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Chronic hepatitis B virus (HBV) infection is a major cause of viral hepatitis, liver cirrhosis, and hepatocellular carcinoma (HCC). From chronic HBV infection to HCC, most patients go through the stages of chronic hepatitis, liver cirrhosis, and HCC. During this long process, the ongoing integration of HBV DNA into host DNA increases the risk of HCC, and the death and compensatory proliferation of hepatocytes caused by persistent liver inflammation may promote the accumulation of oncogenic mutations and finally lead to the malignant transformation of hepatocytes. Currently, nucleos(t)ide analogues are widely used anti-HBV drugs, which controls infection by inhibiting HBV replication and can thus effectively slow down disease progression and end-stage liver disease; however, anti-HBV therapy often starts late and has a relatively low treatment rate, and there is still a tendency of increase in the incidence rate of HBV-related HCC. Therefore, how to improve current antiviral strategies to further reduce the risk of HBV-related end-stage liver disease including HCC has become a hotspot in clinical practice. This article summarizes the previous studies supporting the expansion of antiviral therapy and suggests that antiviral therapy should be initiated as early as possible to inhibit viral replication and the sequential events of HBV DNA integration and ultimately reduce the risk of HCC in patients with chronic HBV infection.
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Objective: To explore the feasibility and application value of intraoperative direct immunohistochemical (IHC) staining in improving the diagnosis accuracy in difficult cases of bronchiolar adenoma (BA). Methods: Nineteen cases with single or multiple pulmonary ground-glass nodules or solid nodules indicated by imaging in Cancer Hospital of Chinese Academy of Medical Sciences from January to July 2021 and with difficulty in differential diagnosis at frozen HE sections were selected. In the experimental group, direct IHC staining of cytokeratin 5/6 (CK5/6) and p63 was performed on frozen sections to assist the differentiation of BA from in situ/micro-invasive adenocarcinoma/adenocarcinoma/invasive mucinous adenocarcinoma. In the control group, two pathologists performed routine frozen HE section diagnosis on these 19 cases. The diagnostic results of paraffin sections were used as the gold standard. The sensitivity and specificity of BA diagnosis, consistency with paraffin diagnosis and time used for frozen diagnosis were compared between the experimental group and the control group. Results: The basal cells of BA were highlighted by CK5/6 and p63 staining. There were no basal cells in the in situ/microinvasive adenocarcinoma/adenocarcinoma/invasive mucinous adenocarcinoma. In the experimental group, the sensitivity and specificity with aid of direct IHC staining for BA were 100% and 86.7%, respectively, and the Kappa value of frozen and paraffin diagnosis was 0.732, and these were significantly higher than those in the control group (P<0.05). The average time consumption in the experimental group (32.4 min) was only 7 min longer than that in the control group (25.4 min). Conclusions: Direct IHC staining can improve the accuracy of BA diagnosis intraoperatively and reduce the risk of misdiagnosis, but require significantly longer time. Thus frozen direct IHC staining should be restricted to cases with difficulty in differentiating benign from malignant diseases, especially when the surgical modalities differ based on the frozen diagnosis.
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Humans , Paraffin , Sensitivity and Specificity , Adenocarcinoma in Situ , Adenoma/diagnosis , Adenocarcinoma, Mucinous/surgery , Frozen Sections/methodsABSTRACT
The pharmaceutical manufacturing model is gradually changing from intermittent manufacturing to continuous manufacturing and intelligent manufacturing. This paper briefly reviewed the supervision and research progress in continuous pharmaceutical manufacturing in China and abroad and described the definition and advantages of continuous pharmaceutical manufacturing. The continuous manufacturing of traditional Chinese medicine(TCM) at the current stage was summarized in the following three terms: the enhancement of the continuity of intermittent manufacturing operations, the integration of continuous equipment to improve physical continuity between units, and the application of advanced process control strategies to improve process continuity. To achieve continuous manufacturing of TCM, the corresponding key technologies, such as material property characterization, process modeling and simulation, process analysis technology, and system integration, were analyzed from the process and equipment, respectively. It was proposed that the continuous manufacturing equipment system should have the characteristics of high speed, high response, and high reliability, "three high(H~3)" for short. Considering the characteristics and current situation of TCM manufacturing, based on the two dimensions of product quality control and production efficiency, a maturity assessment model for continuous manufacturing of TCM, consisting of operation continuity, equipment continuity, process continuity, and quality control continuity, was proposed to provide references for the application of continuous manufacturing technology for TCM. The implementation of continuous manufacturing or the application of key continuous manufacturing technologies in TCM can help to systematically integrate advanced pharmaceutical technology elements and promote the uniformity of TCM quality and the improvement of production efficiency.
