podocyte injury remain unclear. This study aimed to further explore the mutation mechanisms that lead to podocyte injury.@*METHODS@#Wild-type (WT) and four mutant COL4A3 segments were constructed into a lentiviral plasmid, then stably transfected into human podocytes. Real-time polymerase chain reaction and Western blotting were applied to detect endoplasmic reticulum stress (ERS)- and apoptosis-related mRNA and protein levels. Then, human podocytes were treated with MG132 (a proteasome inhibitor) and brefeldin A (a transport protein inhibitor). The human podocyte findings were verified by the establishment of a mus-Col4a3 knockout mouse monoclonal podocyte using clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) technology.@*RESULTS@#Our data showed that COL4A3 mRNA was significantly overexpressed in the lentivirus stably transfected podocytes. Moreover, the COL4A3 protein level was significantly increased in all groups except the COL4A3 c.4317delA group. Compared to the other test groups, the COL4A3 c.4317delA group showed excessive ERS and apoptosis. Podocytes treated with MG132 showed remarkably increased intra-cellular expression of the COL4A3 c.4317delA mutation. MG132 intervention improved higher ERS and apoptosis levels in the COL4A3 c.4317delA group. Mouse monoclonal podocytes with COL4A3 chr:82717932insA c.4852insA p.Arg1618ThrfsX4 were successfully acquired; this NC1-truncated mutation suggested a higher level of ERS and relatively remarkable level of apoptosis compared to that of the WT group.@*CONCLUSIONS@#We demonstrated that excessive ERS and ERS-induced apoptosis were involved in the podocyte injury caused by the NC1-truncated COL4A3 mutation. Furthermore, proteasome pathway intervention might become a potential treatment for collagen type IV-related nephropathy caused by a severely truncated COL4A3 mutation.
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Ethnomedicine is the precious wealth left by ethnic minorities in their struggle against diseases. It is similar to traditional Chinese medicine in a narrow sense and has the characteristics of multi-component,multi-target and multi-channel synergy. Under the guidance of the theory of ethnomedicine,the combination of ethnomedicine and network pharmacology will help to understand the essence of the prevention and treatment of ethnomedicines in a dynamic and holistic manner. This paper reviews the research progress of network pharmacology applied in ethnomedicine,analyses the problems and challenges existing in the application of network pharmacology in ethnomedicine research at present,such as inaccurate data and information,lack of network analysis platform for effective analysis of dose-effect relationship of chemical constituents and weak basic research of ethnomedicine,and puts forward corresponding prospects.
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Ethnopharmacology , Medicine, Chinese Traditional , Medicine, TraditionalABSTRACT
Primary glomerulonephritis (PGN) remains the major cause of end-stage renal disease (ESRD) in our country. The histologic entity of PGN mainly includes immunoglobulin A nephropathy (IgAN), idiopathic membranous nephropathy (IMN), minimal change disease (MCD), focal and segmental glomerulosclerosis (FSGS) and membranoproliferative glomerulonephritis (MPGN). The pathogenesis of PGN is correlated with renal immune complex deposition, podocyte injury, infection and abnormal regulation of complement system. Nowadays PGN is short of specific treatments, the main therapeutic methods of PGN consists of renin angiotensin aldosterone system (RAAS) inhibitor, corticosteroids, cytotoxic drugs, lipid-lowering agents, anticoagulant therapy and antiplatelet adhesion. Patients who are drug-resistant or intolerance of the side effects will have a poor prognosis. Rituximab (RTX) is a chimeric monoclonal anti-CD20 antibody. The binding of RTX to CD20 on the cell membrane of B lymphocytes leads to significant depletion of peripheral B lymphocytes, which plays an immunosuppressive role. Rituximab is originally approved for the treatment of lymphoma, after that there was growing evidence showed RTX was effective in part of immunological diseases, including systemic lupus erythematosus and anitneutrophil cytoplasmic antibody associated vasculitis. As a result, whether RTX will act as an effective treatment modality in PGN has aroused extensive attention. In recently years, clinical researches concerning RTX used for the treatment of PGN have been published in succession. This paper reviewed clinical studies focused on the use of rituximab in the treatment of IMN, MCD, FSGS and IgAN.
