BACKGROUND:Spleen has the functions of blood storage,hematopoiesis,and immunity.With the increase of age,the structural degeneration and functional decline of spleen lead to the impairment of immune system function,thus accelerating the aging process of the body.The treatment of spleen aging in tree shrews with highly active umbilical cord mesenchymal stem cells has not been reported. OBJECTIVE:To explore the intervention effect and mechanism of highly active umbilical cord mesenchymal stem cells on spleen aging in tree shrews. METHODS:Highly active umbilical cord mesenchymal stem cells were isolated,cultured,and obtained from the umbilical cord tissue of newborn tree shrews by caesarean section.The differentiation abilities of adipogenesis,osteogenesis,and chondrogenesis were detected by three-line differentiation kit.Cell cycle and surface markers were detected by flow cytometry.The second generation of highly active umbilical cord mesenchymal stem cells were transfected with Genechem Green Fluorescent Protein with infection complex values of 100,120,140,160,180,and 200,respectively,to screen the best transfection conditions.After transfection,the fourth generation of highly active umbilical cord mesenchymal stem cells was injected into the tail vein of tree shrews in the elderly treatment group.The young control group and the aged model group were not given special treatment.After 4 months of treatment,the spleen tissue was taken and the structure of the spleen was observed by hematoxylin-eosin staining.β-Galactosidase staining was used to detect the activity of aging-related galactosidase.Immunohistochemical staining was used to detect the expression levels of p21 and p53 proteins.Ki67 and PCNA immunofluorescence staining was used to detect cell proliferation activity.Immunofluorescence staining was used to detect the expression levels of spleen autophagy protein molecules Beclin 1 and APG5L/ATG5.Reactive oxygen species fluorescence staining was used to detect the content of reactive oxygen species in spleen tissue.CD3 immunofluorescence staining was used to detect the change of the proportion of total T lymphocytes.The secretion levels of interleukin 1β and transforming growth factor β1 in spleen were detected by enzyme linked immunosorbent assay.The distribution of highly active umbilical cord mesenchymal stem cells labeled with green fluorescent protein in spleen tissue was observed by DAPI double staining of nucleus. RESULTS AND CONCLUSION:(1)Highly active umbilical cord mesenchymal stem cells grew in a short spindle shape with fish-like growth,with a large proportion of G0/G1 phase,and had the potential to differentiate into adipogenesis,osteogenesis,and chondrogenesis.(2)Multiplicity of infection=140 and transfection for 72 hours were the best conditions for labeling tree shrews highly active umbilical cord mesenchymal stem cells with Genechem Green Fluorescent Protein.(3)Compared with the aged model group,in the aged treatment group,the spleen tissue cells of tree shrews were arranged closely,and the area of white pulp was increased(P<0.01);the boundary between red pulp and white pulp was clear;the proportion of germinal centers did not show statistically significant difference(P>0.05).The activity level of galactosidase related to spleen tissue aging was decreased(P<0.001),and the expression levels of aging protein molecules p21 and p53 were down-regulated(P<0.001).The expression levels of proliferation-related molecules Ki67 and PCNA were up-regulated(P<0.001,P<0.05);expression levels of autophagy-related molecules Beclin 1 and APG5L/ATG5 were up-regulated(P<0.001),and the content of reactive oxygen species decreased(P<0.001),and the proportion of CD3+T cells increased(P<0.05).The secretion level of interleukin 1β in the aging-related secretion phenotype decreased(P<0.001);no significant difference was found in transforming growth factor β1 level(P>0.05).Compared with the young control group,the above indexes were significantly different in the elderly treatment group(P<0.05).(4)Green fluorescent cells labeled with green fluorescent protein were observed in spleen tissue of tree shrews the elderly treatment group by frozen tissue section observation.The results show that intravenous infusion of highly active umbilical cord mesenchymal stem cells can migrate to spleen tissue,inhibit the production of reactive oxygen species,down-regulate the expression of aging-related proteins,induce autophagy,promote cell proliferation,reduce chronic inflammation,and then improve the structure and function of spleen tissue.