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Medicine, Chinese Traditional , Reproducibility of Results , China , Quality Control , Pharmaceutical PreparationsABSTRACT
Spermatogenic dysfunction caused by cyclophosphamide (CP) chemotherapy has seriously influenced the life quality of patients. Unfortunately, treatments for CP-induced testicular spermatogenic dysfunction are limited, and the molecular mechanisms are not fully understood. For the first time, here, we explored the effects of bone marrow mesenchymal stem cell-derived exosomes (BMSC-exos) on CP-induced testicular spermatogenic dysfunction in vitro and in vivo. BMSC-exos could be taken up by spermatogonia (GC1-spg cells). CP-injured GC1-spg cells and BMSC-exos were cocultured at various doses, and then, cell proliferation was measured using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay. In addition, photophosphorylation of extracellular-regulated kinase (ERK), p38 mitogen-activated protein kinase (p38MAPK), and protein kinase B (AKT) proteins was evaluated by western blotting as well as apoptosis in GC1-spg cells measured using flow cytometry. Treatment with BMSC-exos enhanced cell proliferation and reduced apoptosis of CP-injured GCI-spg cells. Phosphorylated levels of ERK, AKT, and p38MAPK proteins were reduced in CP-injured spermatogonia when co-treated with BMSC-exos, indicating that BMSC-exos acted against the reproductive toxicity of CP via the p38MAPK/ERK and AKT signaling pathways. In experiments in vivo, CP-treated rats received BMSC-exos by injection into the tail vein, and testis morphology was compared between treated and control groups. Histology showed that transfusion of BMSC-exos inhibited the pathological changes in CP-injured testes. Thus, BMSC-exos could counteract the reproductive toxicity of CP via the p38MAPK/ERK and AKT signaling pathways. The findings provide a potential treatment for CP-induced male spermatogenic dysfunction using BMSC-exos.
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Accurate methods for identifying pelvic lymph node metastasis (LNM) of prostate cancer (PCa) prior to surgery are still lacking. We aimed to investigate the predictive value of peripheral monocyte count (PMC) for LNM of PCa in this study. Two hundred and ninety-eight patients from three centers were divided into a training set (n = 125) and a validation set (n = 173). In the training set, the independent predictors of LNM were analyzed using univariate and multivariate logistic regression analyses, and the optimal cutoff value was calculated by the receiver operating characteristic (ROC) curve. The sensitivity and specificity of the optimal cutoff were authenticated in the validation cohort. Finally, a nomogram based on the PMC was constructed for predicting LNM. Multivariate analyses of the training cohort demonstrated that clinical T stage, preoperative Gleason score, and PMC were independent risk factors for LNM. The subsequent ROC analysis showed that the optimal cutoff value of PMC for diagnosing LNM was 0.405 × 109 l
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The effect of Danhong Injection on the endogenous metabolites of rabbit platelets was analyzed by the liquid chromatography-mass spectrometry( LC-MS) based metabonomic approach. Anti-platelet aggregation was detected after Danhong Injection treatment and the changes of platelet metabolites were analyzed by metabonomics. Principal component analysis( PCA) and partial least squares discriminant analysis( PLS-DA) were performed to investigate the effect of Danhong Injection on endogenous metabolites of platelets,characterize the biomarkers,and explore the relevant pathways and the underlying mechanism. As demonstrated by the pharmacodynamic results,Danhong Injection of different doses and concentrations antagonized platelet aggregation in a dose-and concentration-dependent manner. In contrast to the control group,25 differential metabolites such as nicotinic acid,nicotinic acid riboside,and hypoxanthine were screened out after platelets were treated by Danhong Injection. These metabolites,serving as important biomarkers,were mainly enriched in the nicotinic acid-niacinamide metabolic pathway and purine metabolic pathway. This study explored the therapeutic mechanism of Danhong Injection from a microscopic perspective by metabonomics,which is expected to provide a new idea for the investigation of platelet-related mechanisms.
