Dynamic change and significance of Mindin protein in chronic hepatitis B treated with PEG-IFNα-2b / 临床肝胆病杂志

ObjectiveTo investigate the change and potential role of Mindin protein in the treatment of chronic hepatitis B （CHB） with PEG-IFNα-2b. MethodsA total of 29 CHB patients who received the treatment with PEG-IFNα-2b in The Second Affiliated Hospital of Xi’an Jiaotong University from January 2018 to December 2019 were enrolled， and according to their clinical outcome， they were divided into cured group with 17 patients and uncured group with 12 patients. Peripheral blood samples were collected from both groups at baseline， 12 weeks， and 24 weeks to measure blood routine indices， liver function parameters， hepatitis B markers， and Mindin protein. HBsAg， alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， and Mindin protein at different time points were compared between the two groups. The independent-samples t test was used for comparison of normally distributed continuous data between two groups， and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups； a Spearman correlation analysis was used to investigate correlation； a multiple linear regression analysis was used to investigate the influence of HBsAg and ALT on the content of Mindin protein. ResultsThe analysis of baseline data showed that there were significant differences in the levels of HBsAg， HBeAb， albumin， and albumin/globulin ratio between the cured group and the uncured group （all P<0.05）. The cured group tended to have a gradual increase in the level of Mindin， and the level of Mindin at 24 weeks was significantly higher than that at baseline （P<0.05）. The cured group had a significantly higher level of Mindin protein than the uncured group at 24 weeks （P=0.019）. The cured group had a significantly lower level of HBsAg than the uncured group （P<0.05）， with a significant change from baseline to each time point within the cured group （P<0.05）. In addition， the levels of ALT and AST in the cured group tended to first increase and then decrease， and the expression levels at 12 weeks were significantly higher than those at baseline （P<0.05）. At 12 weeks， there was a strong linear correlation between Mindin protein levels and ALT in the untreated group （r=0.760 8， P<0.05）， and further multiple linear regression analysis also demonstrated a linear relationship between the two （b=1.571， P=0.019）. ConclusionThere is a significant difference in the level of Mindin protein between the cured group and the non-cured group after 24 weeks of PEG-IFNα-2b antiviral treatment， and therefore， detecting the dynamic changes of Mindin protein can better predict the treatment outcome of CHB， which provides a reference for clinical practice.

Expression level and clinical significance of SOCS1 in the serum of patients with chronic HBV-related liver disease / 西安交通大学学报(医学版)

【Objective】 To study the expression levels of suppressor of cytokine signaling 1 (SOCS1) and its clinical significance in hepatitis B virus (HBV)-related liver diseases. 【Methods】 For this study we enrolled 25 patients with chronic hepatitis B (CHB), hepatitis B cirrhosis, or HBV-associated chronic acute liver failure (HBV-ACLF), and 25 healthy controls. The expression levels of SOCS1 mRNA in peripheral blood mononuclear cells (PBMCs) were determined using the RT-PCR method. The levels of SOCS1 and interleukin-6 (IL-6) in the plasma of patients with chronic liver diseases and healthy controls were measured using the ELISA method. The relative expression levels of SOCS1, SOCS1 mRNA, and other laboratory test indicators such as HBV-DNA, alanine aminotransferase (ALT), aspartate aminotransferase (AST), prothrombin activity (PTA) and total bilirubin (TBil) were compared among the groups. Additionally, the correlation between the expression levels of SOCS1 mRNA and the aforementioned laboratory indicators was assessed. 【Results】 The expression levels of SOCS1 mRNA and serum SOCS1 were highest in the HBV-ACLF group, followed by the cirrhosis group, and lowest in the healthy control group, with statistically significant differences (F=109.65, P<0.001). The relative expression of SOCS1 mRNA was positively correlated with TBil (r=0.89, P<0.001), ALT (r=0.89, P<0.001), AST (r=0.84, P<0.001) and IL-6 (r=0.93, P<0.001), but negatively correlated with PTA (r=-0.89, P<0.001) and was not significantly correlated with HBV-DNA (P=0.28). 【Conclusion】 The expression levels of SOCS1 in patients with HBV-related chronic liver diseases can reflect the severity of the disease and show a significant correlation with indicators used to assess the severity of liver diseases.

