Quality evaluation of Crataegus pinnatifida leaves by fingerprint combined with quantitative analysis of multi-components by single-marker / 中国药房

OBJECTIVE To establish the fingerprint and multi-component content determination method of Crataegus pinnatifida leaves from different producing areas， and to evaluate the quality of C. pinnatifida leaves and screen the differential markers. METHODS Seventy-eight batches of C. pinnatifida leaves were collected from Chengde of Hebei Province， Huludao of Liaoning Province， Yuncheng of Shanxi Province and Linyi of Shandong Province. High-performance liquid chromatography （HPLC） and Similarity Evaluation System for Traditional Chinese Medicine Chromatographic Fingerprints （2012 edition） were used to draw the fingerprints and conduct similarity evaluation. Grey correlation analysis， cluster analysis （CA）， principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA） were performed by using SPSS 19.0， MetaboAnalyst 5.0 and SIMCA 14.1 software. The differential markers affecting the quality of C. pinnatifida leaves were screened with variable importance in the projection （VIP） value greater than 1 and the error line not exceeding the origin as the criterion. Using vitexin rhamnoside as an internal reference， the contents of chlorogenic acid， glucosylvitexin， hypericin and isoquercetin in 78 batches of C. pinnatifida leaves were determined by the same HPLC combined with quantitative analysis of multi- components by single-marker （QAMS）， and the results were compared with external standard method. RESULTS Eight common peaks were calibrated in the fingerprints for 78 batches of C. pinnatifida leaves from 4 producing areas. Five known components were identified， including chlorogenic acid （peak 1）， glucosylvitexin （peak 3）， vitexin rhamnoside （peak 4）， hypericin （peak 7） and isoquercetin （peak 8）； their similarities ranged from 0.871 to 0.998. Average relative correlations of samples from Chengde of Hebei Province， Huludao of Liaoning Province， Yuncheng of Shanxi Province and Linyi of Shandong Province were 0.538， 0.528， 0.462 and 0.435， respectively. CA and PCA showed that the samples from Chengde of Hebei Province and Huludao of Liaoning Province were roughly classified into one category， while the samples from Linyi of Shandong Province and Yuncheng of Shanxi Province were roughly classified into one category； VIP values of peak 1， 2， 3 and 5 were all greater than 1. By QAMS， the relative correction factors of chlorogenic acid， glucosylvitexin， hypericin and isoquercetin were 0.401， 0.993， 1.670 and 1.615 （RSD＜2%）. Compared with external standard method， except for isoquercetin in the two batches of samples （S39 and S41）， there was no significant difference in the content of each component in other batches of samples （the relative deviations≤ 5%）. CONCLUSIONS The established fingerprint and QAMS method are simple to operate and can be used to evaluate the quality of C. pinnatifida leaves. The sample from Chengde of Hebei Province is relatively good in quality. Chlorogenic acid （peak 1）， glucosylvitexin （peak 3）， and the corresponding components of peaks 2 and 5 may be differential markers affecting the quality of C. pinnatifida leaves.

Quality evaluation of Xin ’an capsules by combination of fingerprint ，multi-component quantitative analysis and chemical pattern recognition analysis / 中国药房