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Melatonin has been reported to inhibit hepatic fibrosis and the mechanism may be correlated to its anti-oxidant effect.Nevertheless,the mechanism is not completely identified.This study was conducted to investigate the effects of melatonin on TGF-β1/Smad signaling pathway in liver fibrosis in rats.The liver fibrosis model was made by the subcutaneous injection of CCl4.The liver pathology changes were detected using hematoxylin and eosin (H&E) staining and Van Gieson (VG) staining.Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities were measured with an autoanalyzer.Glutathione peroxidase (GPx) activities and levels of malondialdehyde (MDA) and hydroxyproline (Hyp) in liver were evaluated by spectrophotometry.Expression levels of TGF-β1,Smad2/3,phosphorylated Smad2/3 (p-Smad2/3) and Smad7 in liver were detected by immunohistochemistry and Western blot analysis.Results showed that melatonin suppressed CC14-induced liver fibrosis,along with an improvement in histological changes,significant decreases in pathologic grading sores and obvious decreases in Hyp levels in liver.Melatonin improved liver function indicated by decreased serum ALT and AST activities.In addition,melatonin exerted its anti-oxidant effects,as supported by decreased MDA levels and increased GPx activities in liver.Furthermore,melatonin inhibited TGF-β1/Smad pathway,as evidenced by decreased TGF-β1,Smad2/3 and p-Smad2/3 expression and increased Smad7 expression in liver.In conclusion,melatonin may suppress CCl4-induced hepatic fibrosis in rats via inhibiting TGF-β1/Smad pathway.It is possible for melatonin to be a potential reagent to treat and cure liver fibrosis.
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Melatonin has been reported to inhibit hepatic fibrosis and the mechanism may be correlated to its anti-oxidant effect.Nevertheless,the mechanism is not completely identified.This study was conducted to investigate the effects of melatonin on TGF-β1/Smad signaling pathway in liver fibrosis in rats.The liver fibrosis model was made by the subcutaneous injection of CCl4.The liver pathology changes were detected using hematoxylin and eosin (H&E) staining and Van Gieson (VG) staining.Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities were measured with an autoanalyzer.Glutathione peroxidase (GPx) activities and levels of malondialdehyde (MDA) and hydroxyproline (Hyp) in liver were evaluated by spectrophotometry.Expression levels of TGF-β1,Smad2/3,phosphorylated Smad2/3 (p-Smad2/3) and Smad7 in liver were detected by immunohistochemistry and Western blot analysis.Results showed that melatonin suppressed CC14-induced liver fibrosis,along with an improvement in histological changes,significant decreases in pathologic grading sores and obvious decreases in Hyp levels in liver.Melatonin improved liver function indicated by decreased serum ALT and AST activities.In addition,melatonin exerted its anti-oxidant effects,as supported by decreased MDA levels and increased GPx activities in liver.Furthermore,melatonin inhibited TGF-β1/Smad pathway,as evidenced by decreased TGF-β1,Smad2/3 and p-Smad2/3 expression and increased Smad7 expression in liver.In conclusion,melatonin may suppress CCl4-induced hepatic fibrosis in rats via inhibiting TGF-β1/Smad pathway.It is possible for melatonin to be a potential reagent to treat and cure liver fibrosis.