ObjectiveTo analyze the adverse reactions associated with the clinical use of Banxia （Rhizoma Pinelliae）- Wutou （Aconitum） in the same formula， with the aim of providing a reference for the safety of their clinical application. MethodsLiterature on the clinical application of antagonistic herbs "Banxia-Wutou" used in the same formula， published from January 1st， 2014， to June 30th， 2023， was retrieved from databases including CNKI， VIP， Wanfang， SinoMed， PubMed， Cochrane Library， and Embase. A database was established， and information related to adverse reactions was extracted， including descriptions， classifications， specific manifestations， management and outcomes， patients' primary diseases （western medicine diseases and traditional Chinese medicine diagnoses and syndromes）， and medication information （dosage， ratio， administration routes， and dosage forms）. ResultsA total of 79 researches simultaneously used antagonistic herbs Banxia-Wutou in the same formula and reported associated advers reactions. Gastrointestinal adverse reactions were the most common， with 8 studies reporting management of adverse reactions and 3 studies reporting improvement with no intervention. Among the 11 researches， the adverse reaction relieved to extant， while other 69 researches didn't report the managment of adverse reaction and its prognosis. For the primary disease in western medicine system， chronic bronchitis and chronic obstructive pulmonary disease （COPD） were most common， while gastric pain was the most common symptom in traditional Chinese medicine with spleen and kidney deficiency and spleen stomach cold deficiency being the most frequent syndromes. The most common Banxia dosage was 10 g， while for the Wutou， Fuzi （Radix Aconiti Lateralis Praeparata） was predominant with the highest dose at 15 g. The most frequent herbal combination was Banxia-fuzi， with a 1∶1 ratio. The main administration route was oral， and the primary dosage form was decoction. ConclusionGastrointestinal adverse reactions are the most common in the clinical use of Banxia-Wutou antagonistic herb combinations. Research on the safety of "Banxia-Wutou" combinations should focus on respiratory system diseases and spleen-stomach related conditions.

Objective:To compare the prognostic values of different classification by using transpulmonary pressure gradient (TPG), diastolic pressure gradient (DPG) and pulmonary vascular resistance (PVR) in patients with pulmonary hypertension due to left heart disease (PH-LHD), and investigated hemodynamic and clinical factors associated with mortality in patients with PH-LHD.Methods:This was a single-center prospective cohort study. In-hospital patients diagnosed with PH-LHD via right heart catheterization at the Department of Cardiology, the First Affiliated Hospital of Nanjing Medical University, from September 2013 to December 2019 were enrolled. Patients were divided according to TPG (cutoff value 12 mmHg; 1 mmHg=0.133 kPa), DPG (cutoff value 7 mmHg), PVR (cutoff value 3 Wood Units), and the combination of TPG and PVR. Baseline characteristic was recorded. All patients were followed up until the occurrence of endpoint event, defined as all-cause death that occurred during the follow-up period, or until April 18, 2022. Receiver operating characteristic curves were used to compare the predictive value of 3 classification methods for all-cause death in PH-LHD patients. The optimal cutoff values were calculated using Jorden index. Survival analysis was performed using Kaplan-Meier analysis, and log-rank test was used to compare the predictive efficacy of classification methods based on optimal cutoff values or guidance-recommended thresholds for the survival of PH-LHD patients. Variables showing statistical significance in the univariate analysis were incorporated into multivariate Cox regression model to analyze the independent risk factors for all-cause mortality.Results:A total of 243 patients were enrolled, aged (54.9±12.7) years old, including 169 (69.5%) males. During a median follow-up of 57 months, there were 101 (41.6%) deaths occurred. Grouping results were as follows: (1) TPG: TPG≤12 mmHg group 115 patients, TPG>12 mmHg group 128 patients; (2) DPG: DPG<7 mmHg group 193 patients, DPG≥7 mmHg group 50 patients; (3) PVR: PVR≤3 Wood Units group 108 patients, PVR>3 Wood Units group 135 patients; (4) TPG and PVR: TPG≤12 mmHg and PVR≤3 Wood Units group 89 patients, TPG>12 mmHg and PVR>3 Wood Units group 109 patients. PVR ( AUC=0. 698,95% CI:0.631-0.766) had better predictive value for all-cause mortality than TPG ( AUC=0.596, 95% CI: 0.523-0.669) and DPG ( AUC=0.526, 95% CI: 0.452-0.601) (all P<0.05). The optimal cutoff values for TPG, DPG, and PVR were13.9 mmHg, 2.8 mmHg, and 3.8 Wood Units, respectively. Kaplan-Meier analysis based on the optimal cutoff values or guidance-recommended thresholds showed that PVR and TPG were the predictors of survival ( P<0.05), while DPG did not showed significance ( P>0.05). Multivariate Cox regression analysis showed that age, PVR and log 2N-terminal pro-B-type natriuretic peptide were independent risk factors for all-cause mortality in PH-LHD patients (all P<0.05). Conclusion:Classification according to PVR was most valuable in predicting all-cause death in PH-LHD patients, while TPG showed moderate predictive ability and DPG had no predictive value.

