Objective To investigate the effects of mesenchymal stem cells(MSCs) on the recovery of gentamicin-induced acute renal failure(ARF),and explore its possible mechanism.Methods MSCs were isolated from a male SD rat and labeled with DAPI before injection.ARF rat model was reproduced by hypodermic injection of gentamicin.At the time that gentamicin was injected,MSCs were injected through tail vein.Thirty SD rats were randomly divided into three groups(10 each): MSCs injection group(group M),DMEM-F12 infusion group(group D) and control group(group C).Rats were sacrificed on day 7 after gentamicin injection.The levels of serum Scr and BUN were determined,and the renal tissue was sampled for pathological observation.Cell apoptotic index(AI) was calculated after terminal deoxynucleotidyl transferase mediated d-UTP nick end labeling(TUNEL).The expressions of Bcl-2,Fas and FasL proteins and PCNA were determined by immunohistochemical technique.The distribution of DAPI positive MSCs in kidney of recipient SD rats was examined by immunohistochemistry.Results HSK score(5.5?0.8 vs 10.2?0.8,P

Objective To investigate the effects of preconditioning with repeated electroacupuncture (EA)at Shenshu acupoint on renal ischemia-reperfusion (I/R) injury in rats.Methods Fifty adult male Sprague-Dawley rats,weighing 260-320 g,were randomly divided into 4 groups:sham operation group (n =5),I/R group,pentobarbital sodium + I/R group (PB + I/R group) and repeated EA at Shenshu acupoint + I/R group (EA + I/R group).The model of renal I/R injury was established by occlusion of bilateral renal pedicles for 45 min followed by reperfusion.Pentobarbital sodium 30 mg/kg was injected intraperitoneally everyday for 5 consecutive days and I/R was induced 24 h after the last injection in PB + I/R group.The animals received EA at Shenshu acupiont 30min per day for 5 consecutive days under pentobarbital sodium anesthesia and I/R was induced 24 h after the last preconditioning in EA + I/R group.Blood samples were taken at 1,3 and 7 days after I/R to determine the levels of serum blood urea nitrogen (BUN) and creatinine (Cr).The animals were then sacrificed and the kidney was isolated.The histological changes of the kidney was scored.The apoptosis in renal tubular epithelial cells was measured using TUNEL at 3 days after I/R.Apoptosis index (AI) was calculated.The expression of proliferating cell nuclear antigen (PCNA),Bcl-2,Bax,Fas and FasL in renal tubular epithelial cells was measured by immuno-histochemistry at 3 days after I/R.Results Compared with group S,the levels of serum BUN and Cr,and histological score were significantly increased at 1,3 and 7 days after I/R in I/R and PB + I/R groups,and at 1 and 3 days after I/R in EA + I/R group,the expression of PCNA,Bcl-2,Bax,Fas and FasL was up-regulated in I/R,PB + I/R and EA + I/R groups,and Bax/Bcl-2 ratio was significantly increased in I/R and PB + I/R groups (P ＜0.05).Compared with I/R and PB + I/R groups,the levels of serum BUN and Cr,and histological score were significantly decreased at 3 and 7 days after I/R,AI and the expression of Bax,Fas and FasL were significantly decreased,and the expression of PCNA and Bcl-2 was up-regulated in EA + I/R group (P ＜ 0.05).Conclusion Preconditioning with repeated EA at Shenshu acupoint can attenuate the renal I/R injury in rats by promoting the proliferation of renal tubular epithelial cells and reducing the apoptosis in cells.

Human cytomegalovirus (HCMV) late mRNA expression in megakaryoblast and in turn the pathogenesis of idiopathic thrombocytopenic purpura (ITP) patients with HCMV infection, and effectiveness of ganciclovir were investigated. Colony forming unit-megakaryocytes (CFU-MK) of 46 ITP patients with HCMV infection were incubated from patients' bone marrow mononuclear cells (MNC). Reverse transcriptase-polymerase chain reaction (RT-PCR) was subsequently used for CFU-MK for HCMV-late mRNA detection. Ganciclovir therapy was given to both HCMV-late mRNA positive and negative groups for comparison of therapeutic effectiveness. The results in 19 of 46 CFU-MK culture cells specimens with positive HCMV-DNA by PCR or positive CMV-IgM by enzyme linked immunosorbent assay (ELISA) in the correspondent serum of peripheral blood were positive for HCMV-late mRNA. Sixteen out of 19, patients with positive HCMV-late mRNA CFU-MK had a positive response to ganciclovir. Amongst 27 patients with negative HCMV-late mRNA CFU-MK, only 4 positive responders to ganciclovir therapy were observed. Curative effectiveness of ganciclovir in HCMV-late mRNA positive group was significantly higher than that in HCMV-late mRNA negative group (P<0.01). It was suggested that HCMV could directly infect CFU-MK, which might be one of the mechanisms responsible for HCMV related ITP. The ganciclovir is an effective therapy in resulting in the increases in thrombocyte in the ITP patients whose HCMV- late mRNA was positive in their CFU-MK.

