Objective To explore the feasibility of differentiation into neural cells from bone marrow stromal cells(BMSCs) in vitro by breviscapine,and to offer reference for the applying of BMSCs. Methods The bone marrow stromal cells of SD rat were separated and purified by passsage culture.After phenotype characterization,the 4th passage of BMSCs was exposed to breviscapine. Differentiation was observed under phase contrast microscope every 6 hours and the cells were stained immunocytochemically with neuronspecific enolase(NSE)and glial fibrillary acidic protein(GFAP).The induced cells activity was detected by MTT. NSE and GFAP were determined by flow cytometry and RT-PCR. Results The BMSCs were CD44~+,CD54~+,CD34~-. After 18 hours of induction, cytoplasm of BMSCs partly contracted with protruding;The immunocytochemical staining was performed on cells after induction for 24 hours,the rates of NSE and GFAP staining positive were(48.7±3.4)% and(56.8±4.2)%.By using flow cytometer, expressions of NSE and GFAP showed much higher in induced group than that in control group. By using RT-PCR, expressions of NSE and GFAP were positive in induced group. Conclusion Breviscapine could induce adult rat BMSCs to differentiate into neuron and glial cells.

Objective To investigate the time course of cerebral infarction with diffusion kurtosis imaging-related parameters.Methods According to the time interval from symptom onset to MRI examination, 114 cases of cerebral infarction were divided into five groups:8 cases of hyperacute phase(less than 6 hours), 14 cases of acute(>6-24 h), 60 cases of early subacute(>24 h-7 d), 20 cases of late subacute(>7-14 d), and 12 cases of chronic phase ( >14 d-2 months).They underwent routine diffusion weighted imaging (DWI) and diffusion kurtosis imaging (DKI) scanning, and apparent diffusion coefficient ( ADC) and DKI-derived parameters were obtained from them.The derived diffusion parameters were compared among different phases in the patients , and the percent of changes in the infarcted regions were calculated.Paired t-test was used to compare the difference of each parameter between the infarcted region and contralateral normal region , and their correlation with time interval was tested using Pearson correlation analysis.Results Except for chronic phase , mean kurtosis ( MK) , axial kurtosis ( K∥) , radial kurtosis (K⊥)map showed uneven high signal in the infarcted regions , while mean diffusion(MD), axial diffusion(D∥), radial diffusion(D⊥) showed uniform low signal.MK values in the infarcted regions of hyperacute, acute, early subacute and late subacute phase (1.331 ±0.357,1.578 ±0.453,1.519 ±0.455, 1.403 ±0.275 ) increased significantly , compared with the contralateral normal mirror regions ( 0.850 ± 0.236,0.827 ±0.194,0.865 ±0.144,0.939 ±0.212) (t values were 5.242,6.907,12.416,5.629, respectively.P values were all less than 0.01 ).MK, K∥, K⊥ achieved a peak in the acute and early subacute phase , and showed more amplitude than the decrease of MD , D∥, D⊥.Then they gradually reduced, and tended to normalize.MK, MD, ADC had a significant correlation with the time onset of cerebral infarction ( r was 0.354, 0.747, 0.723, respectively, P values were all less than 0.05 ).Conclusion Diffusion kurtosis imaging can provide more diffusion information than conventional DWI , which can better reflect the microstructure changes in tissue.

To obtain a vaccine to defend from dormancy Mycobacterium tuberculosis, we constructed the recombinant Bacilli Calmette-Guérin (BCG) vaccine with Rv3133c encoding dormancy-correlated transcriptional regulatory protein DosR in Mycobacterium tuberculosis as a target gene, and evaluated its immunogenicity in BALB/c mice. In this study, we constructed the recombinant plasmids of rpMV361-Rv3133c using gene colon technology. We then transformed BCG strains with above-mentioned plasmids to obtain recombinant vaccine of rBCG-Rv3133c. We used the rBCG strains successfully constructed to vaccinate in BALB/c mice. 30d and 180d after immunization, the specific antibody titers were determined to investigate humoral responses induced by recombinant vaccine. We detected changes of splenocyte subsets of CD4+T, CD8+ T cells and cytokine of IFN-gamma secreted by splenocytes for evaluation of cellular immune responses. The results showed that the rBCG-Rv3133c was able to induce higher levels of antibody titer, stronger proliferative responses and higher IFN-gamma production comparing with BCG vaccine. The results also suggested that this recombinant vaccine was a more efficacious tuberculosis vaccine for further study.
Animals ; Antibodies, Bacterial ; blood ; Antigens, Bacterial ; genetics ; immunology ; BCG Vaccine ; immunology ; Bacterial Proteins ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Interferon-gamma ; immunology ; Mice ; Mice, Inbred BALB C ; Mycobacterium tuberculosis ; immunology ; Protein Kinases ; genetics ; immunology ; Recombinant Proteins ; immunology ; T-Lymphocyte Subsets ; immunology ; Tuberculosis ; prevention & control ; Vaccination ; Vaccines, Synthetic ; immunology