OBJE CTIVE To establish a method for quality evaluation of Xin ’an capsule by combining fingerprint ， multi-component quantitative analysis and chemical pattern recognition analysis. METHODS High performance liquid chromatography（HPLC）combined with Similarity Evaluation System of TCM Chromatogram Fingerprint （2012 edition）were used to establish the fingerprints of 24 batches of Xin ’an capsules and evaluate the similarity. The common peaks were determined. The contents of glucosylvitexin ，rhamnosylvitexin，vitexin，hyperoside and isoquercetin in Xin ’an capsules were determined by the same HPLC method. Taking the common peak area of fingerprint as the variable ，MetaboAnalyst 5.0 tool was used to draw the cluster analysis （CA）heat map. SIMCA 14.1 software was used to perform principle component analysis （PCA）and partial least squares-discriminant analysis （PLS-DA）. RESULTS Twelve common peaks were identified with the similarity greater than 0.97. Six common peaks were identified as chlorogenic acid ，glucosylvitexin，rhamnosylvitexin，vitexin，hyperoside and isoquercetin.The linear range of glucosylvitexin ，rhamnosylvitexin，vitexin， hyperoside and isoquercetin were 2.36-151.35，9.15-585.20， 1.20-76.50， 0.68-43.20， 0.44-27.90 µg/mL（all r＞0.999）.RSDs of precision ，repeatability and stability （24 h）tests were 163.com all less than 2.00％ . The average recoveries were 95.80％（RSD＝0.96％ ，n＝6），102.10％ （RSD＝0.93％ ，n＝6）， 103.26％（RSD＝1.28％，n＝6），103.89％（RSD＝0.73％，n＝6） and 102.09％（RSD＝1.79％，n＝6），respectively. The contents of the five components were 0.988 8-1.559 1，4.336 6-11.220 1， 0.065 1-0.830 5，0.043 8-0.692 5 and 0.023 2-0.427 2 mg/grain，respectively. The results of CA and PCA showed that 24 batches of samples could be divided into three categories ，i.e. S 1-S15，S16-S18 and S 19-S24. PLS-DA showed that variable importance in projection values of the corresponding component of peak 6 and glucosylvitexin （peak 7），rhamnosylvitexin（peak 8），hyperoside （peak 10） and isoquercetin （peak 11） were greater than 1. CONCLUSIONS The established HPLC fingerprint and multi-component quantitative method are simple and feasible. Combined with chemical pattern recognition analysis ，it can be used for the quality control of Xin ’an capsules. Glucosylvitexin ，rhamnosylvitexin and other components may be differentital markers affecting the quality of each batch of samples.

Analysis of clinical features and pathogenic variants in a Chinese pedigree affected with congenital glycosylation disease / 中华医学遗传学杂志

OBJECTIVE@#To explore the clinical features and genetic basis for a Chinese pedigree diagnosed with congenital glycosylation disease (CGD).@*METHODS@#Clinical manifestations of two brothers were analyzed. Whole exome sequencing was carried out for the sib pair. Suspected variants were verified by Sanger sequencing.@*RESULTS@#Both the proband and her younger brother were found to carry compound heterozygous variants of the PMM2 gene, which included a known pathogenic mutation of c.395T>C (p.I132T) and a previously unreported c.448-1(delAG) in the 5' end of exon 6 of the gene.@*CONCLUSION@#The compound heterozygous variants of the PMM2 gene probably underlay the CGD in the sib pair.

The clinical significance of the transforming growth factor-β receptor I expression in na?ve CD4 + T cells from patients with systemic lupus erythematosus / 中华风湿病学杂志

Objective:To investigate the clinical significance of the transforming growth factor-β receptor I (TGF-βRⅠ) expression in na?ve CD4 + T cells from patients with systemic lupus erythematosus (SLE). Methods:Na?ve CD4 + T cells were purified using magnetic microbeads from peripheral blood mononuclear cells of SLE patients and healthy controls. Real-time quantitative PCR was used to detect TGF-βRⅠ mRNA level, and flow cytometry was used to detect the percentage of CD69 +CD4 + T cells. Data were analyzed by t test and Pearson correlation analysis. Results:The level of TGF-βR Ⅰ mRNA in na?ve CD4 + T cells from SLE patients was significantly lower than that in healthy controls [(0.674±0.873) vs (1.445±1.112), t=2.301, P<0.05]. The TGF-βR Ⅰ mRNA level was negatively correlated with systemic lupus erythematosus disease activity index (SLEDAI) ( r=-0.376, P<0.05), erythrocyte sedimentation rate (ESR) ( r=-0.376, P<0.05), serum creatinine ( r=-0.323, P<0.05) and 24 h urine protein ( r=-0.331, P<0.05), and positively correlated with serum com-plement C3 ( r=0.528, P<0.01). The level of TGF-βRⅠ mRNA level in na?ve CD4 + T cells in SLE patients with renal involvement was lower than that in SLE patients without renal involvement [(0.525±0.536) vs (1.071±1.007), t=2.198, P<0.05]. The TGF-βR Ⅰ mRNA level in the na?ve CD4 + T cells in anti-dsDNA antibody positive group was lower than that in the anti-dsDNA antibody negative group [(0.344±0.315) vs (0.958±1.076), t=2.277, P<0.05]. The expression of TGF-βRⅠ mRNA in na?ve CD4 + T cells from SLE patients was reduced after 24 h stimulation with anti-CD3/CD28 beads [(0.047±0.013) vs (1.008±0.129), t=14.38, P<0.01], which was partially reversed by dexamethasone treatment [(0.240±0.042) vs (0.047±0.013), t=7.845, P<0.01]. Meanwhile, dexamethasone significantly decreased the expression of CD69 in CD4 + T cells [(15.0±2.1)% vs(34.9±2.0)%, t=32.57, P<0.01]. Conclusion:The abnormally low expression of TGF-βRⅠ in na?ve CD4 + T cells may be involved in the pathogenesis of SLE. Glucocorticoid treatment can upregulate the expression of TGF-βRI and inhibit the activation of T cells, This suggests suggesting that TGF-βRⅠ may be a potential target for SLE treatment.

