OBJECTIVE: To evaluate the dynamic changes of glucocorticoid (GC) dose and the feasibility of GC discontinuation in rheumatoid arthritis (RA) patients under the background of Chinese medicine (CM). METHODS: This multicenter retrospective cohort study included 1,196 RA patients enrolled in the China Rheumatoid Arthritis Registry of Patients with Chinese Medicine (CERTAIN) from September 1, 2019 to December 4, 2023, who initiated GC therapy. Participants were divided into the Western medicine (WM) and integrative medicine (IM, combination of CM and WM) groups based on medication regimen. Follow-up was performed at least every 3 months to assess dynamic changes in GC dose. Changes in GC dose were analyzed by generalized estimator equation, the probability of GC discontinuation was assessed using Kaplan-Meier curve, and predictors of GC discontinuation were analyzed by Cox regression. Patients with <12 months of follow-up were excluded for the sensitivity analysis. RESULTS: Among 1,196 patients (85.4% female; median age 56.4 years), 880 (73.6%) received IM. Over a median 12-month follow-up, 34.3% (410 cases) discontinued GC, with significantly higher rates in the IM group (40.8% vs. 16.1% in WM; P<0.05). GC dose declined progressively, with IM patients demonstrating faster reductions (median 3.75 mg vs. 5.00 mg in WM at 12 months; P<0.05). Multivariate Cox analysis identified age <60 years [P<0.001, hazard ratios (HR)=2.142, 95% confidence interval (CI): 1.523-3.012], IM therapy (P=0.001, HR=2.175, 95% CI: 1.369-3.456), baseline GC dose ⩽7.5 mg (P=0.003, HR=1.637, 95% CI: 1.177-2.275), and absence of non-steroidal anti-inflammatory drugs use (P=0.001, HR=2.546, 95% CI: 1.432-4.527) as significant predictors of GC discontinuation. Sensitivity analysis (545 cases) confirmed these findings. CONCLUSIONS RA patients receiving CM face difficulties in following guideline-recommended GC discontinuation protocols. IM can promote GC discontinuation and is a promising strategy to reduce GC dependency in RA management. (Trial registration: ClinicalTrials.gov, No. NCT05219214).
Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Arthritis, Rheumatoid/drug therapy* ; Glucocorticoids/therapeutic use* ; Medicine, Chinese Traditional ; Retrospective Studies

OBJECTIVE: To reveal the effects and potential mechanisms by which synaptic vesicle glycoprotein 2A (SV2A) influences the distribution of amyloid precursor protein (APP) in the trans-Golgi network (TGN), endolysosomal system, and cell membranes and to reveal the effects of SV2A on APP amyloid degradation. METHODS: Colocalization analysis of APP with specific tagged proteins in the TGN, ensolysosomal system, and cell membrane was performed to explore the effects of SV2A on the intracellular transport of APP. APP, β-site amyloid precursor protein cleaving enzyme 1 (BACE1) expressions, and APP cleavage products levels were investigated to observe the effects of SV2A on APP amyloidogenic processing. RESULTS: APP localization was reduced in the TGN, early endosomes, late endosomes, and lysosomes, whereas it was increased in the recycling endosomes and cell membrane of SV2A-overexpressed neurons. Moreover, Arl5b (ADP-ribosylation factor 5b), a protein responsible for transporting APP from the TGN to early endosomes, was upregulated by SV2A. SV2A overexpression also decreased APP transport from the cell membrane to early endosomes by downregulating APP endocytosis. In addition, products of APP amyloid degradation, including sAPPβ, Aβ _1-42, and Aβ _1-40, were decreased in SV2A-overexpressed cells. CONCLUSION These results demonstrated that SV2A promotes APP transport from the TGN to early endosomes by upregulating Arl5b and promoting APP transport from early endosomes to recycling endosomes-cell membrane pathway, which slows APP amyloid degradation.
Amyloid beta-Protein Precursor/genetics* ; Membrane Glycoproteins/genetics* ; Animals ; Protein Transport ; Nerve Tissue Proteins/genetics* ; Humans ; Mice ; Endosomes/metabolism* ; trans-Golgi Network/metabolism*

