Objective To study the endogenous metabolites of liver-kidney deficiency syndrome in knee-joint osteoarthritis (KOA) and explore the metabolic profile of KOA liver-kidney deficiency syndrome. Methods Totally 50 cases of KOA with liver-kidney deficiency syndrome and 50 cases of KOA with non-liver-kidney deficiency syndrome were collected respectively, and 20 cases of healthy volunteers were collected as the normal group. The serum samples of subjects were collected after fasting for 8 h. Hydro-Nuclear Magnetic Resonance (1H-NMR) spectrometer was collected. Principal component analysis and partial least squares discrimination analysis were used to conduct multivariate statistical analysis. Results 1H-NMR could identify 23 kinds of metabolites, and there was statistical significance between KOA patients and healthy volunteers (P<0.05, P<0.01). There was statistical significance in cartilage matrix metabolism, energy metabolism, lipid metabolism, pain, inflammation-related metabolites in patients with KOA liver-kidney deficiency syndrome and patients with KOA non-liver-kidney deficiency syndrome (P<0.05). Conclusion Patients with KOA liver-kidney deficiency syndrome have a unique 1H-NMR metabolic profile, KOA syndrome has a metabolic material basis.

BACKGROUND:Mesenchymal stem cells are found to have the immunoregulatory activities and a potential application prospect in the treatment of autoimmune diseases.
OBJECTIVE:To explore the mechanism of transplanting mesenchymal stems cells on the treatment of multiple sclerosis.
METHODS:The mouse mesenchymal stems cells were prepared, and injected into the al ogenic and syngenic normal mice, to detect the frequency of CD4+CD25+Foxp3+T cells in the spleen, thymus, and lymph nodes by flow cytometry, and to detect the Foxp3, transforming growth factor-β1, and interleukin-10 mRNA in the spleen, thymus, and lymph nodes by reverse transcription-PCR.
RESULTS AND CONCLUSION:Transplantation of mesenchymal stem cells on normal mice led to a significant up-regulation of CD4+CD25+Foxp3+T cells, Foxp3, transforming growth factor-β1, and interleukin-10 mRNA in the spleen, thymus, and lymph nodes both in the al ogenic and syngenic transplant groups. Transplantation of mesenchymal stem cells may be an available method in the treatment of autoimmune diseases, and CD4+CD25+Foxp3+T cell, Foxp3, transforming growth factor-β1, and interleukin-10 may be involved in this process.

This study examined the potential antilithic effects of a traditional Chinese medicine Urtica dentata Hand (UDH) in experimental rats and screened the optimal extract of UDH as a possible therapeutic agent for kidney stones. The rat model of urinary calcium oxalate stones was induced by intragastric (i.g.) administration of 2 mL of 1.25% ethylene glycol (EG) and 1% ammonium chloride (AC) for 28 days and was confirmed by Color Doppler ultrasound imaging. The rats in different experimental groups were then intragastrically given petroleum ether extract (PEE), N-butanol extract (NBE), aqueous extract (AqE) of UDH, Jieshitong (positive control drug), and saline, respectively. Treatment with NBE significantly reduced the elevated levels of urinary calcium, uric acid, phosphate, as well as increased urinary output. Accordingly, the increased calcium, oxalate levels and the number of calcium oxalate crystals deposits were remarkably reverted in the renal tissue of NBE-treated rats. In addition, NBE also prevented the impairment of renal function to decrease the contents of blood urea nitrogen (BUN) and creatinine. Taken together, these data suggest that NBE of UDH has a beneficial effect on calcium oxalate urinary stones in rats by flushing the stones out and protecting renal function.