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Animals , Rabbits , Biomarkers , Blood Platelets , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal/pharmacology , Metabolomics , TechnologyABSTRACT
The quality of traditional Chinese medicine tablets is correlated with clinical efficacy and drug safety, and plays a great role in promoting the development of traditional Chinese medicine. However, the existing traditional artificial identification and modern instrument detection in terms of accuracy and timeliness have both advantages and disadvantages. Therefore, how to quickly and accurately identify the quality of traditional Chinese medicine tablets has become a high-profile issue. The purpose of this paper is to explore the feasibility of the application of electronic eye technology in the study of rapid identification of traditional Chinese medicine quality. A total of 80 batches of samples were collected and tested by Fritillariae Cirrhosae Bulbus for traditional empirical identification(M_1) and modern pharmacopeia(M_2). The optical data was collected from electronic eyes, and the chemical metrology was used to establish suitable discrimination models(M_3). Four authenticity and commodity specification models, namely identification analysis(DA), minimum bidirectional support vector machine(LS-SVM), partial minimum two-multiplier analysis(PLS-DA), main component analysis identification analysis(PCA-DA), were established, respectively. The accuracies of the authenticity identification models were 82.5%, 90.0%, 96.2% and 93.8%, while the accuracies of the commodity specification identification models were 89.3%, 96.0%, 90.7% and 97.3%, respectively. The models were well judged, the authenticity identification was based on the final identification model of PLS-DA, and the commodity specification was based on the final identification model of PCA-DA. There was no significant difference between its accuracy and M_1, and the time of determination was much shorter than M_2(P<0.01). Therefore, electronic-eye technology could be used for the rapid identification of the quality of Fritillariae Cirrhosae Bulbus.
Subject(s)
Drugs, Chinese Herbal , Fritillaria , Medicine, Chinese Traditional , Plant Roots , TechnologyABSTRACT
Objective:To study the effects of various extracts of Tongkat Ali on uric acid excretion and renal function in rats with hyperuricemia. Methods:Totally 60 male Wistar rats were equally divided into the blank group,the model group,the positive control group,chloroform group,ethyl acetate group and n-butanol group. The rats were fed with yeast extract,adenine and potassium oxonate to make the model of hyperuricemia. After the animal model was made successfully,the other groups continued to give the modeling a-gent except for the blank group,and the other groups were given the corresponding medicine except for the model group after giving the modeling agent for one hour. After the model agent was given for 14 days,the levels of uric acid(UA),urea nitrogen(UN) and cre-atinine (Cr) in serum and urine and the activity of xanthine oxidase (XOD) in serum and liver were measured. Results: Compared with those in the model group,the levels of uric acid in chloroform group and n-butanol group were significantly lower than those in the model group (P<0.05),besides,the levels of uric acid and creatinine in urine in chloroform group and n-butanol group significantly increased (P<0.05). Compared with that in the model group,the XOD activity of chloroform group and n-butanol group significantly decreased (P<0.01). Compared with those in the positive control group,the levels of UA,Cr and UN in serum and urine and the ac-tivity of XOD in serum and liver in chloroform group, ethyl acetate group and n-butanol group had no significant differences (P >0.05). The levels of UA,Cr and UN in serum and urine and the activity of XOD in serum and liver in chloroform group,ethyl acetate group and n-butanol group had no significant differences (P>0.05). Conclusion: The chloroform extract and n-butanol extract in Tongkat Ali can reduce the serum level of uric acid in rats with hyperuricemia induced by adenine, yeast extract and potassium ox-onate,and the function may be related with the activity inhibition of xanthine oxidase.
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Objective To evaluate the clinical application of continuous two-layer suture of gallbladder incision with a single absorbable suture on laparoscopic minimally invasive gallbladder-preserving cholecystolithotomy. Methods The clinical data of 74 cases underwent laparoscopic minimally invasive gallbladder-preserving cholecystolithotomy were retrospectively analyzed. Main surgical procedures included the longitudinal incision of gallbladder wall, choledochoscopy and the removal of all stones and the closure of the gallbladder incision. The mucous incision was first closed using a 4-0 absorbable suture with continuous everting suture. Using the same suture, the seromuscular incision was then closed with continuous invering suture. The operation time, suturing time, complications and postoperative hospitalization time were also documented. Results Laparoscopic gallbladder-preserving cholecystolithotomy was successfully performed in all cases using the suturing technique introduced in Methods. The operation time was 33~78 min (average 45.11 ± 14.96 min). Suturing time for gallbladder incision was 9 ~ 22 min (average 14.86 ± 3.88 min). No severe complications occurred, such as bile leakage, peritonitis, residual gallstone, hemorrhage or infection. The postoperative hospitalization time was 2~4 d (average 3.21 ± 0.69 d). A postoperative follow-up of 3 ~ 62 months (average 35.50 ± 18.94 months) indicated gallbladder stone recurrence of 2 cases, with a recurrence rate of 2.7%. Continuous two-layer suture of gallbladder incision with a single absorbable suture is a safe, practical and reliable technique for the closure of the gallbladder incision in laparoscopic gallbladder-preserving cholecystolithotomy.