Study on the Quality of Scutellaria baicalensis with Different Harvesting Periods by IR-TL and HPLC-MS / 中国药房

OBJECTIVE： To study effective component kinds and content difference of Scutellaria baicalensis with different harvesting periods collected in spring and autumn， and to determine the optimal harvesting time. METHODS： IR-IL was used to analyze the constitutions change of the components in S. baicalensis collected in different harvesting periods （annual， biennial and triennial， in spring and autumn， respectively）. HPLC-MS method was used to analyze the contents change of 6 flavonoids （qroxylin A， chrysin， wogonin， wogonoside， baicalin， baicalein）. RESULTS： Results of IR-TL analysis showed that S. baicalensis contained different components such as flavonoids， glycosides， saccharides， easters in different growth stages， among which the contents of glycosides in biennial S. baicalensis were the highest with more stable quality. The results of content determination of 6 flavonoids showed that different harvesting periods had a great influence on the contents of the above indicators. The overall content of biennial samples collected in spring was higher than that of autumn samples collected in the same year. CONCLUSIONS： This study provides a reliable analytical method for the quality control of S. baicalensis； spring harvest in two-year growth period may be the best harvesting time for S. baicalensis.

Establishment and characterization of a rat model of hypertension with hyperlipidemia / 中国比较医学杂志

Objective To develop an ideal hypertension combined hyperlipidemia(HP/HL)rat model by feeding spontaneously hypertensive rats(SHRs)with high fat diet, and to evaluate the pathological changes in target organs including heart and kidney. Methods Twenty 3-week old male SHRs were randomly divided into two groups: normal fat control group(SHR-NC)and high fat group(SHR-HF). Moreover,ten 3-week old male Wistar-Kyoto rats(WKY)were taken as the model control group(WKY-NC). The rats in SHR-HF group were fed with high-fat diet to induce HP/HL, while rats of WKY-NC and SHR-NC groups were fed with normal diet. The systolic blood pressure(SBP)and body weight were measured every week. At the end of the experiment, the rats were sacrificed to take serum samples for blood lipid analysis including high density lipoprotein(HDL-C), low density lipoprotein(LDL-C), total cholesterol(TC)and triglyceride(TG). Heart and kidney tissue samples were collected to examine the pathological changes using HE and Masson staining. Results Compared with the SHR-NC group, the SHRs fed with high-fat diet for 23 weeks presented significant increase of blood pressure and TC, TG, LDL-C, and decrease of HDL-C. The HP/HL rat model showed pathological changes in the HP/HL target organs, heart and kidney. Renal tissues were severely damaged and showed a large area of fibrosis. Besides, left ventricular hypertrophy and myocardial fibrosis were also observed. Conclusions A HP/HL rat model is successfully constructed by feeding SHRs with high-fat diet for 23 weeks. Most importantly,this model exhibits progressive renal and cardiac alterations, similar to those of patients with HP/HL. This HP/HL rat model may become widely used for evaluation of HP/HL therapeutic drugs.