Objective This study was performed to investigate the effect of hypoxia on glucose-6-phosphate isomerase (G6PI) expression and cell cycle of fibroblast-like synoviocytes from synovium of rheumatoid arthritis (RA) and osteoarthritis (OA) under hypoxia or normoxia.Methods Fibroblast-like synoviocytes were cultured with either of hypoxia (3％ oxygen) or normoxia (21％ oxygen) for 24 hours.The mRNA expression of G6PI and HIF-1α was tested by PCR quantification,while the protein levels of G6PI and HIF-1α were measured by western blot.Cell cycle was performed by FACS.T-test and Mann-Whitney U were used for statistical analysis.Results The expression levels of G6PI mRNA under hypoxia in RA were higher than those of OA (2.6±0.4 vs 1.5±0.4,P＜0.05).The protein levels of G6PI in RA were higher than those of OA (P＜0.05).The expression levels of HIF-1α mRNA under hypoxia in RA were higher than those of OA (2.9±0.8vs 1.4 ±0.4,P＜0.05).The protein levels of HIF-1α in RA were higher than those of OA (P＜0.05).The G1 phase ratio of cell cycle was decreased significantly under hypoxia than those of normoxia in RA ELs (t=1 1.31,P＜0.05).The S and G2 phase ratio of cell cycle were increased.Conclusion Hypoxia upregulates G6PI and HIF-1α expression and improves proliferation in fibroblast-like synoviocytes.

Objective To investigate the influence of classically activated macrophage (M1) on the proliferation of rheumatoid arthritis (RA) fibroblast-like synovial (FLS) and osteoarthritis (OA) FLS proliferation.Methods Human monocytes leukemia cells (THP)-1 were induced into M1 by lipopolysaccharides (LPS) and interferon gamma (IFN-γ),M1 specific surface molecular markers human leukocyte antigen (HLA)-DR and CD197 were detected by flow cytometry (FCM).RA-FLS and OA-FLS were co-cultured with M1 by transwell chambers,the proliferation of RA-FLS and OA-FLS were observed by crystal violet staining assay.MTS was used to detect cytokines secreted from M1 on the multiplication of RA-FLS and OA-FLS.TNF-α and IL-12 were detected by enzyme linked immunosorbent assay (ELISA).Paried student t test was used for statistical analysis.Results THP-1 were induced into M1 by LPS and IFN-γ,the expression rates of M1 surface specific molecular markers HLA-DR and CD197 were 78.25％ and 87.96％.Crystal violet staining showed that RA-FLS and OA-FLS proliferation were significantly inhibited after co-cultured with M1 48 h,RA-FLS and OA-FLS of each vision under microscope in co-culture groups were (64 ±30) and (85 ±23) respectively,while the RA-FLS and OA-FLS in separate culture groups were (467±87) and (263±78) respectively,the difference was statistically significant (t=7.459,3.791;P＜0.05).MTS assay indirectly reflected that the cytokines from M1 suppressed RA-FLS and OA-FLS proliferation (t=-7.155,-8.111;P＜0.05).The concentration of TNF-α in cell culture supernatants secreted from RA-FLS group and RA-FLS/M1 co-culture group respectively were (0.024±0.01 1) ng/ml and (0.832±0.241) ng/ml respectively,the concentration of IL-12 from the two groups were (0.033±0.015) ng/ml and (0.372±0.122) ng/ml respectively.TNF-α from OA-FLS and OA-FLS/M1 co-culture group respectively were (0.031±0.017) ng/ml and (0.852±0.323) ng/ml,IL-12 were (0.012±0.009) ng/ml,(0.373±0.144) ng/ml.Compared with FLS separate culture group,the concentration of TNF-α and IL-12 were obviously elevated (t=-4.997,-4.777,-4.407,-4.334;P were all ＜0.05).Conclusion M1 can significantly inhibite RA-FLS and OA-FLS proliferation,this may be related to the increased concentration of TNF-α and IL-1 β in from cell culture supernatant.

