The traditional theory of "the spleen governs the dispersion of essence" refers to the spleen's pivotal role in distributing refined nutrients throughout the body. In traditional Chinese medicine （TCM）， lipids are categorized under "gaozhi （膏脂）"， and their transportation and metabolism via apolipoproteins are believed to be closely related to the spleen's dispersing function. The liver， which synthesizes apolipoproteins， is functionally linked to the spleen system in TCM. Impaired dispersion of essence by the spleen and disrupted transportation of gaozhi constitute the pathological mechanism of dyslipidemia due to spleen deficiency. Strengthening the spleen and resolving dampness is the core therapeutic principle. From the perspective of modern medicine， this may involve promoting hepatic functional recovery related to lipid metabolism， thereby enhancing lipid processing and reducing the levels of abnormally accumulated lipids in the bloodstream.

Objective To develop and optimize the extraction method of exosomal miRNA and compare it with traditional methods. Methods The exosomal miRNA of MSC,NK and CIK cells was used as the research subject. The removal efficiency of genomic DNA from exosomal miRNA by gDNA removal column was detected by UV spectrophotometer and agarose gel electrophoresis. The lysates(exosome G2 lysate and exosome miRNA lysate)and aggregation reagents(absolute ethanol and isopropanol)were optimized by using concentration,purity and gene expression level(Ct value)as evaluation indexes.Exosomal miRNA of MSC,NK,CIK cells and healthy human serum was extracted by the developed method,Trizol method and Trizol magnetic beads method,and detected by RT-qPCR. Results The gDNA removal column effectively removed genomic DNA from exosomal miRNA. The concentrations of exosomal miRNA extracted from MSC,NK and CIK cells by using exosome miRNA lysate were significantly higher than those by using exosome G2 lysate(t = 6. 358,P = 0. 020). The purity of miRNA samples extracted by exosome G2 lysate was low,and there was foreign protein pollution,but exosome miRNA lysate effectively removed the pollution. The Ct values of miR-Let-7i,miR-16-1 and miR-150 genes in exosomal miRNA of CIK cells extracted by exosome miRNA lysate were significantly lower than those by exosome G2 lysate(t = 30. 120,P =0. 008). The concentration of exosomal miRNA extracted from MSC,NK and CIK cells by isopropanol was significantly higher than that by absolute ethanol(t = 8. 567,P = 0. 010),and the purity was significantly lower than that by absolute ethanol(t = 6. 214,P = 0. 020). The Ct values of miR-Let-7i,miR-16-1 and miR-150 genes in exosomal miRNA extracted from CIK cells by two aggregation reagents had no significant difference(t = 2. 297,P = 0. 120). Compared with Trizol method and Trizol magnetic beads method,the expression of miR-Let-7i,miR-16-1,miR-Let-7a and miR-150 genes in exosomal miRNA extracted from CIK,NK,MSC cells and healthy human serum by the developed method was more abundant,and the overall Ct value was lower. The dissolution peak was prominent and sharp,exhibiting a single main peak. Conclusion The exosomal miRNA extracted by the developed method has high concentration and purity with stable Ct value,and the method has high sensitivity and good specificity. This study lays a foundation for further research on exosomal miRNA.

AIM: To investigate the mechanism of elemene synergistic bortezomib against multiple myeloma based on ROS-NF-κB-p38MAPK signaling pathway. METHODS: CCK-8 assay was used to detect cell activity. Nude mice were randomly divided into control group, bortezomib (BTZ) group, elemene (ELE) group and combination group. Each group was treated with BTZ, ELE and BTZ combined with ELE, respectively. Tunel staining was performed to observe the apoptosis of tumor tissues. The expressions of Caspase-3, Bcl-2, NF-κB and p38 MAPK were detected by Western Blot. Cell cycle, apoptosis and reactive oxygen species (ROS) expression were detected by flow cytometry using human myeloma U266 cells. RESULTS: When 4.0 μmol/L ELE combined with 50 nmol/L BTZ treated U266, the cell activity was significantly reduced compared with that of NC, BTZ and ELE groups (P< 0.05). The tumor volume of nude mice in BTZ group, ELE group and combined group was significantly reduced compared with the control group (P <0.05), and the combined group was the smallest. Tunel staining results showed that the apoptosis level in the control group was lower than that in the BTZ group, ELE group and the combined group (P<0.05), and the combined group had the lowest apoptosis level. Compared with the control group, the expressions of Caspase-3 and p38 MAPK in BTZ group, ELE group and combination group were significantly increased, while the expression of Bcl-2 was significantly decreased. The apoptosis level and expression of ROS in BTZ group, ELE group and the combined group was significantly increased compared with the control group (P<0.05). CONCLUSION: ELE can enhance the role of BTZ in promoting apoptosis of myeloma cells, which may be achieved by regulating ROS/NF-κB/p38 MAPK signaling pathway to enhance the level of apoptosis of tumor cells to achieve anti-tumor effect.