The prognostic value of 18F-FDG PET/CT in patients with extranodal natural killer/T-cell lymphoma / 中国肿瘤临床

Extranodal natural killer/T-cell lymphoma(ENKTL)is an Epstein-Barr virus-associated aggressive non-Hodgkin's lymphoma (NHL).It is prevalent in Asia and South America with an aggressive clinical course and a dismal prognosis.Currently,the prognostic models that are commonly used for ENKTL include the International Prognostic Index(IPI),Korean Prognostic Index(KPI),Prognostic In-dex of Natural Killer Cell Lymphoma(PINK),and Prognostic Index of Natural Killer Cell Lymphoma with Epstein-Barr virus DNA(PINK-E), all of which have some limitations.Positron emission tomography/computed tomography(PET/CT)is a new imaging technique combin-ing anatomical and functional imaging,with the advantages of high sensitivity and specificity,which is essential for diagnosis,staging, efficacy evaluation,and prognosis prediction of various malignant lymphomas.However,the prognostic value of PET/CT in ENKTL re-mains controversial.Therefore,in this article,we review relevant studies on the prognostic value of pretreatment,interim,and end-of-treatment PET/CT in patients with ENKTL.

Use of cone-beam computed tomographyto demonstrate morphology of maxillary sinus and association of-maxillary sinus with maxillary posterior teeth / 实用医学杂志

Objective To study the morphological variations of maxillary sinus and to provide anatomical basis for maxillary posterior teeth implantation and extraction usingcone-beam computed tomography (CBCT) scan-ning. Methods CBCT images from 403 patients were reviewed. The minimal distance from sinus floor to alveolar ridge wasmeasured and the number of patients with clinical features such as having sinus septum , mucosal thicken-ing and posterior teeth roots extend into maxillary sinus were counted. Results The mean value of the minimal dis-tance from maxillary sinus floor to alveolar ridge was about 8.3 mm, there was no significant statistical difference between sex, but there was a significant statisticalsignificant difference between the left sides and the right sides. Occurrences of sinus septum, mucosal thickening and roots into sinus were 98.6%, 31% (in which 56.3% of mu-cosal thickening were related to apical periodontitis) and 47%. there was no significantstatisticaldifference between the two sides in males females. Conclusions The morphological variations of maxillary sinus are significant. Chron-ic inflammation in maxillary sinuis is closely related to the position of maxillary posterior teeth roots and periapical lesions.

Effects of artesunate on interstitial pneumonia and submaxillaritis in MRL/lpr mice / 国际中医中药杂志

Objective To investigate the effects of artesunate (ART) on interstitial pneumonia and sialadenitis in MRL/lpr mice.Methods A total of 18 MRL/lpr mice were randomly allocated to a hydroxychloroquine sulfate (HCQ) group,a ART group and a control group.At the age of 18 weeks,the mice in the HCQ group and ART group were given HCQ 150 mg/kg daily and ART 50 mg/kg daily for 12 weeks,respectively.The histopathological changes of pneumonitis and submaxillaritis were assessed by hematoxylin and eosin staining.The levels of monocyte chemoattractant protein-1 (MCP-1) in the serum and urine were measured by the enzyme-linked immunosorbent assay.Results At the age of 30 weeks,the index of peribronchiolar lesion (1.62 ± 0.19,1.52 ± 0.30 vs.1.95 ± 0.34;all P＜0.05),the index of perivascular lesion (1.23 ± 0.18,1.28 ± 0.12 vs.1.57 ± 0.33;all P＜0.05),the alveolar lesions index (1.35 ± 0.16,1.05 ± 0.15 vs.1.72 ± 0.34;all P＜0.05) and the submaxillaritis index (1.48 ± 0.22,1.43 ± 0.15 vs.1.84 ± 0.34;all P＜0.05) in the HCQ group and the ART group were significantly decreased than those in the control group.The MCP-1 levels in the serum (1 103.02 ± 185.56 pg/ml,1 072.37 ± 242.43 pg/ml vs.1 490.67 ± 329.43 pg/ml;all P＜0.05) and urine (189.16 ± 70.85 pg/ml,198.79 ± 113.47 pg/ml vs.446.79 ± 192.31 pg/ml;all P＜0.05) in the HCQ group and the ART group were significantly lower than those in the control group.Conclusion ART can decrease the MCP-1 level,and ameliorate interstitial pneumonitis and sialadenitis in MRL/lpr mice.