Application of magnetic compression anastomosis to colorectal anastomosis and reconstruction / 中华消化内镜杂志

Clinical cases treated by magnetic compression anastomosis (MCA) for different causes and types of intestinal stenosis/ atresia to successfully achieve intestinal recanalization were reviewed, so as to explore the clinical application of MCA. From May 2019 to August 2022, 4 patients underwent colorectal MCA for intestinal recanalization in the First Affiliated Hospital of Xi'an Jiaotong University and Northwest Women and Children's Hospital. All operations went well, and the intestinal anastomosis was recanalized. The magnetic ring was discharged in 7-15 days, and the postoperative colonoscopy or radiography showed that the anastomosis was intact. MCA can be used to treat different types of colorectal stenosis and atresia due to different reasons, and can also be used to assist intestinal anastomosis in colorectal surgery.

A tactical nanomissile mobilizing antitumor immunity enables neoadjuvant chemo-immunotherapy to minimize postsurgical tumor metastasis and recurrence

Neoadjuvant chemotherapy has become an indispensable weapon against high-risk resectable cancers, which benefits from tumor downstaging. However, the utility of chemotherapeutics alone as a neoadjuvant agent is incapable of generating durable therapeutic benefits to prevent postsurgical tumor metastasis and recurrence. Herein, a tactical nanomissile (TALE), equipped with a guidance system (PD-L1 monoclonal antibody), ammunition (mitoxantrone, Mit), and projectile bodies (tertiary amines modified azobenzene derivatives), is designed as a neoadjuvant chemo-immunotherapy setting, which aims at targeting tumor cells, and fast-releasing Mit owing to the intracellular azoreductase, thereby inducing immunogenic tumor cells death, and forming an in situ tumor vaccine containing damage-associated molecular patterns and multiple tumor antigen epitopes to mobilize the immune system. The formed in situ tumor vaccine can recruit and activate antigen-presenting cells, and ultimately increase the infiltration of CD8+ T cells while reversing the immunosuppression microenvironment. Moreover, this approach provokes a robust systemic immune response and immunological memory, as evidenced by preventing 83.3% of mice from postsurgical metastasis or recurrence in the B16-F10 tumor mouse model. Collectively, our results highlight the potential of TALE as a neoadjuvant chemo-immunotherapy paradigm that can not only debulk tumors but generate a long-term immunosurveillance to maximize the durable benefits of neoadjuvant chemotherapy.

Experimental study of magnetic compression technique for anastomosis reconstruction of esophagus / 中国胸心血管外科临床杂志

@#Objective    To investigate the feasibility of magnamosis rings designed based on magnetic compression technique in esophageal anastomosis reconstruction. Methods    According to the anatomical characteristics of esophagus in SD rats, the esophageal magnamosis rings were designed. SD rats were used as animal models (n=10, 5 males and 5 females) to complete the magnetic anastomosis reconstruction of the cervical esophagus using magnamosis rings, and the operation time, animal survival, postoperative complications, magnetic rings excretion time were recorded. Two weeks after operation, the rats were killed, and the esophageal anastomotic specimens were obtained. The blasting pressure of the anastomotic site was measured and the formation of the anastomotic site was observed with naked eyes. Results    Esophageal magnamosis was successfully performed in 10 SD rats, and the median operation time was 11 (8-13) min. All rats survived without anastomotic leakage, anastomotic stenosis, or magnetic rings incarceration. The magnetic rings were discharged after 8 (5-10) days and the burst pressure was higher than 300 mm Hg. Visual observation showed that the anastomotic muscle healed well and the mucosa was smooth. Conclusion    The magnetic compression technique  can be used for anastomosis reconstruction of esophagus, which has the advantages of simple operation and reliable anastomosis effect, and has clinical application prospect.