Objective:To explore the method of constructing an early mortality risk prediction model for patients with sepsis-induced cardiorenal syndrome by machine learning algorithm, so as to provide a basis for early clinical identification of high-risk patients and accurate treatment.Methods:Patients with sepsis-induced cardiorenal syndrome from January 1, 2015 to May 31, 2019 in Tongji Hospital, Tongji University were enrolled. Basic characteristics, laboratory indexes, hospitality treatment and other relevant baseline data were collected. Thirty-day mortality was defined as the primary end-point event after the enrolled patients were diagnosed. Python software was applied to establish different machine learning models, and the area under the receiver -operating characteristic curve ( AUC) was used to evaluate the predictive value of models. Disease-related risk factors were selected according to the most optimal model. Importantly, visualized decision tree and semi-naive Bayesian (sNB) models were established to further explore the interrelationship between these risk factors. Results:A total of 340 patients were included, of whom 114 patients (33.5%) died within 30 days after diagnosis. The AUC of support vector machine (SVM), random forest (RF), gradient boosting decision tree (GBDT), extreme gradient boosting (XGBoost), and light gradient boosting machine (LGBM) prediction models were 0.652, 0.868, 0.870, 0.754, and 0.852, respectively. The AUC of GBDT model had the most efficiency to predict end-point events, and the prediction AUC value was better. According to the feature ranking of GBDT model, the relevant influencing factors were selected, including total sequential organ failure assessment (SOFA) score, neural SOFA score, vasoactive drug application, cardiac troponin I (cTNI), age, myoglobin, circulation system SOFA score, chronic kidney disease, heart rate and baseline serum creatinine. Visualized decision tree model had 4 layers, 15 nodes and 8 terminal nodes as evidenced by total SOFA score, myoglobin, baseline serum creatinine and age. The total SOFA score, change rate of myoglobin, serum creatinine and age were included into the visualized decision model. The AUC value of the model for predicting end-point event was 0.690. sNB model revealed complex correlation between the risk factors, in which neural SOFA score was related to total SOFA score, vasoactive drug application was related to total SOFA score, and cTNI was related to baseline serum creatinine. Conclusions:A risk prediction model for patients with sepsis-induced cardiorenal syndrome is established and the model showes that high SOFA score remains the primary risk factor for patients with sepsis-induced cardiorenal syndrome based machine learning. Visualized decision tree and sNB models help clinicians to further identify the dependence and logic relationship among these risk factors clearly and provide a novel method to predict mortality risk for patients with sepsis-induced cardiorenal syndrome.

Animals ; Fasting ; metabolism ; Membrane Proteins ; metabolism ; Mice ; Sterol Regulatory Element Binding Protein 1 ; metabolism ; Transcription Factors ; deficiency ; metabolism

Objective This study was performed to investigate the levels of HSP70 in tissue in pemphigus as a possible new theoretical basis for further elucidate the pathogenesis of pemphigus.Methods The expression of HSP70 in 62 patients with pemphigus was determined by immunohistochemistry,and the normal skin was taken as control.Results The results showed that the positive cells of HSP70＞75 ％ in the blisters of pemphigus vulgaris and the positive cells of HSP70＞50％ in the inflammatory cells near the blisters,and the expression of HSP70 was significantly higher than that in normal skin,which was statistically significant(Z=5.42,4.73,P＜0.01).Conclusion The abnormal expression of HSP70 in inflammatory cells and psoriasis of pemphigus patients showed that HSP70 is involved in the pemphigus.

Intervertebral disc degeneration (IDD) is one of major causes for intervertebral disc degenerative diseases, and patients with IDD usually suffer from serious low back pain. The current treatments for patients with IDD only relieve the clinical symptom rather than restore biological balance of IDD, leading to inadequate and unsatisfactory results. MicroRNAs (miRNAs) are endogenous, non-coding, single-stranded RNA molecules, which regulate the gene expression at the post-transcription levels. Research evidences support the involvement of miRNAs in many biological processes, such as lipid metabolism, apoptosis, differentiation and organ development. Accumulating evidences indicate that the expressions of miRNAs change significantly in degenerative tissues. In addition, dysregulated miRNAs contribute to multiple pathological process of IDD, including proliferation and apoptosis of nucleus pulposus and extracellular matrix components, inflammatory response and cartilage endplates degeneration. In this review article, we summarize the expression profiles and roles of miRNAs in IDD, which may provide a novel strategy of biological therapy for the disease.
Apoptosis ; Extracellular Matrix ; pathology ; Gene Expression ; Gene Expression Profiling ; Humans ; Intervertebral Disc Degeneration ; genetics ; pathology ; MicroRNAs ; genetics