Twenty-one rabbits were randomly divided into three groups: experimental group-filling vein with two neural segments being 0.3cm long to bridge 4cm defect of common peroneal nerve, control group-bridging vein directly to the 4cm defect of common peroneal nerve, auto-nerve-grafting group-cutting off a segment (4cm long) from common peroneal nerve and grafting it inversely.After 25 weeks, morphological, electrophysiological and histological examinations were undertaken, which revealed that the experimental group was most similar to the auto-nerve-grafting group in recovery of motion of the limbs, action potential of muscle, nerve conductive velocity, and regenerating density of nerve fibers and axons. It was a failure in control group. Satisfactory results were also achieved in 2 patients with defect of ulnar nerve treated by filling vein with two neural segments. It suggests that our method is feasible.

Objective To evaluate the role of Toll-like receptor 4 (TLR4) in non-medullary andmedullary cells in lung ischemia-reperfusion (I/R) injury in mice.Methods Ten healthy male TLR4+/+ in non-medullary cells/TLR4+/+ in medullary cells (WT/WT) mice,10 TLR4-/-in non-medullary cells/ TLR4-/-in medullary cells (KO/KO) homozygote mice,10 TLR4+/+ in non-medullary cells/TLR4-/-in medullary cells (WT/KO) mice,and 10 TLR4-/-in non-medullary cells/TLR4+/+ in medullary cells (KO/WT) heterozygote mice,aged 6-8 weeks,weighing 20-25 g,were used in the study.Lung I/R was induced by occlusion of the left hilum for 60 min followed by 240 min of reperfusion in anesthetized mice.Blood samples were obtained from the femoral artery at 240 min of reperfusion for blood gas analysis,and the oxygenation index (PaO2/FiO2) was calculated.The animals were then sacrificed and lung tissues were immediately removed for determination of wet/dry weight ratio,myeloperoxidase activity and contents of tumor necrosis factor-alpha,interleukin-1beta (IL-1β) and IL-6 (by enzyme-linked immunosorbent assay) and for microscopic examination of the pathological changes of lungs which were scored.Results Compared with WT/WT mice,the oxygenation index was significantly increased in sequence,and lung injury scores,wet/dry weight ratio,myeloperoxidase activity and contents of tumor necrosis factor-alpha,IL-1β and IL-6 were decreased in sequence in WT/KO,KO/WT and KO/KO mice (P＜0.05).Conclusion TLR4 in non-medullary cells plays a rnore important role in lung I/R injury than that in medullary cells of mice.

Colon cancer is one of the most common malignant tumors worldwide.When the conventional treatment methods provide survival benefits for patients,their side effects and limitations cannot be ignored.Many traditional Chinese medicines has attracted much attention from scholars at home and abroad because of its anti-tumor advantages of multi-target,multi-pathway and less toxic side effects.Many Chinese medicines and the active components extracted from them,represented by flavonoids,phenols and alkaloids,have played an effective role in anti-tumor.There mechanism is related to inhibiting tumor cell proliferation,inducing apoptosis,regulate autophagy,inhibiting tumor cell invasion and metastasis,inhibiting tumor angiogenesis.In this paper,the mechanism of inhibiting colon cancer by effective components of traditional Chinese medicine in recent years was reviewed in order to provide reference for the clinical treatment of colon cancer.