ToxR represses the synthesis of c-di-GMP in Vibrio parahaemolyticus / 生物工程学报

Vibrio parahaemolyticus, the main pathogen causing seafood related food poisoning worldwide, has strong biofilm formation ability. ToxR is a membrane binding regulatory protein, which has regulatory effect on biofilm formation of V. parahaemolyticus, but the specific mechanism has not been reported. c-di-GMP is an important second messenger in bacteria and is involved in regulating a variety of bacterial behaviors including biofilm formation. In this study, we investigated the regulation of ToxR on c-di-GMP metabolism in V. parahaemolyticus. Intracellular c-di-GMP in the wild type (WT) and toxR mutant (ΔtoxR) strains were extracted by ultrasonication, and the concentrations of c-di-GMP were then determined by enzyme linked immunosorbent assay (ELISA). Three c-di-GMP metabolism-related genes scrA, scrG and vpa0198 were selected as the target genes. Quantitative real-time PCR (q-PCR) was employed to calculate the transcriptional variation of each target gene between WT and ΔtoxR strains. The regulatory DNA region of each target gene was cloned into the pHR309 plasmid harboring a promoterless lacZ gene. The recombinant plasmid was subsequently transferred into WT and ΔtoxR strains to detect the β-galactosidase activity in the cellular extracts. The recombinant lacZ plasmid containing each of the target gene was also transferred into E. coli 100λpir strain harboring the pBAD33 plasmid or the recombinant pBAD33-toxR to test whether ToxR could regulate the expression of the target gene in a heterologous host. The regulatory DNA region of each target gene was amplified by PCR, and the over-expressed His-ToxR was purified. The electrophoretic mobility shift assay (EMSA) was applied to verify whether His-ToxR directly bound to the target promoter region. ELISA results showed that the intracellular c-di-GMP level significantly enhanced in ΔtoxR strain relative to that in WT strain, suggesting that ToxR inhibited the production of c-di-GMP in V. parahaemolyticus. qPCR results showed that the mRNA levels of scrA, scrG and vpa0198 significantly increased in ΔtoxR strain relative to those in WT strain, suggesting that ToxR repressed the transcription of scrA, scrG and vpa0198. lacZ fusion assay showed that ToxR was able to repress the promoter activities of scrA, scrG and vpa0198 in both V. parahaemolyticus and E. coli 100λpir. EMSA results showed that His-ToxR was able to bind to the regulatory DNA regions of scrA and scrG, but not to the regulatory DNA region of vpa0198. In conclusion, ToxR inhibited the production of c-di-GMP in V. parahaemolyticus via directly regulating the transcription of enzyme genes associated with c-di-GMP metabolism, which would be beneficial for V. parahaemolyticus to precisely control bacterial behaviors including biofilm formation.

Design of Magnamosis System for Endoscopic Tissue Clamping / 中国医疗器械杂志

Based on the principle of magnetic anastomosis technique, the design of magnetic anastomosis system for endoscopic tissue clamping is proposed. The system includes a semi-ring magnet, a special structure transparent cap and a detachable push rod. With the help of the existing digestive endoscopy and endoscopic tissue gripper, the endoscopic close clamping and anastomosis of the bleeding or perforated tissue can be completed. After the anastomosis, the magnet falls off and is discharged through the digestive tract. Animal experiments showed that the system was easy to use, the fistula was clamped firmly, the magnet was discharged for 7~21 days, and there was no magnet retention and digestive tract obstruction. Further safety verification, optimization of endoscopic operation, the system can be used in clinical trial.

Transcriptional regulation of pilABCD by OpaR in Vibrio parahaemolyticus / 中华微生物学和免疫学杂志

Objective:To study the transcriptional regulation of pilABCD by the master quorum sensing (QS) regulator OpaR in Vibrio parahaemolyticus. Methods:Total RNAs were extracted from the wild type (WT) and opaR mutant (Δ opaR) strain. Quantitative real-time PCR (qPCR) was employed to calculate the transcriptional variation of pilA (the first gene of pilABCD operon) between WT and Δ opaR. The regulatory DNA region of pilABCD was cloned into the corresponding restriction endonuclease sites of pHRP309 harboring a promoterless lacZ reporter gene. The recombinant pHRP309 plasmid was then transferred into WT and Δ opaR, respectively, to detect the β-galactosidase activity in cellular extracts using a β-Galactosidase Enzyme Assay System (Promega). The primer extension assay was applied to map the transcription start site of pilABCD using the total RNAs extracted from the WT strain as the template. The regulatory DNA region of pilABCD was amplified by PCR, and the over-expressed His-OpaR was purified under native conditions with nickel loaded HiTrap Chelating Sepharose columns (Amersham). Thereafter, the electrophoretic mobility shift assay (EMSA) was applied to analyze the DNA-binding activity of His-OpaR to the target DNA in vitro, and the DNase I footprinting assay was further employed to detect the DNA-binding sites of His-OpaR within the target DNA. Results:The results of qPCR and LacZ fusion assays showed that OpaR activated the transcription of pilABCD, leading to a gradual increase in the expression level of pilA with the extension of culture time. The primer extension assay detected only one transcription start site located at 155 bp upstream of pilA. The results of EMSA and DNase Ⅰ footprinting assays showed that His-OpaR protected two DNA regions located from -246 to -197 bp and -181 to -131 bp upstream of pilA. Conclusions:Vibrio parahaemolyticus OpaR activated the transcription of pilABCD in a direct manner.