Objective To observe expression levels of N-methyl-D-aspartate (NMDA) receptor and cholecystokinin (CCK) in the hippocampus and spinal cord in morphine withdrawal or tolerance mice treated by electroacupuncture or catgut embedding and explore the difference between the regulating effects of electroacupuncture and catgut embedding on morphine withdrawal and tolerance.Methods Fifty-six male C57BL/6J mice were randomly allocated to withdrawal control, withdrawal model, withdrawal catgut embedding and withdrawal electroacupuncture groups, and tolerance control, tolerance model, tolerance catgut embedding and tolerance electroacupuncture groups, 7 mice in each group. A model of morphine withdrawal was made by subcutaneous injection of morphine hydrochloride using 7-day increasing addiction method. The withdrawal control group was injected with an equal volume of normal saline at the same time points. In the withdrawal electroacupuncture group, electroacupuncture at bilateral points Shenshu was performed using a Han’s acupoint nerve stimulation device (HANS-200) at 15 min after an injection of morphine hydrochloride. In the withdrawal catgut embedding group, 0.5 cm chromic catgut was embedded in bilateral points Shenshu at 15 min after an injection of morphine hydrochloride. Addiction was promoted by intraperitoneal injection of naloxone 4 mg/kg at 10 o’clock on the seventh day’s morning and Withdrawal reactions were observed in the mice. The score was recorded using the Ryuta Tomoji opioid withdrawal symptoms evaluation scale. NMDA receptor and CCK contents in the hippocampus and spinal cord were measured by enzyme-linked immunosorbent assay (ELISA). A model of morphine tolerance was made by subcutaneous injection of morphine 10 mg/kg. The tolerance control group was injected with tolerance normal saline 10 ml/kg at the same time. In the tolerance catgut embedding group, catgut was embedded in point Shenshu at the first day after model making. In the tolerance electroacupuncture group, point Shenshu was given electroacupuncture at the first day after model making. After seven days of treatment, NMDA receptor and CCK contents in the hippocampus and spinal cord were measured by ELISA.Results There were statistically significant differences in hippocampal NR2B and CCK expressions between the withdrawal model and withdrawal control groups (P<0.05). There was a statistically significant difference in hippocampal NR2B expression between the withdrawal electroacupuncture and withdrawal model groups (P<0.05). There was a statistically significant difference in hippocampal CCK expression between the withdrawal catgut embedding or withdrawal electroacupuncture group and the withdrawal model group (P<0.05). There were statistically significant differences in spinal cord NR2A, NR2B and CCK expressions between the withdrawal model and withdrawal control groups (P<0.05). There were statistically significant differences in spinal cord NR2A and NR2B expressions between the withdrawal electroacupuncture and withdrawal model groups (P<0.05). There were statistically significant differences in hippocampal NR2A, NR2B and CCK expressions between the tolerance model and tolerance control groups (P<0.05). There was a statistically significant difference in hippocampal CCK expression between the tolerance catgut embedding and tolerance model groups (P<0.05). There was a statistically significant difference in hippocampal NR1 expression between the tolerance electroacupuncture group and the tolerance model or tolerance catgut embedding group (P<0.05). There was a statistically significant difference in spinal cord CCK expression between the tolerance catgut embedding or withdrawal electroacupuncture group and the tolerance model group (P<0.05).Conclusions Both catgut embedding and electroacupuncture at point Shenshu have a reducing effect on morphine tolerance and withdrawal. The therapeutic effect of electroacupuncture is better than that of catgut embedding.

Objective Detecting antibodies against RBCs,platelets,lymphocytes,and neutrophils by a solid-phase antibodies screening system.Methods mono-layer blood cell immobilized in the bottom of U-microplate respectively to prepare the solid-phase antibody screening system.Then detecting the antibodies exist or not in 2 150 random blood donor and 440transfusion patients' samples.Analyze the influence of antibodies against blood cells to the transfusion effect.Results 53RBC antibodies,22 platelet antibodies,13 lymphocyte antibodies,55 neutrophil antibodies were detected in random blood donor samples;31 RBC antibodies,38 platelet antibodies,24 lymphocyte antibodies,43 neutrophil antibodies and 115 mixed (two or more antibodies against RBC,platelets,lymphocytes or neutrophils exist at the same time) were detected in transfusion patients' samples after detection.233 suspected adverse reaction happened after transfusion,which antibodies were detected in 133 samples.Conclusion The antibodies against blood cells solid-phase screening system can applied in pre-and post-transfusion detection.The solid-phase screening method is more sensitivity than serologic tube test and flow cytometry.Existing of antibodies influence the transfusion effect.