ObjectiveTo construct a cough model induced by chemical stimuli by whole-body plethysmography （WBP） for counting coughs based on cough waveforms， and use this model to explore the antitussive effect of GK-A. MethodDifferent chemical stimuli were used to induce coughs in mice or guinea pigs. Respiratory waveforms were monitored by WBP， and the recognizable and typical cough waveforms were selected for cough counting. Guinea pigs were induced to cough with different concentrations of citric acid or capsaicin， and cough waveforms were used to optimize the stimulation conditions. The optimized guinea pig model of cough was validated with dextromethorphan， and the optimized guinea pig model of capsaicin-induced cough was used to evaluate the antitussive effect of GK-A. ResultWBP could count the coughs induced by capsaicin and citric acid in guinea pigs by recognizable and typical respiratory waveforms. The optimized stimulation conditions were capsaicin concentration of 100 µmol·L^-1 and nebulization for 2 min. The validation results showed that compared with the model group， the dextromethorphan group of guinea pigs had reduced coughs （P<0.05） and prolonged cough latency （P<0.01）. GK-A prolonged the cough latency （P<0.05） and reduced coughs （P<0.05） in the mouse model of ammonia-induced cough. In the guinea pig model of capsaicin-induced cough， GK-A prolonged cough latency （P<0.05）， reduced coughs （P<0.05）， and decreased substance P （SP） content in the guinea pig serum （P<0.05， P<0.01）. ConclusionA guinea pig model of capsaicin-induced cough was successfully established based on cough waveform counting， which provided an objective and accurate cough counting method. GK-A has antitussive effects， possibly by inhibiting the neuropeptide SP.

ObjectiveTo study the effect of Jinlida granules on visceral fat accumulation and its induced inflammatory response in prediabetic rats. MethodMale SD rats were randomly divided into normal group， model group， Jinlida low-dose group （1.5 g·kg^-1）， Jinlida high-dose group （3.0 g·kg^-1） and atorvastatin group （10 mg·kg^-1）. Prediabetic rat model was established using high-carbohydrate， high-fat diet combined with low-dose streptozotocin （STZ） by multiple small-dose intraperitoneal injections. After 8 weeks of modeling and drug intervention for 13 consecutive weeks， body weight， oral glucose tolerance test（OGTT）， fasting blood glucose （FBG）， fasting insulin （FINS）， insulin resistance index （HOMA-IR）， total cholesterol （TC）， low-density lipoprotein cholesterol （LDL-C） and high-density lipoprotein cholesterol （HDL-C） were measured in each group of rats. The content of visceral fat was quantified by micro-computed tomography （Micro-CT）. Hematoxylin-eosin staining （HE） was used to observe the pathological changes of fat cells. The levels of tumor necrosis factor-α （TNF-α） and interleukin- 6 （IL-6） in rat visceral fat and serum were determined by enzyme linked immunosorbent assay （ELISA）. The expression of macrophage marker CD68 in visceral fat was detected by immunofluorescence and Western blot. ResultCompared with normal group， model group had increased oral glucose tolerance， FBG， FINS， HOMA-IR， TC， LDL-C （P<0.01）， elevated body weight and visceral fat accumulation （P<0.05， P<0.01）， enhanced CD68 protein expression and TNF-α and IL-6 levels （P<0.01）， decreased HDL-C （P<0.01）， and abnormal hypertrophy of adipocytes. Compared with model group， Jinlida high- and low-dose groups lowered oral glucose tolerance， HOMA-IR， TC and LDL-C （P<0.05， P<0.01）， body weight and visceral fat accumulation （P<0.05）， and CD68 protein expression and TNF-α and IL-6 levels （P<0.05， P<0.01） and lessened hypertrophy of fat cells. ConclusionJinlida can improve the insulin resistance in prediabetic rats by reducing visceral fat accumulation and its induced inflammatory response， which provides a new pharmacological basis for clinical treatment of prediabetes by Jinlida granules.

Aim Human TMPRSS2 is a transmembrane serine protease.In this paper, the structure and func¬tion of the protein were systematically analyzed by bioinformatics, the codon was optimized and the pro- karvotie expression vector was constructed to explore the molecular mechanism of SARS-CoV-2 infecting host cells.Methods The recombinant expression vector pET-22b-TMPRSS2 was generated by molecular clo¬ning technology.The homology, functional sites, sub¬cellular localization, three-dimensional structure and evolutionary characteristics of TMPRSS2 protein were systematically analyzed by using analytical tools such as Protparam, NetPhos3.1, Blast, Clustal X2 and MEGA7.0.Results The prokarvotic expression plas- mid was constructed correctly; TMPRSS2 belongs to medium molecular weight protein, which is composed of 492 amino acid residues.The theoretical isoelectric point is 8.12, the molecular extinction coefficient is 118 145 L • mol~1 • cm"1 , and the half-life is 30 h; TMPRSS2 has 15 potential glycosylation sites and 49 possible phosphorylation sites.It is a transmembrane hydrophilie protein without signal sequenee.In addi¬tion, the protein has 13 potential B-cell epitopes and 7 T-eell epitopes.Seeondarv structure analysis showed that random coil accounted for the highest proportion of TMPRSS2 protein ( 0.453 3) , followed by extended strand (0.252 0).Sequence comparison and evolu¬tionary analysis showed that the highest sequence con¬sistency and closest genetic relationship with human TMPRSS2 was Pan troglodytes, followed by gorilla.Conclusions Human-derived TMPRSS2 protein is ev- olutionarilv conserved and functionally important.Hie results of this study can help to reveal the structure and mechanism of action of TMPRSS2 protein, provide ide¬as for the diagnosis and treatment of COYID-19, and accelerate the research and development process of new drugs targeting TMPRSS2 protein.