Research progress of SEPT9 gene in tumor / 国际检验医学杂志

Septin 9 (SEPT9) gene, a member of the conserved framework protein genes family, has guanosine triphosphataseg (GTPase) activity and play an important role in a wide range of biological processes such as cell division, cell polarization and membrane remodeling.Accumulated evidence have confirmed that SEPT9is closely related to the generation and development of human diseases.And SEPT9has become the new aroused general interest in tumor related research at this stage.This article mainly reviews the structural features of SEPT9gene, action mechanism of SEPT9protein and the role of SEPT9gene in a variety of common oncogenesis, and explores the potential value of SEPT9as a marker in early tumor screening.

intravoxel incoherent motion MRI for prediction of histological grade and muscle invasion in bladder urothelial carcinoma / 中国医学影像技术

Objective To observe the utility value of MR intravoxel incoherent motion (IVIM) in histological grading and muscle invasion of bladder urothelial carcinoma.Methods According to postoperative histologic grade and T staging,60 patients with bladder urothelial carcinoma confirmed by surgery and pathology were divided into low grade (LG) group and high grade (HG) group,as well as muscle-noninvasive bladder cancer (NMIBC) or muscle-invasive bladder cancer (MIBC) group.MR IVIM parameters (apparent diffusion coefficient standard [ADCst],true diffusion coefficient [D],pseudodiffusion coefficient [D*] and perfusion fraction [f]) were compared with independent-samples t tests.A binary Logistic regression model was established to evaluate the predicted probability of combined IVIM parameters.ROC curves of IVIM parameters and their combination's predicted probability were drawn,and the diagnostic efficiency was evaluated.Results ADCst,D and f values of HG group were significantly lower than those of LG group (all P＜0.05).Area under ROC curve (AUCs) for ADCst,D and f value to differentiate HG from LG were 0.88,0.86 and 0.72,respectively (all P＜0.01),and AUCs for predicted probability of combined ADCst and D,combined ADCst and f and combined D and f were 0.91,0.90 and 0.88,respectively (all P＜0.0001).ADCst,D and f values of M1BC group were significantly lower than those of NMIBC group (all P＜0.0001).AUCs for ADCst,D and f value to differentiate MIBC from NMIBC were 0.91,0.85 and 0.88,respectively (all P＜0.0001),and all AUCs for predicted probability of combined ADCst and D,combined ADCst and f and combined D and f were both 0.93 (all P＜0.000 1).Conclusion Lower ADCst,D and f values may indicate greater possibility of high grade and muscle invasion of bladder urothelial carcinoma.Combination of IVIM parameters can improve diagnostic efficacy.

Research progress of febrile seizures / 中国医师进修杂志

Febrile seizures (FS) is the most common convulsive disorder in childhood and is a frequent problem for pediatricians.Most of the children have a good prognosis, but some children have recurrent attacks. Severe cases can even develop into epilepsy, which can cause great damage to the physical and mental health of children.Further understanding and studying issues related to febrile seizures has become a major concern for pediatricians and many families.This article reviews the definition, incidence, classification, etiology, pathogenesis, and epilepsy correlation and management measures of febrile seizures, and is expected to be applied in clinical practice to provide guidance for the prevention, diagnosis and treatment of febrile seizures.

Evaluation of the practical efficacy of a colloidal gold detection reagent of rabies virus antibody / 中华实验和临床病毒学杂志

Objective@#To evaluate the practical efficacy of a colloidal gold (CG) detection reagent of rabies virus antibody.@*Methods@#Series dilutions of rabies immunoglobulin and serum samples of rabies vaccine immunized population were tested by a CG detection reagent of rabies virus antibody and rapid fluorescent focus inhibition test (RFFIT). The consistency of the qualitative results of rabies virus antibodies between the two methods were compared. The comparison of rates was made by Chi-square test.@*Results@#For rabies immunoglobulin diluent, the detection limit of the rabies virus antibody CG detection reagent was higher than 6.53 IU/ml but lower than 9.53 IU/ml. For the serum samples, the detection limit of the rabies virus antibody CG detection reagent was higher than 2.80 IU/ml but lower than 3.30 IU/ml. The positive rates of serum rabies virus antibodies detected by CG and RFFIT were 26% and 67% respectively, and the difference was statistically significant (χ2 =13.66, P=0.000). The results of RFFIT were regarded as gold standard, false negative results but no false positive results were observed when CG was used to detect serum rabies virus antibodies.@*Conclusions@#The sensitivity of the rabies virus antibody CG detection reagent is poor, and attention should be paid to the phenomenon of missing some practically positive results in practical application of CG detection reagent.