Objective:To study the effects of early intervention on physical and neurological development of premature infants.Methods:One hundred and fifty-two premature infants born in Pan′an County Maternal and Child Health Family Planning Service Center from January 2017 to June 2018 were randomly selected. According to parents′ wishes, 75 cases received regular systematic health care and parenting guidance(control group), and 77 cases received targeted intervention guidance and individualized training on the basis of the control group(intervention group). The results of mental development index (MDI) and psychomotor Development index(PDI) were evaluated at 6 and 12 months respectively.Results:Six and 12 months after treatment, the MDI scores of the intervention group were significant higher than those of the control group [(94.2 ± 5.2) scores vs. (85.3 ± 6.1) scores, (104.9 ± 3.9) scores vs. (91.7 ± 5.7) scores], the PDI scores of the intervention group were also significant higher than those of control group [93.7 ± 4.3) scores vs. (81.7 ± 5.5) scores, (96.9 ± 3.8) scores vs. (83.9 ± 4.6) scores], and there were statistical differences ( P<0.05). Results:Early comprehensive intervention for premature infants has a good effect on their intelligence and motor ability development, which is worthy of clinical application.

BACKGROUND: Mesenchymal stem cells attract extensive attention because of good biological characteristics and broad prospects, but the cells gradually show the characteristics of the aging with the increase of individual age or incubation time in vitro. Nonhuman primates have similar biological characteristics with human being, and have unique advantage in the animal model and disease treatment research.OBJECTIVE: To analyze the difference in proliferation and differentiation of bone marrow mesenchymal stem cells from macaques at different ages and to explore the effect of age on bone marrow mesenchymal stem cells and the possible mechanism.METHODS: Bone marrow samples from male macaques aged < 3 years and over 20 years were collected through bone marrow puncture, and divided into young group and elder group, with three macaques in each group. Then, bone marrow mesenchymal stem cells were isolated and cultured in vitro, and the morphological changes, proliferation and differentiation ability were observed. Age-related beta-galactosidase staining was performed, and protein microarray and ELISA were used to detect cytokine levels.RESULTS AND CONCLUSION: With age, the proliferation and differentiation of bone marrow mesenchymal stem cells from the elder macaques were reduced significantly, and the number of senescent cells increased significantly; the levels of interleukin-1b, interleukin-4, interleukin-6, tumor necrosis factor α and vascular endothelial growth factor were elevated obviously, the levels of heparin-binding basic fibroblast growth factor and placental growth factor were reduced. These findings indicate that the body's aging lead to the reduction in the proliferation, differentiation and cytokine secretion of bone marrow mesenchymal stem cells.

Objective To simplify the calculation of T>MIC and evaluate meropenem regimens based on the simple mathematical model of T>MIC reported in literature for two-compartment model.Methods Six meropenem regimens were designed according to the recommended dose with the infusion duration of 0.5 h and 3 h.Four different MIC susceptibility breakpoint of meropenem against clinical microbiologic flora were used to formulate different T>MIC.Each T>MIC was calculated by both the simple and two-compartment model of T>MIC.Meropenem regimens were evaluated for different bacterial infections based on the simple model of T>MIC with 40%~100% of the T>MIC% as the target.The t-test suggested no significant difference between the pairs of T>MIC calculated by the two models.Results and Conclusion Simple model of T>MIC can replace the two-compartment model.Meropenem regimens can be optimized based on this simple model of T>MIC conveniently and quickly.

Objective To investigate the preliminary clinical result of thoracolumbar fracture combined with posterior ligamentous complex injuries repaired by posterior transpedicular screw fixation.Methods A retrospective review was performed on 22 patients with thoracolumbar flexion-distraction fracture combined with posterior ligamentous complex injuries treated with transpedicular screw fixation from July 2008 to March 2013.There were 16 males and 6 females with mean age of 39 years (range,23-62 years).After medically stable,posterior pedicle screw fixation was performed under intravenousinhalational anesthesia.According to the degree of fracture displacement and types of ligament injury,posterolateral bone grafting or intervertebral fusion at the level of injury was conducted.Vertebral height restoration,Cobb' s angle and American Spinal Injury Association (ASIA) score were reviewed preoperatively,at postoperative 3 days and at the last follow-up.Results All the patients were operated on smoothly.There were no complications during operation.All the patients were followed up for 5-51 months (mean,26.5 months).Fracture reductions were satisfied with the closure of vertebral posterior element.Mean anterior vertebral height and Cobb' s angle improved by 20.6％ and 10.60°respectively after operation (P ＜0.01).Eight patients with neurological dysfunction showed some recovery after operation with the mean sensory score improved by 20.7％ (P ＜ 0.05) and mean motor function score improved by 30.9％ (P ＜ 0.0l).All bone grafts were healed,without pain,loosening or breakage in the fixation system.Conclusions Posterior pedicular screw fixation attains good short-term outcome for thoracolumbar flexion-distraction fracture combined with posterior ligamentous complex injuries.The surgery provides satisfactory reduction and instant spinal three-column stability for the unstable spine fracture.Sufficient bone graft is the guarantee to permanent stability.