BACKGROUND:Abnormal immunological function is possibly observed after cardiac valve replacement. However, effect of continuous venovenous hemofiltration on immunological function after cardiac valve replacement is rarely reported.
OBJECTIVE:To observe the effect of continuous venovenous hemofiltration on the cellular immune function in patients with multiple organ dysfunction syndrome after cardiac valve replacement.
METHODS:Thirty-one patients with multiple organ dysfunction syndrome after cardiac valve replacement in Xijing Hospital, the Fourth Military Medical University of Chinese PLA, from August 2008 to July 2009, were included in this study. They were treated with continuous venovenous hemofiltration using AV600 hemofilter and were divided into two groups:survival group (17 survivors) and death group (14 deaths). In addition, 16 healthy blood donors served as the control group.
RESULTS AND CONCLUSION:The duration of acute renal failure before continuous venovenous hemofiltration of survival group was significantly lower than that of death group (P<0.05). Before continuous venovenous hemofiltration, the CD4+/CD8+ratio and Th1/Th2 ratio of survival group and death group were lower than that of control group (P<0.05), and lymphocyte apoptosis rate and Fas/FasL antigen expression were higher than that of control group (P<0.05). This evidence suggested the presence of immunosuppression state and Th1/Th2 disbalance. During continuous venovenous hemofiltration, the CD4+/CD8+ratio and Th1/Th2 ratio of survival group were gradual y increased (P<0.05), while lymphocyte apoptosis rate and Fas/FasL antigen expression were gradual y decreased (P<0.05) at 24 hours. The above changes were observed in both survival group and death group, but emerged latter in death group. Continuous venovenous hemofiltration can improve cellular immune function, maintain the balance of T lymphocyte subsets and Th1/Th2, down-regulate Fas/FasL expression on the surface of lymphocyte membrane, and decrease lymphocyte apoptosis rate in patients with multiple organ dysfunction syndrome after cardiac valve replacement. Early continuous venovenous hemofiltration can improve patients’ prognosis.

Hunan University of Chinese Medicine has improved the implementation of animal ethics education from the aspects of carrying out elective courses, emphasizing the implementation of pre-class learning, integrating ethics education into experimental teaching, cultivating students' good medical literacy in a subtle way, and further deepening their learning by offering elective courses of animal ethics. The results show that the opening of elective courses and the enforcement of experimental animal ethics education in experimental classes have greatly strengthened the students' experimental animal ethics, which is conducive to the formation of students' medical literacy.

Human cytomegalovirus (HCMV) late mRNA expression in megakaryoblast and in turn the pathogenesis of idiopathic thrombocytopenic purpura (ITP) patients with HCMV infection, and effectiveness of ganciclovir were investigated. Colony forming unit-megakaryocytes (CFU-MK) of 46 ITP patients with HCMV infection were incubated from patients' bone marrow mononuclear cells (MNC). Reverse transcriptase-polymerase chain reaction (RT-PCR) was subsequently used for CFU-MK for HCMV-late mRNA detection. Ganciclovir therapy was given to both HCMV-late mRNA positive and negative groups for comparison of therapeutic effectiveness. The results in 19 of 46 CFU-MK culture cells specimens with positive HCMV-DNA by PCR or positive CMV-IgM by enzyme linked immunosorbent assay (ELISA) in the correspondent serum of peripheral blood were positive for HCMV-late mRNA. Sixteen out of 19 patients with positive HCMV-late mRNACFU-MK had a positive response to ganciclovir. Amongst 27 patients with negative HCMV-late mRNA CFU-MK, only 4 positive responders to ganciclovir therapy were observed. Curative effectiveness of ganciclovir in HCMV-late mRNA positive group was significantly higher than that in HCMV-late mRNA negative group (P＜0.01). It was suggested that HCMV could directly infect CFU-MK, which might be one of the mechanisms responsible for HCMV related ITP. The ganciclovir is an effective therapy in resulting in the increases in thrombocyte in the ITP patients whose HCMV- late mRNA was positive in their CFU-MK.

TCM experimental teaching has a very important position in university training process, in which the laboratory standardized management and the strengthening of the teaching quality monitoring play important roles. According to the the experimental teaching reforms and requirements enacted in recent years by the Ministry of Education of the People’s Republic of China, Hunan TCM University has conducted a series of standardized management of exploration and practice in the laboratory.