Clinical correlation between incipient C-reactive protein and kidney injury of anti-neutrophil cytoplasmic antibody associated vasculitis / 中华肾脏病杂志

Objective To investigate the relationship between the incipient serum C-reactive protein (CRP) and clinicopathologic features in anti-neutrophil cytoplasmic antibody associated vasculitis (AAV).Methods Data of 138 consecutive AAV patients were collected.According to their serum CRP levels,patients were divided into group 1 with normal CRP,group 2 with slightly increased CRP and group 3 with severely increased CRP.Clinical features of AAV and histopathologic features of the kidney injury were compared among groups.Results CRP levels increased in 77.53％ AAV patients on admission.Patients in the group of severely increased CRP had the highest levels of BVAS,serum C3,serum ANCA titer,leukocyte counts and the lowest levels of hemoglobin and albumin among the 3 groups (all P ＜ 0.05).The mortality during the stage of therapy was highest in patients with severely increased CRP (P ＜ 0.05).The focal kidney damage was more obvious in patients with severely increased CRP.There was no significant difference in renal prognosis among patients with different CRP levels.Conclusion The levels of incipient serum C-reactive protein of AAV vary in different patients and are positively correlated with patients' inflammation status as well as the disease activity,but are not correlated with the severity of kidney injury.

The effect of triptolide on HIF1αand VEGF expressions under hypoxia in human umbilical vein endothelial cells / 天津医药

Objective To investigate the effect of triptolide (TP) on the expression of hypoxia inducible factor 1 alpha (HIF1α) and vascular endothelial growth factor (VEGF) in the human umbilical vein endothelial cells (HUVECs) under hypoxia. Methods (1) HUVECs were treated with 0, 40, 80, 160 and 320 nmol/L TP (named with hypoxia group, TP40 group, TP80 group, TP160 group and TP320 group, respectively) under the hypoxic condition (37℃, 5%CO2, 1%O2, 94%N2) for culturing 12 hours. Meanwhile, cells cultured under normoxia condition (without TP added) were set as the normoxia group. Western blot assay was used to detect the expression of HIF1αin each group. (2) The cells were divided into normal control group, hypoxia group and TP80 group. The immunofluorescence method was performed to detect the localization of HIF1α in cells. (3) Expressions of VEGF were detected by Western blot assay in TP80 group and hypoxia group. (4) The cells were divided into hypoxia group, TP80 group, TP80+KF20 group (80 nmol/L TP and 20μmol/L KC7F2), and TP80+KF30 group (80 nmol/L TP and 30 μmol/L KC7F2). After 12-hour culturing, Western blot assay was used to detect the expressions of HIF1α and VEGF in each group. Results (1) Under the normoxia condition, no HIF1α was detected in HUVECs. The expression level of HIF1αwas significantly increased in TP80 group than that in hypoxia group (P<0.05), while there was no significant change in expression of hypoxia HIF1αin TP160 group and TP320 group compared with that of hypoxic group. (2) The immunofluorescence result showed that HIF1α was mainly expressed in the nucleus. (3) The expression of VEGF was significantly increased in TP80 group than that in hypoxia group (P < 0.05). (4) After the intervention of KC7F2, HIF1αand VEGF expression levels were significantly decreased in the TP80+KF20 group and the TP80+KF30 group than those in the TP80 group (P<0.05). Conclusion TP can improve the expression of HIF1αand VEGF to accelerate the proliferation of endothelial cells under hypoxia condition.