Simultaneous Determination of Six Effective Components in Crataegus pinnatifida by Quantitative Analysis of Multi-components by Single Marker / 中国药房

OBJECTIVE:To establish a method for the simultaneous determination of chlorogenic acid,vitexin glucoside,vi-texin rhamnoside,vitexin,rutin and hyperoside in Crataegus pinnatifida. METHODS:With reference peak of vitexin glucoside, HPLC was conducted to calculate the relative correction factor(RCF)of chlorogenic acid,vitexin glucoside,vitexin rhamnoside, vitexin,rutin and hyperoside,then the contents of above-mentioned 5 components in C. pinnatifida were calculated. The column was Agilent ZORBAX SB C18 with mobile phase of 0.1% formic acid-acetonitrile-tetrahydrofuran (gradient elution) at a flow rate of 1.0 ml/min,the detection wavelength was 350 nm,column temperature was 30 ℃,and the injection volume was 10 μl. RE-SULTS:The linear range was 12.50-400.0 μg for chlorogenic acid(r=0.999 8),25.00-800.0 μg for vitexin glucoside(r=0.999 9), 31.25-1 000.0 μg for vitexin rhamnoside(r=0.999 9),6.470-260.0 μg for vitexin(r=0.999 9),2.50-80.0 μg for rutin(r=0.999 8) and 9.375-300.0 μg for hyperoside(r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 2.0%;re-coveries were 99.2%-103.9%(RSD=1.6%,n=6),97.9%-100.8%(RSD=1.2%,n=6),99.2%-100.8%(RSD=0.5%,n=6), 97.3%-101.3%(RSD=1.5%,n=6),98.0%-103.0%(RSD=1.9%,n=6)and 95.5%-101.5%(RSD=2.2%,n=6). RCFs of vitex-in glucoside with chlorogenic acid,vitexin rhamnoside,vitexin,rutin and hyperoside were 1.119,1.009,0.706,1.063 and 0.830, respectively. CONCLUSIONS:The method is simple with good precision,stability and reproducibility,and it can be sued for the simultaneous determination of 6 components in C. pinnatifida.

The clinical significance of tumor budding in predicting lymph node metastasis of T1 colorectal cancer / 中国癌症杂志

Background and purpose:Tumor budding is a poor prognostic factor in colorectal cancer. In this study, we studied the tumor budding by counting the actual number in 10 high power fields and evaluated itsclinical application in predicting lymph node metastasis of T1 colorectal cancer.Methods:Tissue specimens from 307 patients with histologically conifrmed T1 colorectal cancer were enrolled. The clinicopathological characteristics including tumor budding were evaluated for their predictive value in lymph node metastasis. A formula was created to calculate the risk score for prediction of lymph node metastasis which was validated by 14 new cases.Results:In the multivariate analysis, it showed that tumor grade, lymphovascular invasion and the number of tumor budding were signiifcantly associated with lymph node metastasis. The probability of lymph node metastasis was calculated using the following equations:Z=1.571×(lymphovascular state: invasion, 1; no invasion, 0)+2.661×(tumor grade: high grade, 1; low grade, 0)+0.024×(budding counts)-3.885; Probability=1/1+e-Z. The high scores were correlated with the lymph node metastasis in the validations.Conclusion:We can accurately assess the risk of lymph node metastasis by counting the number of tumor budding in 10 high power fields. Therefore tumor budding could potentially assist treatment decision making in T1 colorectal cancer patients with high-risk lymph node metastasis.

Determination of Six Kinds of Flavonoids in Scutellaria baicalensis Georgi with Different Determination of Six Kinds of Flavonoids in Scutellaria baicalensis Georgi with Different Storage Time by LC-MS / 医药导报

Objective To quantify the contents of baicalin, wogonoside, baicalein, wogonin, chrysim and oroxylinA in the Scutellaria baicalensis Georgi with different storage time by liquid chromatography-mass spectrometry( LC-MS) and determine the stability of Scutellaria baicalensis Georgi. Methods The LC system consisted of an Agilent Zorbax XDB-C18 column (2. 1 mm×150 mm,3. 5 μm) with acetonitrile and water (0. 1% formic acid) in linear gradient condition. The flow rate was 0. 2 mL·min-1 and the wavelength of detection was 276 nm. The mass spectrometer was operated under the positive ion mode with the ESI source. The precursor-to-product ion pair was 447/271 for baicalin,461/285 for wogonoside,271/123 for baicalein, 285/270 for wogonin, 255/153 for chrysim and 285/270 for oroxylinA, respectively. Results The contents of baicalin, wogonoside, baicalein, wogonin, chrysim and oroxylin A had a linear relationship with their peak areas within the ranges of 8. 86-177. 2, 2. 33-46. 5, 2. 6-52. 0, 0. 52-10. 4, 0. 04-0. 8 and 0. 04-0. 8 μg · mL-1 , respectively. The contents of baicalin, wogonoside, baicalein, wogonin, chrysim, and oroxylin A were overall decreased over the first 3 months of storage,but remained relatively stable between 0 and 18 months. Conclusion The results provides basis for the establishment of quality standard.

