Objective To observe the local immue response and changes of angiogenic factors of tumor cells in Heps-bearing mice after mesenchymal stem cell (MSC) are administrated. And to explore the feasibility and safety of MSC for liver tumors therapy. Methods MSC were obtained through adherent culture method. Phe-notypes of MSC were analyzed by flow cytometry. MSC were labeled with DAPI in vitro. 54 Mice of 8 weeks of age with subcutaneously transplanted liver carcinomas were developed randomly. When the maximal diameters of the tumor reached 0.5 - 0.8cm, they were divided into three groups randomly: MSC group, DAPI group and NS control group. 2 × 10~6 MSC and MSC marked by DAPI were administrated into the mice right rear back tumor tissue. The survival time of the tumor-bearing mice was recorded and the mean survival time was calculated. Immunohistochemical staining was performed to count CD4~+ T cells and CD8~+ T cells in the local tumor,as well as to examine the expression of vascular endothelial growth factor ( VEGF) in tumor cells. Results In the MSC group,the mean survival time was 45 d (95%CI;33 ～56 d) ,in the NS control group, the mean survival time was 33 d ( 95%CI : 28 ～ 37 d). There was a statistical significance in the difference between them ( P < 0.05). Immunohistochemical staining results showed as follow: the number of CD4~+ T cells and CD8~+ T cells in the MSC group decreased significantly in comparison with the NS control group at early stage. The expression of VEGF also decreased obviously in comparison with the NS control group and induced tumor cells necrosis at late stage. The survival time of MSC group was prolonged. Conclusion MSC can engraft in Heps-bearing tumor tissue, and inhibit T lymphocyte cellular immunity at early stage. It can reduce the number of CD4~+ T cells and CD8~+ T cells and promote tumor growth. MSC can down regulate VEGF expression and induce tumor cells necrosis at late stage. By this way,it can prolong the survival time of Heps-bearing mice.

Objective:To study the characteristics of intestinal tract pathological injury of rats in hypobaric hypoxia at different high altitude.Methods:30 male SD rats were randomly divided into 5 groups:Plain group (n=6),High-altitude (HA) 5000 m for 10 day group(n=6),HA 5000 m for 21day group(n=6),HA 6500 m for 10day group(n=6),HA 6500 m for 21day group (n=6).Rats were raised normally either in plain or simulated high altitude environment,at the corresponding time point,rats were euthanized,small intestines were harvested,fixed tissues were processed routinely into paraffin and sections were stained routinely with hematoxylin and eosin.Morphologic parameters were measured by optical microscope and then the pathological injury score were evaluated.Results:The pathological injury scores ofjejunnm and ileum in the high altitude group were significantly higher than that of the plain group (P＜0.01),the pathological injury scores of jejunum,ileum and colon were significantly higher in the HA 5000 m for 21 day group than that of the HA 5000 m for 10 day group,but significant lower than that of the HA 6500 m for 21 day group,the pathological injury scores of jejunum,ileum and colon were significantly higher in the HA 6500 m for 10 day group than that of the HA 5000 m for 10 day group(P＜0.01,P＜0.05).The pathological injury scores of colon were significantly higher in the groups of the high altitude than the plain group except for the HA 5000 m for 10 day group (P＜0.01,P＜0.05).The pathological injury scores of jejunum had significant different with that of the colon in the HA 5000 m for 21 day group (P＜0.05);the pathological injury scores of both jejunum and ileum had significant different with that of the colon in the HA 6500 m for 21 day group(P＜0.05,P＜0.01).Conclusions:The injuries of the intestinal mucosa became serious with the with rising altitude and the stayed time,the intestinal injuries were more serious than that of colon at same situation,however there were no significant different between jejunum and ileum,the injuries of colon happened later than intestine,and they correlated with the stay time in the high altitude,which indicated that intestinal injury should be pay more attention in the early time of entering the hypobaric hypoxia environment of the high altitude.