Objective:To study the effect of Compound Duzhong Jiangu Granules on joint function, quality of life and inflammatory factors in patients with Kashin-Beck disease.Methods:By group design, 135 patients with Kashin-Beck disease in Chongxin County, Pingliang City, Gansu Province, were selected and divided into intervention group (100 cases treated with Compound Duzhong Jiangu Granules, 12 g/bag, 1 bag/time, 3 times/day, treatment for 1 month) and control group (35 cases treated with ibuprofen, 0.3 g/capsule, 1 capsule/time, 2 times/day, treatment for 2 weeks) according to the randomized, single-blind principle. The changes of joint dysfunction index score, joint function improvement rate, total scores of Kashin-Beck disease quality of life (KBDQOL) and scores of each dimension were analyzed before treatment, 1 month and 3 months after treatment. The serum levels of interleukin-6 (IL-6), nuclear factor κB (NFκB)-p65, inducible nitric oxide synthase (NOS2), nitric oxide (NO), and prostaglandin E2 (PGE2) were detected by enzyme-linked immunosorbent assay before treatment and 1 month after treatment.Results:The total scores of joint dysfunction index of the intervention group and the control group 1 month after treatment and 3 months after treatment were lower than those before treatment, but 3 months after treatment was higher than 1 month after treatment ( P < 0.001). One month after treatment, the total effective rates of joint function improvement in the intervention group and the control group were 68.00% (68/100) and 54.55% (18/33), respectively; 3 months after treatment, the total effective rates of the intervention group and the control group were 36.00% (36/100) and 39.39%(13/33), respectively. The total scores and scores of each dimension of KBDQOL were not significant for the main effect of the group ( P > 0.05), but significant for the main effect at the time point ( P < 0.05), and there was no interactive effect ( P > 0.05). There were significant differences in the scores of social support and mental status dimensions in the intervention group at different time points ( P < 0.001). And in the intervention group, the total score, the scores of physical function, activity limitation, economy and overall health dimensions were statistically significant between before treatment and 1 month after treatment, 1 month after treatment and 3 months after treatment( P < 0.05); however, there was no significant difference between before treatment and 3 months after treatment ( P > 0.05). One month after treatment, the serum PGE2 levels of both groups were decreased ( P < 0.05), and there was no difference in other inflammatory factors at different groups and time points ( P > 0.05). Conclusion:Compound Duzhong Jiangu Granules can effectively inhibit the inflammatory reaction of patients with Kashin-Beck disease, promote the improvement of joint function and improve the quality of life in various aspects.

OBJECTIVE：To study the intervention effects of Huan gqi injecti on on leucopenia model mice. METHODS ： Kunming mice were randomly divided into normal group ，model group and drug group ，with 8 mice in each group. Model group and drug group were given intraperitoneal injection of cyclophosphamide to induce leukopenia model. Normal group was given intraperitoneal injection of equal volume of sterile water. After modeling successfully ，normal group and model group were given intraperitoneal injection of equal volume of sterile water ；drug group was given intraperitoneal injection of Huangqi injection 0.04 mL/10 g，once a day ，for consecutive 7 d. Based on the detection of blood routine indexes （leukocyte，lymphocyte，neutrophil， monocyte count ）of mice in each group ，the metabolites in serum were analyzed by LC-MS. Principal component analysis （PCA）， partial least squares discriminant analysis （PLS-DA），orthogonal partial least squares-discriminant analysis （OPLS-DA），HMDB， METLIN， KEGG and other databases as well as related literatures were used to identify the differential metabolites. The metabolic pathways of differential metabolites were analyzed with MetPA online tools ，and the correlation of blood routine indexes and differential metabolites were analyzed on the basis of Pearson correlation coefficient. RESULTS： Compared with normal group ， blood rountine indexes of model group were decreased significantly （P＜ 0.01）. Compared with model group ，above blood r ountine indexes of drug group were all increased siginificantly （P＜0.01）. LC-MS chromatogram of serum samples in normal group and model group were significantly different ，and LC-MS metabonomics data of serum samples in drug group were similar to those of normal group. Multivariate statistical analysis and correlated database analysis revealed that compared with normal group ，serum contents of 17 metabolites as L-isoleucine，eicosapentaenoic acid were increased significantly in model group ，while the contents of 4 metabolites as spermidine were decreased significantly （P＜0.05 or P＜0.01）. Compared with model group ，Huangqi injection could reverse the serum contents of 9 metabolites in mice ，such as citric acid ，L-proline，acetylcarnitine，L-isoleucine， L-phenylalanine，sphingosine-1-phosphate，lysophosphatidylinositol，eicosapentaenoic acid and linoleic acid （P＜0.05 or P＜ 0.01），which were associated with linoleic acid metabolism ，biosynthesis of phenylalanine ，tyrosine and tryptophan ，and phenylalanine metabolism （metabolism pathway influential values were all higher than 0.1）. Correlation analysis showed that there was a significant correlation between blood routine indexes and the contents of D-sphingosine，linoleic acid and citric acid in model group（the absolute values of r were generally greater than 0.5）. CONCLUSIONS ：Huangqi injection can increase the counts of leukocytes，lymphocytes，neutrophils and monocytes in leucopenia model mice. The increase of leukocytes may be related to linoleic acid metabolism ，biosynthesis of phenylalanine ，tyrosine and tryptophan ，and phenylalanine metabolism.