Research on proportional control system of prosthetic hand based on FMG signals / 生物医学工程学杂志

The control of prosthetic hand is always a focus in prosthesis research. For solving current problems of controlling signals of skin surface electrical signals, we applied force myography (FMG) signals in prosthetic control of this system. The control system based on FMG signals were designed, containing signal acquisition and pre-processing, prosthetic control, motor driving and so on. Two-freedom artificial hand with proportional control was proposed through acquiring two-channel FMG signals from the amputee stump. The proportional control of prosthetic hand was achieved according to the average of FMG amplitude. The results showed that the control system had a great potential to control artificial hand and to realize speed adjustment effectively. Besides, the Virtual instrument software LabVIEW is adopted to establish the FMG signal collection and calibration of experiment system.

Clinical analysis of allogenic bone marrow derived mesenchymal stem cells transplantation for 11 patients with systemic lupus erythematosus / 中华风湿病学杂志

Objeefive To explore the clinical efficacy and safety of allogenic bone marrow derived mesenchymal stem cells transplantation(MSCT) in patients with refractory systemic lupus erythematosus(SLE).Methods Eleven patients with refractory SLE(nine females and two males aged from 16～41 years(mean 25±8).were entailed in the study.The infcIrmed consents which were approved by the Ethics Committee of the Affiliated Drum Tower Hospital of Naniing University Medical School were obtained from all participants.Bone marrow of healthy donors were obtained and the mesenchymal stem cells(MSC)were expanded in vitro.Each patient was infused MSC 1×106/kg body weight intravenously.Before MSCT.all patients were administrated with cvclophosphamide (CTX)800～1800 mg divided by two to three days.The clinical manifestations and laboratory tests were compared before and after MSCT.Results The eleven patients were followed up for one to thirteen months after MSCT.A11 patients did not develop transplantation related complications.The systemic lupus erythematosus disease activity index (SLEDAI)score decreased from 1 1.7±5.1 to 5.6±3.4 one month after MSCT(n=11,P＜0.01).Ufine protein excretion decreased from(1989+842)mg/24 h to(1118±700)mg/24 h one month after MSCT(n=10,P=0.02).Five patients were followed up for six months and their urine protein excretion decreased significantly [(522±151)mg/24 h vs (2478±797)rag/24 h.n=5.P＜0.01j.The serum albumin level of 5 patients with hypoalbuminemia increased gradually one month after MSCT [(28±6)g/L vs (32±7)g/L,n=5,P＜0.05].Serum complement C3 level increased from(0.50±0.12) g/L to(0.75±0.10)g/L (n=9.P＜0.01) and their anti-nuclear antibody (ANA) titer decreased one month after MSCT.In two patients with chronic renal failure.the serum creatinine decreased gradually.Conclusion Allogenic MSCT is an effective and safe approach for the treatment of refractory SLE.However.extensive follow-up study is needed for long-term benefit evaluation.

Abnormality of multilineage difierentiation potential of bone marrow-derived mesenchymal stem cells in patients with systemic lupus erythematosus / 中华风湿病学杂志