Objective To investigate clinical efficacy and prognostic factors of hilar cholangiocarcinoma.Methods The retrospective case-control study was conducted.The clinicopathological data of 322 patients with hilar cholangiocarcinoma who were admitted to the Xiangya Hospital of Central South University between December 2005 and November 2015 were collected.Preoperative staging and classification of tumor and treatment planning were carried out according to the results of laboratory and imaging examinations.Observation indexes:(1) clinical features and results of assisted examinations;(2) treatments and results of pathological examination;(3) followup and survival;(4) prognostic factors analysis:gender,age,preoperative highest total bilirubin (TBil),preoperative carcinoembryonic antigen (CEA),preoperative CA19-9,preoperative CA242,preoperative CA125,treatment methods and TNM staging.The follow-up of outpatient examination and telephone interview was perfornmed to detect patients' survival up to November 2016.Survival curve was drawn using the Kaplan-Meier method.Survival and univariate analyses were done using the Log-rank test,and multivariate analysis was done using the Cox proportional hazard model.Results (1) Clinical features and results of assisted examinations:among the 322 patients,there were 301 patients with a chief complaint of jaundice.Of the 322 patients,the preoperative highest levels of TBil,DBil,ALT and AST in 322 patients were 3.9-785.2 μmol/L,1.6-410.2 μ mol/L,14.8-484.5 U/L and 21.4-539.8 U/L,respectively.Levels of ALP and GGT in 272 patients were 93.8-1 890.0 U/L and 2.0-1 832.8 U/L,respectively.Seventy-seven of 292 patients had an elevated CEA level,272 of 298 patients had an elevated CA19-9 level,153 of 260 patients had an elevated CA242 level and 86 of 260 patients had an elevated CA125 level.According to Bismuth-Corlette type,24 patients were detected in type Ⅰ,115 in type Ⅱ,55 in type Ⅲa,63 in type Ⅲb and 65 in type Ⅳ.(2) Treatments and results of pathological examination:Of the 322 patients,104 patients underwent radical resection,including 79 with hilar bile duct resection (9 combined with vascular resection and reconstruction) and 25 with extended hepatic lobectomy (16 combined with caudate lobectomy),and 218 patients underwent palliative treatments,including 134 with external biliary drainage and 84 with internal biliary drainage.Five patients were dead in the perioperative period,of which 2 died of acute liver failure,1 died of systemic infection and multiple organ failure,1 died of acute renal failure and 1 died of acute suppurative cholangitis,septic shock and disseminated intravascular coagulation.Of 263 patients receiving pathological examination,adenocarcinoma was detected in 253 patients (12 with high-differentiated adenocarcinoma,85 with moderate-differentiated adenocarcinoma,33 with low-differentiated adenocarcinoma and 123 with indefinite differentiation),mucinous adenocarcinoma in 5 patients,cholangiocarcinoma in 3 patients and neuroendocrine carcinoma in 2 patients.TNM staging of 322 patients:stage Ⅰ was detected in 8 patients,stage Ⅱ in 53 patients,stage Ⅲ in 132 patients,stage Ⅳ in 96 patients and indefinite stage in 33 patients.(3) Follow up and survival:among the 322 patients,296 were followed up for 12-132 months,with a median follow-up time of 65 months,including 94 with radical resection and 202 with palliative treatments.Among the 296 patients,the median survival time and 1-,3-,5-year survival rates were 10 months,47.1％,20.2％ and 9.5％,respectively.0f296 patients with follow-up,median survival time and 1-,3-,5-year survival rates were 31 months,84.0％,46.2％,25.0％ in 94 patients receiving radical resection and 7 months,29.9％,8.1％ and 2.3％ in 202 patients receiving palliative treatment,respectively,with a statistically significant difference between the 2 groups (x2=78.777,P＜ 0.05).Among the 94 patients receiving follow-up and radical resection,the median survival time and 1-,3-,5-year survival rates were 31 months,82.1％,45.1％,25.7％ in 73 patients undergoing hilar bile duct resection and 35 months,90.5％,49.8％,22.1％ in 21 patients undergoing hepatic lobectomy,respectively,with no statistically significant difference (x2=0.186,P＞0.05).Among the 73 patients undergoing hilar bile duct resection,median survival time and 1-,3-,5-year survival rates were 16 months,57.1％,0,0 in 7 patients combined with vascular resection and reconstruction and 34 months,84.6％,49.5％,27.5％ in 66 patients undergoing simplex hilar bile duct resection,respectively,showing a statistically significant difference (x2 =11.977,P＜ 0.05).(4) Prognostic factors analysis:results of univariate analysis showed that preoperative highest TBil,preoperative CEA,preoperative CA242,preoperative CA125,treatment methods and TNM staging were related factors affecting prognosis of patients with hilar cholangiocarcinoma (x2=25.009,18.671,9.359,33.628,94.729,77.136,P＜0.05).Multivariate analysis showed that preoperative highest TBil ≥ 342.0 μmol/L,preoperative CEA ≥ 5.00 μg/L,palliative treatments,TNM stage Ⅲ and Ⅳ were the independent risk factors affecting the poor prognosis of patients with hilar cholangiocarcinoma (HR =2.270,2.147,3.166,2.351,95％ confidence interval:1.587-3.247,1.446-3.188,2.117-4.734,1.489-3.712,P＜0.05).Conclusions Prognosis of hilar cholangiocarcinoma is still unsatisfactory.The R0 resection is the key in radical surgery.Preoperative highest TBil≥342.0 μmol/L,preoperative CEA ≥ 5.00 μg/L,palliative treatments,TNM stage Ⅲ-Ⅳ are independent risk factors affecting the poor prognosis of patients with hilar cholangiocarcinoma.