Objective To explore the efficacy of quantitative susceptibility mapping (QSM) in the assessment of osteoporosis and the impact factors on the QSM values.Methods A total of 105 volunteers (35 males and 70 females) were recruited in this study.The height,weight,waistline and hipline were measured,and the body mass index was calculated.All the subjects underwent MRI-based QSM and quantitative computed tomography (QCT).The measurement of QSM and QCT values was performed on L3 vertebrae body.According to QCT value,the subjects were divided into three groups (normal,osteopenia and osteoporosis).According to age,the subjects were divided into group I (21-30 years old),group 2 (31-40 years old),group 3 (41-50 years old),group 4 (51-60 years old),and group 5 (＞60 years old).Differences among all groups were compared using one-way ANOVA or Kruskal-Wallis.Results According to QCT value,54 subjects were normal,22 osteopenic and 29 osteoporotic.The QSM value for the subjects with osteoporosis [148.60(109.42,188.81)ppb] was significantly higher than that of normal (P＜0.001)and the osteopenia (P＜0.001).The QSM value for the subjects with osteopenia was significantly higher than the normal (P＜0.001).The coefficient of QSM and BMD was-0.749 (P＜0.001).Multiple linear regression showed age was the independent influence factor for QSM value (r=0.72,P＜0.001),whereas the gender,BMI,waistline and hipline showed no significant difference (P＞0.05).With the increasing of age,the QSM value showed a gradual increasing trend.And there were significant differences of QSM values among the different age groups (P＜0.001).The QSM value of 138.98 (100.37,183.84)ppb for group 5 (＞60 years old) was significantly higher than that of group 1,group 2,and group 3 (P＜0.001).There is no difference between group 5 and group 4 (P＞0.05).The QSM value of 96.62(28.62,143.99)ppb for group 4 (51-60 years old) was significantly higher than that of group 1 and group 2 (P＜0.001).And there was no difference between group 4 and group 3 (P＞0.05).The QSM value of group 1,group 2,and group 3 showed no significant difference (P＞0.05).Conclusions The QSM of bone is feasible in the assessment of osteoporosis and has the potential to be a biomarker providing new insights into osteoporosis.And age is the critical factor affecting QSM value.

Objective To observe the value of three-dimensional arterial spin labeling (3D-ASL) PWI in evaluating postoperative cerebral perfusion changes in patients with Moyamoya disease.Methods Totally 19 patients of Moyamoya disease confirmed with DSA were enrolled.All the patients received revascularization.Before and after operation,3D-ASL PWI and dynamic susceptibility contrast perfusion weighted imaging (DSC-PWI) were performed.ROI was located in the region with obvious perfusion changes supplied by middle cerebral artery on the operating side.Then the cerebral blood flow (CBF) was measured on 3D-ASL images,and time to peak (TTP) was measured on DSC-PWI images before and after operation.The differences of CBF and TTP before and after operation were compared,as well as the improvement rate of CBF,TTP and clinical symptoms.Results Before and after operation,CBF was (41.40±11.36) ml/(100 g · min) and (54.10±16.69) ml/(100 g · min),respectively,and the difference was statistically significant (t=-4.273,P＜0.01).TTP was (28.66 ± 3.21) s and (26.44 ± 3.93) s,respectively,and the difference was also statistically significant (t =-2.936,P＜0.01).The improvement rate of clinical symptoms was 84.21％ (16/19),of CBF was 78.95％ (15/19) and of TTP was 68.42％ (13/19),the differences of improvement rate had no statistically significant (P=0.625).Conclusion 3D-ASL PWI is noninvasive,no contrast agent need to be used,and can be used to evaluate perfusion changes after operation of revascularization in patients with Moyamoya disease.

Objective:To investigate the correlation between neutrophil/lymphocyte ratio (NLR) combined with low-density lipoprotein cholesterol/high-density lipoprotein cholesterol ratio (LDL-C/HDL-C) and severity of coronary lesions in patients with acute coronary syndrome (ACS).Methods:Patients who were diagnosed with ACS due to chest pain and received emergency coronary angiography in the First Affiliated Hospital of University of Science and Technology of China and the Affiliated Hospital of Anhui Medical University from January 2017 to June 2020 were enrolled in the final analysis. The data of gender, age, body mass index (BMI), past history, emergency blood routine indicators [neutrophil (NEU), lymphocyte (LYM), monocyte (MON), eosinophil (EOS), basophil (BAS), red blood cell (RBC), mean corpuscular volume (MCV), blood red cell distribution width (RDW), mean platelet volume (MPV), platelet volume distribution width (PDW)], blood lipid index [triglyceride (TG), total cholesterol (TC), HDL-C, LDL-C, very low-density lipoprotein cholesterol (VLDL-C)], and coronary angiography were collected. The results of coronary angiography were evaluated by the Gensini score. According to the Gensini score, the patients were divided into the control group (Gensini score = 0, 55 cases) and the study group (Gensini score > 0, 889 cases), and then the patients in the study group were divided into the low-Gensini-score group (Gensini score < 66, 419 cases) and the high-Gensini-score group (Gensini score ≥ 66, 470 cases). The differences in the general baseline data of the four groups were compared, and the correlation between the statistically significant data and the Gensini score was linearly analyzed, and then the combined diagnostic factors (NLR combined with LDL-C/HDL-C ratio) were obtained by Logistic regression analysis. The receiver operator characteristic curve (ROC curve) was used to evaluate the predictive value of NLR combined with LDL-C/HDL-C ratio in predicting the severity of coronary artery lesions in patients with ACS. Finally, multivariate linear regression analysis was used to establish the predictive model between NLR combined with LDL-C/HDL-C ratio and Gensini score.Results:A total of 944 patients were finally included. The differences in gender, age, BMI, hypertension, diabetes, smoking history, NEU, LYM, MON, EOS, RDW, TC, HDL-C, LDL-C, NLR, LDL-C/HDL-C ratio between the control group and the study group were statistically significant. The differences in BMI, hypertension, diabetes, smoking history, NEU, LYM, MON, EOS, TG, TC, HDL-C, LDL-C, NLR and LDL-C/HDL-C ratio between the low-Gensini-score group and the high-Gensini-score group were statistically significant. Linear regression analysis showed that compared with other indicators, the correlation between NLR, LDL-C/HDL-C ratio and Gensini score was stronger in the study group ( r values were 0.634 and 0.663, respectively, both P < 0.05). Binary Logistic regression analysis of the indicators related to Gensini score showed that NEU, LYM, HDL-C and LDL-C were independent risk factors for coronary stenosis in patients with ACS [odds ratio ( OR) were 0.189, 10.309, 13.993, 0.251, 95% confidence intervals (95% CI) were 0.114-0.313, 4.679-22.714, 3.402-57.559, 0.121-0.519, respectively, all P < 0.05]. ROC curve analysis showed that NLR combined with LDL-C/HDL-C ratio had higher predictive value in predicting the severity of coronary lesions in ACS patients [area under the ROC curve (AUC) was 0.952, 95% CI was 0.93-0.969], when the cutoff value was -3.152, the sensitivity was 98.20%, and the specificity was 81.60%. According to the results of multivariate linear regression analysis, the prediction model between NLR, LDL-C/HDL-C ratio and Gensini score was established, and the formula was Gensini score = -7.772+15.675×LDL-C/HDL-C ratio+8.288×NLR ( R2 = 0.862). Conclusion:There is a significant correlation between emergency NLR combined with LDL-C/HDL-C ratio and Gensini score in patients with ACS at admission, which has a certain predictive value for the severity of coronary artery stenosis in patients with ACS, and can be used as a predictor for evaluating the severity of coronary artery disease.