Objective To study the relationship among rs505802 in SLC22A12,rs6855911,rs737267,rs12498742, rs7442295, rs734553, rs16890979 in SLC2A9 genetic polymorphisms and hypouricemia in Ningxia.Methods 6 056 subjects were collected by multistage,stratified random cluster sampling method in October and November in 2011 in Ningxia Hui autonomous region, 98 subjects with hypouricemia were selected.According to gender and age,84 controls were selected.Physical examination and laboratory biochemical index test were conducted for the study population.T test was used to compare general clinical data and biochemical indexs between two groups. SNPs were detected by Sequenom Mass ARRAY technology.By x2test,we compared the frequencies of the geno-type and allele in each group.Samples representativeness was confirmed through the Hardy-Weinberg inspection. Results The levels of TC, LDLC, and Cr in the patients were lower than those in the control group(P<0.05). There were significant differences in the distribution of A,G allele frequencies of SLC2A9 gene rs7442295 between two groups.The risk of hypouricemia in patients with A/A genotype was lower than that of A/G genotype(Pc<0.05),indicating that A>G mutation was associated with hypouricemia.Conclusions Polymorphisms of SLC2A9 gene rs7442295 are significantly correlated with hyporuricemia in Ningxia.

Objective To dynamically observe the early change of thyroglobulin(Tg) levels after (131)I therapy in differentiated thyroid cancer(DTC) patients. Methods The study enrolled 22 post-total-thyroidectomy DTC patients and they were stratified as low to intermediate recurrence according to the 2009 American Thyroid Association Guidelines. The clinical data including pre-ablation stimulated Tg (ps-Tg),corresponding thyroid stimulating hormone(TSH),anti-thyroglobulin (TgAb) values,and the afterwards parameters were dynamically measured each week in the first month after (131)I therapy. Values collected at the first time were defined as Tg 0 and TSH0,while Tg1 and TSH1 were collected at the first week after (131)I therapy respectively. Then the variation trend curves of Tg were drawn,and factors influencing the variation of Tg were analyzed. Two groups were divided according to Tg levels:G1 (Tg≤0.1 ng/ml,n=9) and G2(Tg>0.1 ng/ml,n=13). Results The rates of negative Tg were 4.5%,18.0%,27.0%,36.0%,and 41.0%,respectively,exactly before (131)I therapy and the 1(st),2(nd),3(rd),and 4(th) week after the therapy. One-way analysis of variance showed that the two groups statistically differed in age (F=3.182,P=0.04) and remnant thyroid (U=4.849,P=0.026). Multivariate logistic regression analysis showed that early negative Tg was related to remnant thyroid tissue (OR:2.132;95%Cl:1.418- 6.532,P=0.009). Conclusions Negative Tg can be achieved in nearly half of DTC patients by the end of first month after (131)I therapy. The negative conversion is closely related with the volume of remnant thyroid tissue.
Autoantibodies ; blood ; Humans ; Iodine Radioisotopes ; therapeutic use ; Neoplasm Recurrence, Local ; Thyroglobulin ; blood ; Thyroid Neoplasms ; radiotherapy ; Thyroidectomy ; Thyrotropin ; blood

To research the pharmacokinetic of Danshensu in brain via microdialysis method and automated blood technique. A microdialysis probe was inserted into the left lateral ventricle, and then dialysate samples and blood samples were continuously collected after iv Danshensu. LC-MS/MS was used to determinate for Danshensu in the dialysate samples. The in vivo recovery was used for the calibration of probe. WinNonlin was used for analyzing all pharmacokinetic data. Pharmacokinetic parameters of DSS in blood and in brain showed that Ke, t1/2,, AUC0-t, MRT were 0.04, 0.018 min(-1), 16.64, 58.76 min, 812.59, 51.19 min x mg x L(-1), 15.28, 79.97 min, respectively. The results were indicated that the study was successfully established LC-MS/MS detection method for Danshensu. Microdialysis combined with automated blood technique could better reflect the dynamic characteristics of Danshensu in the rat brain, and it provides a new perspective for pharmacokinetic study.
Animals ; Brain ; metabolism ; Brain Chemistry ; Lactates ; blood ; pharmacokinetics ; Male ; Microdialysis ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo analyze the clinical efficacy of imatinib mesylate (IM) for Ph-positive or BCR-ABL positive chronic myeloid leukemia (CML) to couple the trough plasma concentrations (C mins) of IM with clinical responses and adverse events (AEs).