Objective: To explore sarcoplasmic reticulum ryanodine receptor2 (RyR 2) expression and calcium releasing function in chronic heart failure (CHF) rabbits and to study the impact of long term valsartan treatment in relevant animals. Methods: HF model was established by volume overloading with pressure overloading in experimental rabbits. 27 rabbits were divided into 3 groups: Sham group, HF group and HF+valsartan group. n=9 in each group and the animals were treated for 7 weeks. Left ventricular structure, hemodynamic parameters, expression and functional changes of myocardiocyte sarcoplasmic reticulum RyR 2 were observed and compared among different groups. Results: Compared with Sham group, HF group had increased left ventricular mess index (LVMI), left ventricular end diastolic pressure (LVEDP) and decreased left ventricular shortening fraction, LVEF, all P<0.05. Compared with HF group, HF+valsartan group showed decreased LVMI, LVEDP and increased left ventricular shortening fraction, LVEF, all P<0.05. Sarcoplasmic reticulum RyR 2 expression and calcium releasing function were lower in HF group than Sham group, P<0.05; while they were both higher in HF+valsartan group than HF group, P<0.05. Conclusion: Long term application of valsartan could improve the cardiac function which might be related to increased myocardial sarcoplasmic reticulum RyR 2 expression and calcium releasing function in experimental CHF rabbits.

Objective:To explore the mechanism of metformin on osteogenic differentiation of rat jaw bone marrow mesenchymal cells(BMSCs)under high sugar and high fat environment.Methods:BMSCs of male SD rats were isolated and cultured in vitro and divided into 4 groups:Control group(C group),high sugar and high fat group(H group),control cells and H group cells were respectively stimulated with metformin(CM group and HM group).EDU staining was used to detect the cell proliferation;after osteogenesis and adi-pogenesis induction of BMSCs,alkaline phosphatase staining,alizarin red(O)staining and oil red O staining were respectively used to detect the osteogenic and adipogenic differentiation of the cells.The protein expression of Runx2 and OCN was detected by Western blot and immunofluorescence staining respectively.Results:Compared with the H group,in the HM group the cell proliferation ability was enhanced,the expression levels of Runx2 and OCN were increased(P＜0.05),the osteogenic differentiation ability was significantly in-creased,and the adipogenic differentiation ability was significantly decreased.Conclusion:In vitro,metformin promotes the prolifera-tion and osteogenic differentiation of rat jaw BMSCs cultured with a high sugar and high fat environment by increase of the expression of Runx2 and OCN.