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Objective To analyze the curative effect of fossa intercondyloidea of micro-fracture and comminution of subcutaneous fat pad on meniscal tear healing under arthroscopy. Methods During the clinical operation, the samples of the micro-fracture and fat pad comminution were attracted to the outside of the joint. The source and properties of cells were identified based on preclinical medicine experiment in vitro. In clinical research, 75 cases of the simple meniscus injury were selected from January 2014 to January 2016 and divided into three groups refer to treatment: Group A, B and C. Different treatments were used in the three groups. Group A - micro-fracture group; Group B - fat pad group; Group C - blank group. All patients' conditions were assessed with Lysholm, VAS score and adverse event (Swelling, anterior knee pain). Results In the preclinical medicine experiment, cells released by micro fracture were fibroblast-like cells which tend to grow more rapidly in vortex shape. Cells released by comminution of fat pad contained a small amount of impurities and necrotic cells and are mostly similar to fibroblast in long spindle shape. Besides, the Human MSC Analysis Kit was used to authenticate two cells which were considered to be as BMSCs and IPFPSCs. In clinical research, all patients received an average of 10 (8 ～ 11) months follow-up visit. No severe complication, such as blood-vessel and nerve injury, infection occurred after operations in the patients followed. Before receiving the treatment, difference in Lysholm, VAS score and general condition of joint of Group A and B, C showed no statistical difference (P > 0.05). Compared with those before operation and after operation in both of four groups, the Lysholm and VAS score were improved (P < 0.05). Besides, there had significantly difference between group A and C, B and C after post treatment in Lysholm and VAS score (P < 0.05), however, group A and B did not. Compared group A and C, the early prevalence of adverse event of group B was higher, but last follow-up show no difference between three groups. Conclusion Combined with basic experimental and clinical studies, our research suggest that micro-fracture and comminution of fat pad have good effect on the repair of meniscus injuries. Nevertheless, it should be concerned that early prevalence of increased swelling amd anterior knee pain of comminution of fat pad.
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Objective:To investigate the effect of long non-coding BANCR on the migration and invasion of melanoma cells and its mechanism.Methods:qPCR was used to detect the expression of BANCR in melanoma patients and the relationship between clini-copathological data.Kaplan-Meier analysis the survival of melanoma patients with different expression of BANCR.Transwell invasion assay was used to detect the effect of BANCR on the invasive ability of melanoma cells.The effect of BANCR on the migration of melanoma cells was detected by scratch healing assay.Western blot was used to detect Wnt/β-catenin Signal pathway protein expression.Results:BANCR was highly expressed in melanoma,especially with the higher pathological stage or the lymph node metastasis.The higher expression of BANCR,the worse survival of melanoma patients.The inhibition of BANCR expression could reduce the invasion and migration ability of melanoma cells.The expression of β-catenin and c-myc protein in Wnt/β-catenin signaling pathway was down-regulated after silencing BANCR.Conclusion:Long non-coding BANCR was highly expressed in melanoma patients and was negatively correlated with survival time,it also regulates melanoma cell migration and invasion by activating the Wnt/β-catenin signaling pathway.