Objective To investigate the status of Health-related Quality of Life ( HRQOL) in patients with pulmonary hypertension and analyze its influencing factors. Methods A total of 165 patients with pulmonary hypertension were surveyed by General Information Questionnaire, Chinese version of Minnesota Living with Heart Failure-modified for PH ( MLHF-PH ) , Hospital Anxiety and Depression Scale ( HADS ) , Social Support Rating Scale ( SSRS ) , Self-efficacy Management of Chronic Disease ( SECD6 ) and Medical Coping Method Questionnaire (MCMQ). Results The score of MLHF-PH was (41. 93 ± 18. 98), and the scores of MLHF-PH in marital status, different occupations, fee resource and disease type were statistically significant (F =6. 692,8. 593, 3. 342, 4. 388, respectively;P <0. 05). The multiple stepwise regression analysis showed that the severity of mMRC dyspnea, status of depression, 6 minute walk test distance, New York Heart Association ( NYHA ) functional classification, symptoms and times of hospitalization were introduced into the regression model, which could explain 78. 1% of the total variance. Conclusions The general health related quality of life in pulmonary hypertension patients were at a low level. Doctors and nurses should implement particular measurements according to their conditions to improve their quality of life.

In 2009,5 523 participants of She nationality living in Ningde City of Fujian province were multistage-stratified and randomly collected,and 5 080 were enrolled in analysis eventually.The TSH screening (reference range,0.3-3.0 mIU/L) revealed biochemical hyperthyroidism in 2.4％ (2.1％ of males and 2.7％ of females) and biochemical hypothyroidism in 9.7％ (7.4％ of males and 11.4％ of females) of the participants.The prevalence of positive TPOAbs was 10.8％ (7.4％ of males and 13.5％ of females).TSH values outside the normal reference range were associated with a higher prevalence of positive TPOAbs,indicating that autoimmunity seems to be an important factor in both hyperthyroidism and hypothyroidism.

OBJECTIVE: To observe the effect of norcantharidin (NCTD) on the expression of mRNA and protein of fibronectin (FN), collagen IV(Col IV) and transforming growth factor-β1(TGF-β1) in human kidney proximal tubular epithelial (HK)-2 cells induced by high glucose. METHODS: HK-2 cells were incubated with serum-free DMEM for 24 h to synchronize cell growth, and then the cells were divided into 4 groups: Group C (5.5 mmol/L D-glucose), Group M (5.5 mmol/L D-glucose + 24.5 mmol/L-mannitol), Group HG (30 mmol/L D-glucose), and Group HG + NCTD (30 mmol/L D-glucose + 0.5-40 mg/L NCTD). Cytotoxicity of HK-2 cells induced by high glucose of NCTD was detected by Trypan blue dye exclusive assay. The effect of NCTD on the proliferation of HK-2 cells in high glucose was determined by MTT. The cells were collected to extract total RNA and protein at 6, 24 and 48 h after the incubation. The expression of FN, Col IV and TGF-β1 mRNA was examined by RT-PCR, and FN, Col IV and TGF-β1 protein was analyzed by Western blot. RESULTS: Trypan blue dye exclusive assay showed NCTD concentrations over 5 mg/L were rather toxic in HK-2 cells. The proliferation of HK-2 cells in high glucose was interrupted by interfered with 5 mg/L NCTD as measured by MTT (P<0.05). NCTD at 5 mg/L had a stronger inhibitory effect than NCTD at 2.5 mg/L. Real-time PCR and Western blot showed that the mRNA and protein expression of FN, collagen IV and TGF-β1 increased in HK-2 cells treated with high glucose (P<0.05), while that in cells treated by NCTD was dramatically inhibited (P<0.05). No change in these parameters was detected in the 30 mmol/L D-mannitol control group (P>0.05). CONCLUSION NCTD can downregulate FN, collagen IV and TGF-β1 mRNA and protein expression in HK-2 cells stimulated by 30 mmol/L D-glucose.
Bridged Bicyclo Compounds, Heterocyclic ; pharmacology ; Cell Line ; Collagen Type IV ; genetics ; metabolism ; Down-Regulation ; drug effects ; Epithelial Cells ; cytology ; Fibronectins ; genetics ; metabolism ; Glucose ; pharmacology ; Humans ; Kidney Tubules, Proximal ; cytology ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Transforming Growth Factor beta1 ; genetics ; metabolism