To investigate the effect of hSCGF-alpha on human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs), we obtained hSCGF-alpha using genetic engineering, hSCGF-alpha gene was amplified from hUCMSCs cDNA using two-step PCR and was inserted into pET-28a(+) plasmid vector. Induced by IPTG at 20 degrees Celsius for 24 h, the fusion protein expressed in E. coli BL21 (DE3) was mainly existing in soluble form. The recombinant hSCGF-a was purified using NI-NTA affinity chromatography and the purity was up to 90%. The colony forming test revealed that combined use hSCGF-alpha and rmGM-CSF (recombinant murine GM-colony stimulating factor, rmGM-CSF) had granulocyte/macrophage (GM) promoting effects on murine bone marrow GM progenitor. In addition, the results indicated that hSCGF-alpha and rhGM-CSF had stimulatory effect on hUCMSCs and their synergetic effect was the strongest.
Cell Proliferation ; drug effects ; Cells, Cultured ; Cloning, Molecular ; Drug Synergism ; Escherichia coli ; genetics ; metabolism ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; Stem Cell Factor ; biosynthesis ; genetics ; Umbilical Cord ; cytology

Objective: To explore the changes of protein expression and activity of calcium/calmodulin-dependent protein kinase-II (CaMK-II) in myocardium nucleus and sarcoplasmic reticulum in experimental rabbits with heart failure (HF).
Methods: A total of 16 rabbits were divided into 2 groups: Sham group and HF group, the HF model was established by volume overload plus pressure overload.n=8 in each group and all animals were treated for 7 weeks. Left ventricular structure, hemodynamic parameters and protein expression and activity of CaMK-II in myocardium nucleus and sarcoplasmic reticulum were examined and compared between 2 groups.
Results: Compared with Sham group, HF group presented increased left ventricular mass index (LVMI) (1.32 ± 0.06) g/kg vs (3.61 ± 0.09) g/kg, LVEDP (-1.50 ± 0.50) mmHg vs (23.00 ± 2.37) mmHg, allP<0.05; while decreased left ventricular shorten fraction (37.83 ± 3.58) % vs (17.38 ± 3.13) % and LVEF (71.92 ± 4.56) % vs (38.50 ± 6.07) %, allP<0.05. The protein expression and activity of CaMK-II were both higher in HF group than Sham group, allP<0.05.
Conclusion: Increased protein expression and activity of CaMK-II in myocardium nucleus and sarcoplasmic reticulum might be one of the mechanisms for HF occurrence in experimental rabbits.

Objective To explore the effect of Yizheng Recipe on pyroptosis of alveolar epithelium in rats with recurrent respiratory tract infection(RRTI)based on HMGB1/TLR4/NF-κB signal pathway.Methods SD rats were firstly grouped into normal group and model group.In the model group,the RRTI rat model of spleen-Qi deficiency and lung- Qi deficiency was induced by fatigue combined with diet disorder and shavings and tobacco leaf smoking.Then,the rats in the model group were grouped into:RRTI model group(gavaged with normal saline 0.5 mL·100 g-1·d-1)and Yupingfeng Powder control treatment group(gavaged with Yupingfeng Powder suspension 0.5 mL·100 g-1·d-1),Yizheng Recipe high,medium and low-dose treatment groups(gavaged with Yizheng Recipe 0.75,0.5,0.25 mL·100 g-1·d-1),the normal group(gavaged with normal saline 0.5 mL·100 g-1·d-1).The general activities of the rats were observed and the pulmonary function of the rats was detected by the AniRes2005 animal pulmonary function analysis system;the expression of inflammatory factors in rat serum was detected by ELISA;HE,PAS and TUNEL staining were applied to detect the pathological changes of lung tissue;Western blot was applied to detect cell pyroptosis and the expression of HMGB1/TLR4/NF-κB signaling pathway-related proteins.Results Compared with the normal group,the lung function of the RRTI model group was weakened;the inflammatory response increased,and the lung tissue damage,edema and apoptosis were obviously increased;meantime,the pyroptosis was aggravated,and the HMGB1/TLR4/NF-κB pathway was activated(P<0.05).Compared with the RRTI model group,the Yupingfeng Powder control treatment group and the Yizheng Recipe high-dose,medium-dose and low-dose treatment groups all had different degrees of remission in lung function;inflammatory response reduced,the lung tissue damage,edema and apoptosis reduced;meantime,the pyroptosis and HMGB1/TLR4/NF-κB pathway were inhibited(P<0.05).Yizheng Recipe high-dose treatment group and Yupingfeng Powder control treatment group had similar curative effect on RRTI model rats(P>0.05);and Yizheng Recipe was dose-dependent in treating RRTI model rats(P<0.05).Conclusion Yizheng Recipe may inhibit the pyroptosis of alveolar epithelial cells in RRTI model rats by reducing the inflammatory response,and then protecting the lung tissue from damage,this process may be related to the HMGB1/TLR4/NF-κB pathway.