Objective To investigate the muhilineage differentiation potential of bone marrow-derived mesenchymM stem eels (MSCs) in patients with systemic lupus erythematosus (SLE).Methods Density gradient centrifugation and plastic adherence methods were used for isolation of marrow-derived MSCs.Then tIIeir differentiation potentiality to lipoblasts and osteoblasts waft tested.MSCs loading on hydroxyapatite were elnbedded in the nude mouse's subcutaneous tissues.Eight weeks later.osteogenesis was evaluated by HE staining.PPA Rγ2,LPL,Runx2/CBFA1,osteocalcin gene expression in MSCs after differentiation were examined by RT-PCR.Results The positive rates of lipoblasts stained by oil red O and optical density in SLE were decreased than in the control group[(35±7)% vs (80±5)%] (0.14±0.04 vs 0.27±0.04),and the positive rates of osteoblasts stained by Alizarin Red S in SLE were decreased than those in the control group [(35±4)% vs (45±4)%].Osteoblast differentiation in the SLE group was less than that of the contro]group.The mRNA expression of LPL (0.369±0.020 vs 0.481±0.038).Runx2/CBFA1 (0.371±0.000 vs 0.563±0.069).osteoealcin (0.819±0.023 vs 0.962±0.049) of MSCs after difierentiation in the SLE group was decreased than that of the control group.There was no significant difference in the expression of PPARγ2 mRNA between SLE and controI group (0.421±0.052 vs 0.441±0.012).Conelusion MSCs from SLE have abnormalities in osteogenie and adipogenic differentiation potential.

The effect of hydroxychloroquine treatment on MRL/lpr lupus mice / 中华风湿病学杂志

Objective To investigate the therapeutic effects and mechanisms of hydroxychloroquine (HCQ) in the MRL/lpr mice. Methods MRL/lpr mice were divided into HCQ, the artesunate (ART) and proteinuria was detected with Coomassi Brilliant blue method. Enzyme linked immunosorbent assay (ELISA) was used to measure the anti-doubM-stranded DNA (ds-DNA) antibody. Renal tissue sections were dyed By PAS methods. The percentage of CD4+ Foxp3+ T cells in the spleen and lymph nodes were detected by flow 2.0) mg groups were decreased than in the control group (4.8±3.2) mg (P<0.05). And it was also lower in the HCQ (2.8±1.1) mg and ART (2.4±1.9) mg group than in the control group (6.4±1.9) mg (P<0.01) at 30 in the control group (37.1±1.0) g (P<0.01), while serum creatinine decreased significantly (7.8±4.0) μmol/L than in the control group (12.5±2.3) μmol/L (P<0.05), and the serum anti ds-DNA antibodies levels (3047±renal damage in the HCQ group and in the ART group was Both significantly improved than that in the entages of CD4+ Foxp3+ T cells in spleen when compared with the control group (1.5±0.5)% (P<0.05). The mice in the HCQ group (0.68±0.33)% and in the ART group (0.97±0.28)% had higher percentages of CD4+ Foxp3+ T cells in lymph nodes as compared with control group (2.15±0.72)%(P<0.01). Conclusion HCQ is effective in treating MRL/lpr lupus mice. It can improve the pathologic lesions of lupus nephritis, reduce proteinuria and antibody production. Both HCQ and ART can up-regulate the percentage of CD4+ Foxp3+ T cells in spleen of MRL/lpr mice.

The effect of umbilical cord mesenchymal stem cells transplantation for the MRL/Ipr mice / 中华风湿病学杂志

Objective To investigate the efficacy of umbilical cord mesenchymal stem cells (UC-MSCs) transplantation in the treatment of the MRL/lpr mice. Methods Twenty four 18-week-old MRL/lpr female mice were divided into 3 groups:group 1 (G1) were transplanted with 1×106 UC- MSCs through caudal vein, group 2 (G2) were transplanted with 1×106 UC- MSCs three times and group 3 (G3) were treated with 0.5 ml normal saline as controls. Enzyme linked immunosorbent assay (ELISA) was used to measure the levels of serum anti-dsDNA antibodies. Twenty-four hours proteinuria and body weight were assessed every two weeks. The histopathology changes of the kidneys and lungs were observed. Results ① At the 25th weeks, the 24 hours proteinuria in group G1 (2.3±1.9) mg and G2 (1.8±1.4) mg was decreased than that in the control group (3.8±2.1) mg (P<0.05), and at the 27th weeks, that of groups G1 (2.5±1.5) mg and G2 (1.9±1.2) mg was also significantly decreased than in the control group (5.4±2.4) mg (P<0.01); ② From the 24th week, the body weight of groups G1 and G2 increased significantly than that of the control group (P< 0.05). At week 29, serum creatinine decreased significantly in both groups G1 (7.2±3.2) μmol/L and G2 (6.2±2.8) μmol/L than in the control group (12.5±2.3 ) μmol/L (P<0.05); ③One week after transplantation, the levels of anti-dsDNA antibodies in group G1 (46±11)×102 U/ml and G2(49×43)×102 U/ml were bothsignificantly decreased than those of the control groups (99±42)×102 U/ml (P<0.05) and the difference between group G2 (36±15)×102 U/ml and the controls (68±32)×102 U/ml was statistically significant; ④The nephron crescent formation in group G1 (0.12±0.07) and G2 (0.08±0.02) was significantly lower that of the control group (0.20±0.06) (P<0.05) and that of group G2 was significantly less that of froup G1 (P<0.05); ⑤ The interstitial pneumonitis was singnificantly milder in group G1 than group G2. Conclusions UC- MSCs is very effective in treating MRL/lpr mice. It is safe and free of rejection reactions.