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OBJECTIVE: To determine the feasibility of reduced field-of-view diffusion-weighted imaging (rFOV DWI) with multi-b values to detect functional variability in transplanted kidneys. MATERIALS AND METHODS: Using a 3T MRI scanner, multi-b rFOV DWI of transplanted kidney or native kidney was performed in 40 renal transplantation recipients and 18 healthy volunteers. The patients were stratified, according to an estimated glomerular filtration rate (eGFR): Group 1, eGFR ≥ 60 mL/min/1.73 m2; Group 2, eGFR ≥ 30 mL/min/1.73 m2 and < 60 mL/min/1.73 m2; Group 3, eGFR < 30 mL/min/1.73 m2. Total apparent diffusion coefficient (ADCT), perfusion-free ADC (ADCD) and perfusion fraction (FP) of kidneys were calculated and compared among the four groups. Correlations between the imaging results and eGFR were assessed. RESULTS: All volunteers had eGFR ≥ 60 mL/min/1.73 m2, while 16, 16, and 8 patients were included in Groups 1, 2, and 3, respectively. In the renal cortex, ADCT was higher in Group 1 ([1.65 ± 0.13] × 10−3 mm2/s) than Group 3 ([1.44 ± 0.11] × 10−3 mm2/s) (p < 0.05), and the inter-group differences of FP values were significant (all p < 0.05) (0.330 ± 0.024, 0.309 ± 0.019, 0.278 ± 0.033, and 0.250 ± 0.028 for control group, Groups 1, 2, and 3, respectively). Renal cortical ADCT, ADCD, FP, and renal medullary ADCT and FP correlated positively with eGFR (r = 0.596, 0.403, 0.711, 0.341, and 0.323, respectively; all p < 0.05). When using 0.278 as the cutoff value, renal cortical FP had a sensitivity of 97.1% and a specificity of 66.7% for predicting decreased renal function. CONCLUSION: Multi-b rFOV DWI presents transplanted kidneys with high resolution, which is a promising functional tool for non-invasively monitoring function of transplanted kidneys.
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Humans , Diffusion , Glomerular Filtration Rate , Healthy Volunteers , Kidney Transplantation , Kidney , Magnetic Resonance Imaging , Perfusion , Sensitivity and Specificity , Transplantation , VolunteersABSTRACT
Objective To investigate the protective effect of thalidomide on acute lung injury (ALI) induced by paraquat (PQ) poisoning in rats and its possible mechanism. Methods Sixty SPF Wistar rats were randomly divided into six groups with 10 rats in each group. The rat model of PQ poisoning was reproduced by intraperitoneal injection of PQ solution 20 mg/kg (PQ model group), and the rats were treated by intraperitoneal injection of gradient thalidomide (50, 100, 200 mg/kg treatment groups) 30 minutes later continuously for 3 days. The normal saline (NS) control group and thalidomide control group (thalidomide 200 mg/kg) were established. After 3 days, the abdominal aorta blood was collected, and the superoxide dismutase (SOD) activity was determined by hydroxylamine method, serum malondialdehyde (MDA) content was determined by thiobarbituric acid method. The rats were sacrificed for lung tissue, the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were determined by enzyme-linked immunosorbent assay (ELISA). The phosphorylation levels of p65 and inhibitor-α of nuclear factor-κB (NF-κB) (IκB-α), which were the NF-κB signaling pathway proteins, were determined by Western Blot. The pathological changes in lung tissue were observed under light microscope by hematoxylin-eosin (HE) staining. Results Under microscope, obvious congestion of pulmonary interstitial and alveolar septum, a large number of inflammatory cells infiltration and thickened alveolar wall were observed after 3 days of PQ poisoning, and the congestion of pulmonary interstitial and alveolar septum, edema and inflammatory cells infiltration in the lung tissue were significantly reduced after treatment of 50, 100, 200 mg/kg thalidomide, but compared with NS control group, there was still a small amount of edema fluid, inflammatory cells and erythrocytes in the lungs tissue. Compared with the NS control group, serum MDA content and the levels of TNF-α and IL-6, and the phosphorylation of p65 and IκB-α in lung tissue were significantly increased after PQ exposure, and the activity of serum SOD was significantly decreased. Treatment with 50, 100, 200 mg/kg thalidomide could significantly reduce the levels of MDA, TNF-α, IL-6, and phosphorylation of IκB-α and p65, and increase SOD activity, in a dose-dependent manner, and the levels were significantly different from PQ model group [MDA (mmol/L): 8.26±1.20, 6.72±1.18, 5.51±1.44 vs. 9.02±1.03, TNF-α (ng/mg): 3.00±0.14, 1.84±0.18, 1.58±0.11 vs. 3.30±0.14, IL-6 (ng/mg): 1.26±0.04, 1.06±0.04, 0.97±0.08 vs. 1.97±0.07, p-p65/p65: 6.01±0.35, 3.64±0.15, 2.89±0.18 vs. 6.34±0.23, p-IκB-α/IκB-α: 2.27±0.13, 2.14±0.22, 1.52±0.14 vs. 2.96±0.20, SOD (kU/L): 195.7±19.3, 207.1±25.6, 225.8±23.1 vs. 188.2±26.6, all P < 0.05]. There was no significant effect on lung by 200 mg/kg thalidomide alone. Conclusion Thalidomide has a protective effect on ALI induced by PQ poisoning in rats in a dose-dependent manner, the mechanism may be achieved by reducing the level of oxygen free radicals, reducing the inflammatory factor and inhibiting the IκB-α/NF-κB signal pathway activation.