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AIM:To study the effect of Lycium barbarum polysaccharides (LBP) on oxidative stress injury of human endothelium-like EA. Hy926 cells induced by hydrogen peroxide (H2O2). METHODS:The EA. Hy926 cell model of oxidative stress injury was established by H2O2 treatment. The EA. Hy926 cells were divided into 5 groups:control group, damage (H2O2 at 50 mmol/L) group, LBP (100 mg/L) group, anti-damage groups (LBP at 50 mg/L, 100 mg/L or 200 mg/L+50 mol/L H2O2), and LY294002 (20 μmol/L) group. The effect of LBP at different concentrations on the cell viability of EA. Hy926 cells was measured by CCK-8 assay, and the optimum concentration of LBP was screened out. The apoptotic of EA. Hy926 cells was analyzed by flow cytometry. Acridine orange/ethidium bromide ( AO/EB) staining was used to observe the morphological characteristics of the apoptotic cells. The cell migration ability was detected by scratch method. The levels of nitric oxide (NO) and vascular endothelial growth factor (VEGF) in the cell culture medium were examined. The protein levels of cleaved caspase-3, Bax, Bcl-2, endothelial NO synthase (eNOS), p-eNOS and p-Akt were determined by Western blot. RESULTS:LBP at concentration of 100 mg/L significantly attenuated the injury of EA. Hy926 cells induced by H2O2, as indicated by improved cell viability ( P <0.05 ) and decreased apoptosis ( P <0.05). Pretreatment with LBP elevated the levels of NO and VEGF (P<0.05), and promoted the migration ability of EA. Hy926 cells. LBP also increased the Bcl-2/Bax ratio, down-regulated the protein level of cleaved caspase-3, and up-regulated the protein levels of eNOS and p-eNOS. The protective effect of LBP were abolished by pretreatment of the EA. Hy926 cells with the inhibitor of PI3K (P<0.05). As a result, the protein level of p-Akt was down-regulated, and the level of NO was also significantly reduced. CONCLUSION:LBP has protective effect on H2O2-induced EA. Hy926 cells by attenuating apoptosis of the cells. The mechanism is closely related to the activation of PI3K/Akt/eNOS signaling pathway.
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AIM: To study the effects of cladribine on growth and secretion activity of human umbilical vein endothelial cell line EA.hy926, and to investigate the mechanism of its anti-tumor effect by inhibiting endothelial cells. METHODS:The effects of cladribine at different concentrations on the cell viability were detected by CCK -8 assay.Apop-tosis and cell cycle distribution were examined by flow cytometry.The protein expression levels were determined by Western blot.The levels of tumor necrosis factor-α(TNF-α), transforming growth factor-β1(TGF-β1)and vascular endothelial growth factor(VEGF)secreted by EA.hy926 cells with cladribine treatment for 48 h were analyzed by ELISA.The nitric oxide(NO)production was measured by Gries method.RESULTS:Cladribine at 0.4~1 μmol/L inhibited the viability of EA.hy926 cells in time-and dose-dependent manners.The IC50was about 3.644 μmol/L.The results showed 43.74% cells in S phase when the concentration of cladribine was 0.4 μmol/L,and 77.23 % cells in S phase when the concentra-tion of cladribine was 1 μmol/L.The apoptosis was not induced by cladribine at 0.4~10 μmol/L.The protein expression of Bax and caspase-3 did not change.The expression of p21 increased and the p53 decreased(P<0.05).The levels of TNF-αand TGF-β1 secreted by EA.hy926 cells increased after cladribine treatment for 48 h.The levels of VEGF and NO decreased.CONCLUSION:Cladribine obviously inhibits the viability of EA.hy926 cells.The mechanism is related to the cell cycle arrest.Cladribine promotes the secretion of TNF-αand TGF-β1 by EA.hy926 cells and inhibits the secretion of VEGF and NO.
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AIM:To explore the regulatory effect of chemokine CCL 3 on exosome secretion from human bone marrow mesenchymal stem cells(hBMSCs).METHODS: hBMSCs were stimulated with chemokine CCL 3 at different concentrations in vitro.The proliferation of hBMSCs was measured by CCK-8 assay and viable cell counting.Exosome se-cretion from hBMSCs was qualitatively analyzed by transmission electron microscope(TEM)and flow cytometry, and the quantitative analysis was carried out by flow cytometry and nanoparticle tracking analysis(NTA).RESULTS:Compared with control group,the viability of the hBMSCs detected by CCK-8 assay was increased when hBMSCs were treated with CCL3(P<0.05).The results of viable cell counting demonstrated that the number of hBMSCs was raised in CCL 3 group in a dose-dependent manner(P<0.05).The results of flow cytometry showed that hBMSCs expressed 3 CCL3-related spe-cific receptors,CCR1,CCR5 and CCR9.Compared with control group,the fluorescence intensity of CCR9 in CCL3 group was obviously enhanced.However,no significant difference of fluorescence intensity for CCR 5 and CCR1 was observed be-tween the 2 groups.The results of NTA demonstrated that the secretion capacity of CCL 3-induced hBMSCs was far less than that in control group(P<0.05).However, the microvesicles larger than 100 nm in CCL3 groups were increased(P<0.05).The above results indicated that the higher concentration of CCL 3 induced the lower secretion of exosomes.In addi-tion,the results of flow cytometry demonstrated that CCL 3-induced hBMSCs showed lower quantity of CD 9 +exosomes than those in control group(P<0.01).CONCLUSION:CCL3 promotes the proliferation of hBMSCs but depresses the secre-tion of exosomes in a dose-dependent manner.CCL3 affects the size distribution of exosomes and increases the number of nonfunctional microvesicles of larger than 100 nm in size.CCL3 induces the expression of CCR9 in hBMSCs.