Objective: To investigate the levels of NK cells and their relevant cytokines (IL-10, TGF-β and IFN-γ) in patients with primary immune thrombocytopenia (ITP) . Methods: All samples were obtained from 42 patients (22 newly diagnosed and 20 in remission) and 20 healthy volunteers. The levels of IL-10 and IFN-γ in blood serum were detected by enzyme-linked immunosorbent assay (ELISA) . The percentage of CD3^- CD56⁺ NK cell, CD3^- CD56^bright CD16^- NK cell, CD3^- CD56^dim CD16⁺ NK cell in peripheral blood lymphocyte were detected by flow cytometry. The NK cells were isolated by immunomagnetic microbeads. The mRNA expression levels of IL-10, TGF-β, and IFN-γ in NK cells were detected by real-time fluorescent quantitative PCR. Correlation between the above measured results was analyzed. Results: ① The blood serum level of IFN-γ in newly diagnosed ITP patients [ (653.0±221.6) ng/L] was higher than that in remission ITP patients [ (484.4±219.5) ng/L] and healthy control [ (390.9±253.5) ng/L] (P=0.022, P=0.001) . The blood serum level of IL-10 in newly diagnosed ITP patients was lower than that in healthy control [ (52.09±26.66) ng/L vs (79.44±38.43) ng/L, P=0.007]. ②The percentage of NK cell in newly diagnosed and remission ITP patients [ (9.53±3.93) %, (9.03±3.78) %] were significantly lower than that in healthy control [ (13.72±7.42) %] (P=0.013, P=0.007) . The ratio of CD3^- CD56^bright CD16^- NK cell/total NK cells in newly diagnosed ITP patients was higher than that in healthy control [ (6.85±4.43) % vs (4.05±2.81) %, P=0.032]. The ratio of CD3^-CD56^dim CD16^- NK cell/total NK cells in newly diagnosed ITP patients was lower than that in healthy control [ (93.14±4.43) % vs (95.94±2.81) %, P=0.032]. ③ There was no significant difference in the mRNA expression level of IFN-γ in NK cells of ITP patients and healthy control (all P>0.05) . The mRNA expression levels of IL-10 and TGF-β in NK cells in newly diagnosed ITP patients were significantly higher than that in healthy control (1.82±1.32 vs 1.02±1.03, P=0.023; 2.80±2.31 vs 1.46±1.37, P=0.028) . The ratio of CD3^-CD56^bright CD16^- NK cell/total NK cells was positively correlated with the mRNA expression levels of IL-10, TGF-β in NK cells (r=0.424, P=0.001; r=0.432, P<0.001) . Conclusion NK cells may compensate for the deficiency of the number by enhancing the secretion of negative regulation cytokines, acting as "protective" roles in the disease.