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Objective To explore the effects of bone marrow mesenchymal stem cells (BMSCs) on viral myositis in mice. Methods Four-week-old BALB/C male mice were randomly divided into normal control group, myocarditis group, and BMSCs intervention group at different stages (3 days and 2 weeks). The mouse model of viral myocarditis was established by intraperitoneal injection of Coxsackie virus B3. The mice in the intervention group were injected with BMSCs in the tail vein at 3 days and 2nd week after the injection of the virus. Four weeks later, echocardiography was performed, and the pathological integral and collagen volume fraction (CVF) were observed and calculated by light microscopy. The qRT-PCR method was used to detect the mRNA expression of homogenates collagen I (col1α1) and collagen fiber III (col3α1) in myocardial tissue. Results Compared with the normal control group, the left anterior and posterior wall became thinner, the diameter and volume of the left ventricle at end systolic period was increased; left ventricular ejection fraction (LVEF) and short axis shortening rate (FS) decreased in the myocarditis group. The differences were statistically significant (P all<0.05). The LVEF and FS in each subgroup of the intervention group were better than those of the myocarditis group, and the improvement in the intervention group was more obvious at the 2nd week after the treatment of the myocarditis. The differences were significant (P all<0.05). Light microscope showed that myocardial CVF in myocarditis group was higher than in normal control group, and CVF in intervention group was reduced compared with myocarditis group and CVF in the 2nd week intervention group was lower than that in the 3 day intervention group. The differences were significant (P all<0.05). Compared with the control group, the mRNA expressions of col1α1 and col3α1 in the myocarditis group were increased, and they were lower in the intervention group than in the myocarditis group, and the differences were significant (P all<0.05). Conclusions BMSCs can reduce the degree of cardiac fibrosis and improve cardiac function in mice with viral myositis, and the intervention effect is better when the virus is infected in the 2nd week.
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We preliminarily investigated the role of NLRP3 inflammasome in Helicobacter pylori (H.pylori) Tipa-induced pro-inflammatory cytokines secretion in PMA-differentiated human acute monocytic leukemia cell line THP-1.PMA-differentiated THP-1 cells were treated with pure recombinant Tips protein.The secretion levels of TNF-α,IL-1β and IL-18 in supernatant of culture medium were detected by ELISA.Then we blocked MyD88/NF-κB and NLRP3/Caspase-1 pathways by PDTC or the general ROS scavenger,NAC,respectively,and determined the secretion levels of proinflammatory cytokines and the expression levels of NLRP3 and Caspase-1.The results showed that Tips protein can significantly induced the secretion of TNF-α,IL-1β and IL-18 in THP-1 cells in a time and dose-dependent manner.Levels of TNF-α,IL-1β and IL-18 approached their peaks at 6 h post-treatment by 40 μg/mL of Tipα protein (P＜0.05).Moreover,the blockade of NF-κB signaling pathway by PDTC can inhibit the secretion of proinflammatory cytokines and the expression of NLRP3 and Caspase-1.When THP-1 cells were pre-treated with ROS scavenger NAC,the Tipα-induced increased IL-1β and IL-18 secretion was obviously eliminated (P＜0.05),while TNF-α level had no significant difference,and the expression levels of Caspase-1 and NLRP3 also have a significant decrease.Our results demonstrated that Tipα can promote THP-1-drived macrophages to secrete proinflammatory cytokines TNF-α,IL-1β and IL-18,and the NLRP3/Caspase-1 pathway may be involved in the Tipα protein-induced IL-1β and IL-18 secretion.