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Objective · To explore the association between cerebrospinal fluid (CSF) protein level and peripheral nerve demyelination in patients with Guillain-Barré syndrome (GBS). Methods · Clinical and biochemical data of 86 patients with GBS were retrospectively analyzed. According to electromyograms examination of peripheral nerve, GBS patients were divided into group with demyelination and group with axonal degeneration, and their clinical and biochemical characteristics were compared between the two groups. The correlation between CSF protein level and peripheral nerve demyelination was assessed by Spearman's correlation analysis. Results · Between the group with demyelination and group with axonal degeneration,there was no significant difference in gender, age, Hughes score, respiratory infection, gastrointestinal infection, erythra, ganglioside sodium injection and immunoglobulin G (IgG) index (P>0.05). Significant higher level of CSF protein, CSF albumin/serum albumin, IgG, and 24 h IgG intrathecal synthesis rate were detected in group with demyelination than that of in group with axonal degeneration (P<0.01). CSF protein level was positively correlated with peripheral nerve demyelination (r=0.345, P=0.001). Conclusion · The incidence of peripheral nerve demyelination increased accompanied with CSF protein level, and analysis of CSF protein level may be helpful in investigating the immunologic mechanism of peripheral nerve demyelination in GBS patients.
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Objective · To explore the association between cerebrospinal fluid (CSF) protein level and peripheral nerve demyelination in patients with Guillain-Barré syndrome (GBS). Methods · Clinical and biochemical data of 86 patients with GBS were retrospectively analyzed. According to electromyograms examination of peripheral nerve, GBS patients were divided into group with demyelination and group with axonal degeneration, and their clinical and biochemical characteristics were compared between the two groups. The correlation between CSF protein level and peripheral nerve demyelination was assessed by Spearman's correlation analysis. Results · Between the group with demyelination and group with axonal degeneration,there was no significant difference in gender, age, Hughes score, respiratory infection, gastrointestinal infection, erythra, ganglioside sodium injection and immunoglobulin G (IgG) index (P>0.05). Significant higher level of CSF protein, CSF albumin/serum albumin, IgG, and 24 h IgG intrathecal synthesis rate were detected in group with demyelination than that of in group with axonal degeneration (P<0.01). CSF protein level was positively correlated with peripheral nerve demyelination (r=0.345, P=0.001). Conclusion · The incidence of peripheral nerve demyelination increased accompanied with CSF protein level, and analysis of CSF protein level may be helpful in investigating the immunologic mechanism of peripheral nerve demyelination in GBS patients.
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AIM:To explore the effect of dasatinib on the viability, migration, cell cycle and apoptosis of human bone marrow mesenchymal stem cells (hBMSCs), as well as the underlying signal pathway to evaluate the influence of dasatinib on hematopoietic microenvironment clinically.METHODS:The cell viability was measured by CCK-8 assay.The migration ability was detected by wound healing assay.The cell cycle and apoptosis were analyzed by flow cytometry.Acridine orange/ethidium bromide staining was also used to detected apoptosis.The secretion of transforming growth factor-β1 (TGF-β1) and tumor necrosis factor-α (TNF-α) were measured by ELISA.The protein levels of cleaved caspase-3, protein kinase B (Akt) and phosphorylated Akt were determined by Western blot.RESULTS:Compared with control group, dasatinib at 1~10 nmol/L suppressed the viability and migration ability of hBMSCs, and dasatinib at concentration of 7 nmol/L was adopted in the following assays.Dasatinib promoted apoptosis, and blocked the cell cycle in G1 phase.In addition, the secretion of TGF-β1 and TNF-α was increased markedly.The protein levels of cleaved caspase-3 was increased, but the protein levels of Akt and phosphorylated Akt were decreased.CONCLUSION:Dasatinib inhibits the viability and migration ability of hBMSCs in a dose-dependent manner, promotes the secretion TGF-β1 and TNF-α, and induces cell cycle arrest and apoptosis.Dasatinib might regulate the biological behaviors of hBMSCs observed above by modulating the expression and phosphorylation of Akt.