Regulation of mesenchymal stem cells on CD4+ Foxp3+ T cells in systemic lupus erythematosus / 中华风湿病学杂志

Objective To investigate the in vivo or in vitro immune regulatory effects of allogeneic bone-marrow mesenchymal stem ceils (MSC) and human umbilical cord MSC on CD4+ Foxp3+ T cells in the peripheral blood of patients with systemic lupus erythematosus (SLE) and in the spleen of MRL/Ipr mice. Methods Human MSC were isolated and expanded from bone marrow cells of healthy donors and infused into five SLE patients. The percentages of CD4+ Foxp3+ T cells in peripheral blood were detected by flow cytometry. Human peripheral blood mononuclear cells (PBMC) were prepared by centrifugation on a Ficoll Hypaque density gradient. The MSC and PBMC from unrelated donors (MSC:PBMC =1:1,1:10,1:50) were added into 24-well plates. After 72h of co-culture, the percentages of CD4+ Foxp3+ T cells were detected by flow cytome- try. Twenty four 18-week-old MRL/Ipr female mice were divided into 3 groups and were injected with umbilical cord MSC (1×106 cells for one time, 1×106 cells for three times and 0.5 ml sodium chloride as control respectively). The percentages of CD4+ Foxp3+ T cells in spleen and lymphoid nodes were detected by flow cytometry. Results The percentages of blood CD4+ Foxp3+ T cells at one week (4.8± 1.6)% and at three months (6.0±2.6)% post MSC transplantation for patients with SLE were both higher than that before transplantation (2.1±1.2)% (n=5,P<0.05). The co-culture of normal bone marrow MSC with PBMC from SLE patients resulted in a statistically significant increase of CD+ Foxp3+ T cells percentage in PBMC on a dose dependent manner (P<0.05). The percentages of CD4+ Foxp3+ T cells of PBMC from SLE patients co-cultured with lupus MSC were lower than that of normal MSC (P<0.05). The cultured supematant of normal MSC also upregulated the percentages of CD4+ Foxp3+ T cells of PBMC from SLE patients (P<0.05). The MRL/lpr mice that had been injected umbilical cord MSC for one time and three times had higher percentages of CD4+ Foxp3+T cells in the spleen but lower in the lymphoid nodes as compared with controls (P<0.01), but without statistical significant difference. Conclusion Allogeneic or heterogeneie MSC transplantation upregulates the percentages of CD4+ Foxp3+ T cells in SLE patients or in MRL/Ipr mice. Upregnlation of Treg population may be one of the mechanisms of MSC transplantation that is effective for SLE treatment.

Characterization of the ultrastructure and cytoskeleton of bone marrow-derived mesenchymal stem cells in patients with systemic lupus erythematosus / 中华风湿病学杂志

Objective To explore ultrastructure and cytoskeleton characteristics of bone marrow-deftved mesenchymal stem cells (MSCs) in patients with systemic lupus erythematosus (SLE).Methods MSCs were isolated from bone marrow of 2 SLE patients and 2 healthy controls.Their ultrastrnctures were examined by transmission electron microscope (TEM).The expression pattern of actin and vinculin was assessed by laser confocal microscopy (LCM).Results MSCs in patients with SLE presented with signs of ageing and lots of autophagosome could be found in most of the cells.F-actin was aggregated and condensed at the:border of cytoplasm.Vinculin was arranged disorderly and condensed in the cytoplasm.Conclusion The change of uhrastructure and cytoskeleton patterns of bone marrow derived mesenchymal cells of SLE patients may play an important role in the abnormal proliferation of these cells in vitro.