Objective:To study the amino acid and codon usage profile of HIV-1 Env gene in donors whose serum exhibit highly broad cross-neutralizing activity. Methods:The samples were divided into highly broad cross-neutralizing activity group (hBCN + group) and non-highly broad cross-neutralizing activity group (hBCN - group) based on whether the neutralization breadth was higher than 90% or not. Full-length Env genes were amplified by single genome amplification (SGA) method from patients′ plasma samples, and the characteristics of Env sequences in hBCN + group were compared with hBCN - group. The correspondence analysis (COA) on relative amino acid usage (RAAU), adaptability to host based on similarity index D( A, B) and relative synonymous codon usage (RSCU) values of Env genes (hBCN + and hBCN -) with respect to human host RSCU were analyzed. Results:Correspondence analysis showed that the RAAU data of hBCN + group and hBCN - group were distributed along the two main axes to form two relatively separated clusters, indicating that the Env genes of the two groups had relatively unique amino acid usage patterns; the similarity index calculation results showed that hBCN + group (0.097) was lower than the hBCN - group (0.102), in addition, the Env gene of the hBCN + group had less frequency of similarly selected codons with human host system compared to hBCN - group. Conclusions:Env genes in hBCN + group and hBCN - group may have relatively unique amino acid usage patterns, and virus strains in hBCN + group are less adaptable to the host than those in hBCN - group.

Objective　To explore the lethal effect of Bacillus thuringiensis israelensis (Bti) on the 3rd instar larvae of Aedes albopictus and the influence of environmental temperature. Methods Probit analysis bioassay was used to detect comparatively analyze the lethal effect of Bti on third instar larvae of Aedes albopictus under different environmental temperatures. The accumulated mortality rate of larvae after treatment with different Bti concentrations and corresponding P-values and concentration logarithm were used for linear regression analyses to obtain the linear regression formula, and important dose parameters such as LC50 and LC95 were calculated. Result Bti had a significant lethal effect on Aedes albopictus larvae and the drug efficacy was rapid. At a standard 28 ℃, significant larval mortality was observed 12 h after Bti treatment, and the cumulative mortality rate increased with the rise of treatment time and concentration of Bti. Linear regression formulae of the lethal effect of Bti on Aedes albopictus larvae at different time points and temperatures were obtained. The insecticidal effect was ideal between 28-32 ℃ for the same Bti concentration, and the insecticidal effect was relatively weakest in a low temperature environment of 20 ℃; the LC95 at 28 ℃ at each time point was the lowest, with LC95 values of 122.418 IU/L, 109.404 IU/L, 96.474 IU/L, and 92.229 IU/L at 24 h, 48 h, 72 h, and 96 h respectively. Conclusions Bti has a strong lethal effect on the third instar larvae of Aedes albopictus, environmental temperature can influence the insecticidal effect of Bti on Aedes albopictus, raising temperatures within a certain range can enhance the lethal effect; when using Bti in reality, the influence of temperature factors should be fully considered to achieve better insecticidal effect.

Analysis of neural stem cells' movements is one of the important parts in the fields of cellular and biological research. The main difficulty existing in cells' movement study is whether the cells tracking system can simultaneously track and analyze thousands of neural stem cells (NSCs) automatically. We present a novel cells' tracking algorithm which is based on segmentation and data association in this paper, aiming to improve the tracking accuracy further in high density NSCs' image. Firstly, we adopted different methods of segmentation base on the characteristics of the two cell image sequences in our experiment. Then we formed a data association and constituted a coefficient matrix by all cells between two adjacent frames according to topological constraints. Finally we applied The Hungarian algorithm to implement inter-cells matching optimally. Cells' tracking can be achieved according to this model from the second frame to the last one in a sequence. Experimental results showed that this approaching method has higher accuracy compared with that using the topological constraints tracking alone. The final tracking accuracies of average of sequence I and sequence II have been improved 10.17% and 4%, respectively.
Algorithms ; Animals ; Cell Count ; Cell Movement ; Cell Tracking ; statistics & numerical data ; Image Processing, Computer-Assisted ; methods ; Microscopy, Fluorescence ; Models, Theoretical ; Neural Stem Cells ; cytology