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Blending process, which is an essential part of the pharmaceutical preparation, has a direct influence on the homogeneity and stability of solid dosage forms. With the official release of Guidance for Industry PAT, online process analysis techniques have been more and more reported in the applications in blending process, but the research on endpoint detection algorithm is still in the initial stage. By progressively increasing the window size of moving block standard deviation (MBSD), a novel endpoint detection algorithm was proposed to extend the plain MBSD from off-line scenario to online scenario and used to determine the endpoint in the blending process of Chinese medicine dispensing granules. By online learning of window size tuning, the status changes of the materials in blending process were reflected in the calculation of standard deviation in a real-time manner. The proposed method was separately tested in the blending processes of dextrin and three other extracts of traditional Chinese medicine. All of the results have shown that as compared with traditional MBSD method, the window size changes according to the proposed MBSD method (progressively increasing the window size) could more clearly reflect the status changes of the materials in blending process, so it is suitable for online application.
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The aim of this study was to develop a murine model of brain injury induced by high altitude hypoxic inflammation. In the study, we used a decompression chamber to mimic an acute hypobaric hypoxia, and 8-week-old male C57BL/6 mice were intraperitoneally injected with 5 mg/kg lipopolysaccharide (LPS) to induce inflammatory response. We determined the levels of pro-inflammatory factors (IL-6, TNF-α) and anti-inflammatory factor (IL-10) in mice serum using ELISA assays to confirm the high altitude hypoxic inflammation, and verified the brain injury after the inflammation using hematoxylin-eosin (HE) staining. The results showed that, among four experiment groups (ctrl, acute hypobaric hypoxia, LPS, and acute hypobaric hypoxia plus LPS groups), the acute hypobaric hypoxia plus LPS treatment group displayed the highest levels of IL-6, TNF-α, and IL-10. Meanwhile, the acute hypobaric hypoxia plus LPS treatment group showed the most severe cortex and hippocampus injuries, including cellular swelling, the widened pericellular spaces, angiogenesis, and shrunken neurons with darkly stained pyknotic nuclei, etc. Strikingly, nuclei ventrales posteriors thalami were found to be more sensitive to acute hypobaric hypoxia plus LPS treatment, and their destroy degrees were higher than those neurons in cortex and hippocampus. These results suggested that we established a reliable murine model of brain injury induced by high altitude hypoxic inflammation, and might be useful to the relevant studies.
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Animals , Male , Mice , Altitude , Brain Injuries , Cerebral Cortex , Disease Models, Animal , Hippocampus , Hypoxia , Inflammation , Mice, Inbred C57BL , NeuronsABSTRACT
<p><b>Objective</b>To explore the application value of real-time contrast-enhanced ultrasound (RTCEU) in improving the detection rate of transrectal ultrasound-guided prostate biopsy.</p><p><b>METHODS</b>This prospective study included 91 male patients with abnormally high PSA (4－20 μg/L) or abnormalities in DRE or MRI, who underwent 12+X prostate biopsy following conventional transrectal ultrasonography (TRUS) and RTCEU examination. We compared the numbers of suspected prostatic nodules before and after RTCEU as well as the detection rates of prostate cancer between conventional TRUS-guided 12PBx and 12PBx plus lesion-targeted biopsy procedures.</p><p><b>RESULTS</b>Totally, 57 of the 86 suspected lesions on TRUS (66.3%), and 108 of the 118 abnormal nodules on RTCEU (91.5%) were confirmed to be prostate cancer. RTCEU achieved a significantly higher detection rate than TRUS (P<0.01). A total of 39 cases of prostate cancer (42.8%) were detected by RTCEU, while only 28 (30.7%) by TRUS, with statistically significant difference in the detection rate between the two procedures (P=0.033).</p><p><b>CONCLUSIONS</b>Real-time contrast-enhanced ultrasound can significantly improve the detection rate of prostate cancer and provide a valuable guide to targeted prostate biopsy.</p>