OBJECTIVE： To establish UPLC-Q- TOF-MS/MS fingerprint of Fritillaria thunbergii ， and to define its anti-inflammatory quality markers. METHODS ：The determination was performed on Eclipse Plus C 18 column with mobile phase consisted of methanol- 0.1％ formic acid solution （gradient elution ）at the flow rate of 0.4 mL/min. The column temperature was set at 30 ℃，and injection volume was 2 µL. The electrospray ion source was used to scan in the range of m/z 50-1 200 with positive and negative ion detection mode. UPLC-Q-TOF-MS/MS fingerprints of 10 batches of F. thunbergii from different habitats were established by using the Similarity Evaluation System of TCM Chromatographic Fingerprints （2004A edition ）. With ear swelling degree，the serum levels of MDA and NO as anti-inflammatory indexes ，the correlation between the relative area of common peaks in fingerprint and the anti-inflammatory indexes was analyzed by using bivariate correlation analysis and grey correlation analysis ， and the quality markers were screened and identified. RESULTS ：In positive and negative ion mode ，10 batches of F. thunbergii had 26 peaks and 10 peaks. Based on bivariate correlation analysis and grey correlation analysis ，nine quality markers related to anti-inflammatory effect were found ，which were identified as peiminine ，peimine，cyclobalamine，daucidin，polyphyllin Ⅴ， bitumen podophyllotoxin ，phytosterol，ent-kaur-15-en-17-ol，ent-17-norkauran-16-one. CONCLUSIONS ：Established UPLC-Q- TOF-MS/MS fingerprint can be used to evaluate the quality of F. thunbergii ；peiminine，peimine and cyclobalamine and so on may be the quality markers of anti-inflammatory effect of F. thunbergii .

Objective To explore the changes in serum energy metabolites in patients with peripheral T-cell lymphoma,and investigate serum biomarkers for monitoring peripheral T-cell lymphoma from the perspective of energy metabolism.Methods Multiple/selected reaction monitoring(MRM/SRM)was used to detect the energy-related metabolites in the sera of 16 patients with newly diagnosed peripheral T-cell lymphoma admitted in the Hematology Medical Center of the Second Affiliated Hospital of Army Medical University from November 2020 to December 2021,as well as 10 recruited healthy volunteers.The corresponding clinical data including medical history,laboratory results and image data were collected and retrospectively analyzed.Results Significant differences were seen in the contents and expression profiles of serum energy metabolism-related products between the patients and the healthy volunteers.The patients had significantly reduced serum contents of cyclic AMP,succinate,citrate and cis-aconitate(P＜0.05),and elevated D-glucose 6-phosphate content(P＜0.05).The serum contents of citrate and succinate were negatively correlated with the risk stratification(low-,moderate-and high-risk)and clinical stage of the disease(P＜0.05).Meanwhile,there was a negative correlation between the contents of L-malic acid and citrate and the mid-term efficacy evaluation results,such as complete/partial response(CR/PR)or stable disease(SD)(P＜0.05).For patients with extranodal NK/T cell lymphoma(n=10),there were also significant reductions in the contents of cyclic AMP,succinate,citrate,isocitrate and cis-aconitate in the sera of patients compared with healthy volunteers(P＜0.05),and the contents of citrate and succinate were negatively correlated with the clinical stage(P＜0.05)and were rather correlated with mid-term efficacy evaluation results(CR/PR or SD)(P＜0.05).For patients with angioimmunoblastic T-cell lymphoma(n=6),the serum contents of cyclic AMP,citrate and succinate were significantly lower,while the content of D-glucose 6-phosphate was higher when compared with the healthy volunteers(P＜0.05),and the content of succinate was negatively correlated with both clinical stage and risk grade of the patients(P＜0.05).Conclusion There are 5 serum differential metabolites identified between patients with peripheral T-cell lymphoma and healthy controls,and succinate and citrate are expected to be serum biomarkers of peripheral T-cell lymphoma.