METHODSOne hundred and one CML patients received IM therapy, and Cmins of IM were determined in 30 patients.

RESULTS(1) Cumulative complete hematological response (CHR), major cytogenetic response (MCyR), complete cytogenetic response (CCyR) and negative BCR/ABL fusion gene rates were 96.6%, 86.5%, 77.5% and 47.2%, respectively, in CML-CP patients. In accelerated and blastic phases (AP and BC) patients, CHR, MCyR, CCyR and negative BCR-ABL fusion gene rates were 58.3%, 25.0%, 25.0%, 8.3%, respectively. (2) Mean Cmins of IM was significantly higher in the CCyR at 1 year [(1472 +/- 482) microg/L] group than in the non-CCyR at 1 years group [(1067 +/- 373) microg/L] (P < 0.05), and higher in the MMR at 1 year group than in the non-MMR at 1 years group [(1624 +/- 468) microg/L vs (1137 +/- 404) microg/L, P < 0.05].

CONCLUSIONIM significantly improves cytogenetic and molecular response, event-free survival, and overall survival for patients with Ph-positive CML. The Cmins of IM exerts a significant impact on clinical response (CCyR and MMR at 1 year).

Adolescent ; Adult ; Aged ; Antineoplastic Agents ; blood ; therapeutic use ; Benzamides ; Female ; Humans ; Imatinib Mesylate ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; blood ; drug therapy ; Male ; Middle Aged ; Piperazines ; blood ; therapeutic use ; Pyrimidines ; blood ; therapeutic use ; Treatment Outcome ; Young Adult

BACKGROUNDMesenchymal stem cells (MSCs) transplantation is of therapeutic potential after ischemic injury in both experimental and clinical studies. Clinically, elderly patients are more vulnerable to acute myocardial infarction (AMI). But little is known about the characteristics of young donor-derived MSCs transplanted to old patients with AMI. The present study was designed to investigate the effect of transplanted MSCs from rats of different ages on the improvement of heart function after AMI.

METHODSMSCs from Sprague-Dawley (SD) rats were isolated and cultured in vitro. The apoptosis characteristics of MSCs were observed under conditions of ischemia and anoxia. SD rats underwent MI received intramyocardial injection of MSCs from young donor rats (n = 8), old donor rats (n = 8), respectively. AMI control group received equal volume physiological saline. Immunofluorescence was used to observe the differentiation of the grafted cells into cardiomyocytes. Four weeks after cell transplantation, reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry for vascular endothelial growth factor (VEGF), VIII-factor immunohistochemistry for vessel density, TUNEL, caspase-3 for cardiomyocyte apoptosis, echocardiography and hemodynamic detection for heart function were performed.

RESULTSThe apoptosis rate of the old donor-derived MSCs group was significantly higher than that of the young donor-derived MSCs group under conditions of ischemia and anoxia (P < 0.05). Engrafted MSCs survived, proliferated and differentiated into myocardium-like cells. VEGF gene expression and capillary density in the old donor-derived group were lower than those in the young donor-derived group but higher than those in the control group (P < 0.05). The transplantation of old donor-derived MSCs attenuated apoptosis of cardiomyocytes in the peri-infarct region compared with the control group and the effect was elevated in young donor-derived MSCs (P < 0.05). The heart functions (left ventricle ejection fraction (LVEF), left ventricle fractional shortening (LVFS)) were improved more significantly in the old donor-derived MSCs group than in the control group and the heart function in the young donor-derived MSCs group further improved (P < 0.05).

CONCLUSIONSYoung donor-derived MSCs can improve heart function significantly through angiogenesis and decreasing cardiomyocyte apoptosis when transplanted to the infarcted area.

Age Factors ; Animals ; Apoptosis ; Caspase 3 ; metabolism ; Cells, Cultured ; Flow Cytometry ; Immunohistochemistry ; In Situ Nick-End Labeling ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Mesenchymal Stromal Cells ; cytology ; Myocardial Infarction ; physiopathology ; surgery ; Myocytes, Cardiac ; cytology ; enzymology ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Vascular Endothelial Growth Factor A ; genetics ; metabolism