Objective:To evaluate immune response of murine peritoneal macrophage challenging by methicillin-resistant S.aureus(MRSA)after pretreatment with Pam3Csk4(TLR2 agonist).Methods: Murine peritoneal macrophage was pretreated with Pam3Csk4(1 μg/ml).Following pretreatment 12 h later,heat-killed MRSA( HK-MRSA) was added and incubated for another 2 or 6 hours.The protein and mRNA level of TNF-α, IL-6 and IL-1 were determined by ELISA and Q-PCR, respectively.To estimate phagocytosis of macrophage,HK-MRSA/MSSA labeled with FITC( FITC-HK-MRSA/MSSA) were added to well and incubated for 30 min.After washing 5 times with PBS,intracellular FITC-HK-MRSA was detected by flow cytometry.To estimate antimicrobal activity of macrophage,live MRSA and MSSA were added to well and incubated at indication time,the CFU of s.aureus was estimated via a 10-fold serial dilution on agar media.cDNA was further quantitative assessed using primers for mouse FCR-Ⅰ,FCR-Ⅲ,CR-1,CR-3,iNOS and LL37 by Q-PCR .Results: Compared with saline-pretreated cell, the protein and mRNA level of TNF-α, IL-6 and IL-1 were markely reduced, respectively.However, both the phagocytosis and antimicrobal activity to S.aureus were significantly increased in macrophages pretreated with Pam3Csk4.Further study found that the macrophages had higher FCR-Ⅰ,FCR-Ⅲ,CR-1,CR-3,iNOS and LL37 expression at 6 h and 12 h post-stimulation Pam3Csk4.Conclusion: The results suggest that Pam3Csk4 could activate murine antimicrobal activity of peritoneal macrophage challenging by methicillin-resistant Saureus via increasing opsonophagocytosis in depended antibodies, complements manners.The results suggest Pam3Csk4 probably be a novel immunotherapy candidate against MRSA.

Objective To Analyze CD4+ cell count which embodies curative effect in HIV and patients with AIDS, who have treated with TCM for half a year. Methods According to CD4+cell count, the patients were divided into 4 phases. Their CD4+cell count were analyzed before and after the treatment. Results 1.Rank sun test showed that CD4+cell count were significantly improved in people who used TCM treatment. On the whole, CD4+cell count was(317.76±175.61) in 1 cu mm before treatment, which was(350.60±175.92) in 1 cu mm after treatment, P<0.01. 2. Ridit showed that patients whose CD4+cell count more than 500 were in phase I. Their R=0.614, and the 95%confidence interval was 0.5702 to 0.6579. Patients whose CD4+cell count more than 350 and less than 500 were in phase II. Their R=0.575, and the 95%confidence interval was 0.5439 to 0.6062. Patients whose CD4+cell count more than 200 and less than 350 were inphase III. Their R=0.460 and the 95%confidence interval was 0.4347 to 0.4849. Patients whose CD4+cell count less than 200 were in phase IV. Their R=0.428, and the 95%confidence interval was 0.3971 to 0.4589. There was no overlap between phase III, phase IV and phase I, phase II in 95% confidence interval. Conclusion TCM has the advantages in strengthening vital qi, but that is worse than western medicine in effort of expelling pathogen.

Objective Determine the common syndrome elements of viral hepatitis.Methods Refering to the syndrome elements hypothesis proposed by Zhang Zhibin,syndrome elements from the collection of 19,341 cases of Viral Hepatitis were extracted which were diagnosed by National Science and Technology Major Project of AIDS and Viral Hepatitis and Other Major Infectious Diseases Prevention and Control.And the results were compared with industry standards.Results There were 27 involved syndrome elements of viral hepatitis.Common syndrome elements were Qi stagnation,internal dampness,internal fire,blood stasis,yin deficiency,Qi deficiency,toxin,pathogenic water,and yang deficiency; The main excessive syndromes were Qi stagnation,internal dampness,internal fire,and blood stasis.The main deficiency syndromes were yin deficiency and Qi deficiency.Conclusion Excessive syndromes were the main syndromes of viral hepatitis.Dampness,heat,blood stasis,and toxin played key roles in the development of viral hepatitis.In addition to this,Qi stagnation,pathogenic water and phlegm were also important syndrome elements.