Clinical Efficacy of Compound Honeysuckle Decoction Hot and Humid Joint Fusidic Acid Cream in Treatment of Targeted Drugs-induced Rash / 现代生物医学进展

Objective:To analyze the clinical efficacy of compound honeysuckle decoction hot and humid joint fusidic acid cream in the treatment of targeted drugs-induced rash.Methods:80 cases of patients with targeted drugs-induced rash admitted in our hospitalfrom August 2014 to August 2016 were selected and divided into two groups with 40 cases in each group according to the drawing method.The control group was treated by fusidic acid cream,while the observation group was treated by compound honeysuckle decoction hot and humid joint fusidic acid cream,the changes of symptom score,quality of life after treatment,clinical efficacy and incidence of adverse reactions were compared between two groups.Results:After treatment,the symptoms score of observation group ((6.87± 1.25) points) was lower than that of the control group ((10.29± 2.74) points)(P＜0.05),the quality of life score ((3.15± 0.57)points) of observation group was lower than that of the control group ((6.42± 1.20) points)(P＜0.05).The effective rate of observation group(95.00％) was higher than that of the control group(77.50％)(P＜0.05),no statistical difference was found in the incidence of adverse reactions was observed between two groups (P＞0.05).Conclusion:Compound honeysuckle decoction hot and humid joint fusidic acid cream was effective in the treatment of targeted drugs-induced rash,it could effectively relieve the clinical symptoms,improve the quality of life.

Rescuing Bombyx mori bidensovirus in BmN cells in vitro / 生物工程学报

Bombyx mori bidensovirus (BmBDV) has been identified as causing chronic densonucleosis in Bombyx mori specifically. The replication mechanism of BmBDV remains unknown. Its genome comprises two single stands DNA (VD1 and VD2). In order to rescue infectious virions in vitro, we obtained the total viral DNA extracted from the BmBDV-infected larvae midguts, subsequently cloned the full-length sequence of BmBDV genome fragments by PCR and constructed recombinant plasmids pMD18T-VD1 and pUC-VD2. The linear genome fragments were obtained by digesting recombinant plasmids with corresponding restriction enzymes, and then collectively transfected BmN cells by the method of liposome-embedding. We determined the replication of the virus gene by PCR with the template of demethylated total DNA extracted from the post-transfect BmN cells. Meanwhile, we collected the total proteins from the post-transfect BmN cells and the larvae midgut of feeding the post-transfect BmN cells to perform Western blotting analysis, and detected the expression of viral genes. Here we firstly confirm that infectious virions can be rescued in BmN cells by linear co-transfect method.

Regulating Effects of Emodin on the Expression of Aquaporin-3 in Tongue Tissues of Rats with Experimental Severe Acute Pancreatitis / 中国中医药信息杂志

Objective To investigate the expression of aquaporin-3 (AQP3) in tongue tissues of rats with experimental severe acute pancreatitis (SAP) and regulating effects of emodin. Methods Rat models with SAP were established by injecting 5% sodium taurocholate retrogradely into gallbladder and pancreas. SD male rats were randomly divided into sham-operation group, model group, and emodin group. After model establishment, rats in the emodin group received gavage with emodin 20 mg/kg each day, while rats in the model group and sham-operation group received gavage with normal saline. The mental state, thick greasy tongue fur and mortality of rats were observed every day after model establishment, and 5 days later, protein and genetic expression of AQP3 were detected by Western blot and RT-PCR. Results Compared with the model group, the mortality and the thick greasy tongue fur significantly decreased, the protein and genetic expression of AQP3 significantly decreased in the emodin group (P<0.05). On the 5th day, 11 rats in the model group survived, and 5 rats had thick greasy tongue fur. Compared with the sham-operation group, the protein and genetic expression of AQP3 in the model group were higher (P<0.05). Conclusion Emodin can improve the severity of SAP and decrease the incidence of thick greasy tongue fur significantly by reducing the protein and genetic expression of AQP3.

Discriminant Analysis of Platform Parameters of Rhesus at Different Stages of SIV/SAIDS / 广州中医药大学学报

Objective To predict the disease progression risks of healthy rhesus ( normal) and rhesus infected with simian immunodeficiency virus ( SIV) in the stages of long-term nonprogressor ( LTNP) , normal progressor ( NP) , rapid progressor ( RP) by discriminant analysis. Methods Five-year observation was carried out in SIV infected rhesus model without any intervention. The SIV infected rhesus model at the stages of LTNP, NP, RP were selected, 10 in each group, and T lymphocyte subsets and serum parameters for spleen-deficiency syndrome and kidney-deficiency syndrome in SIV infected rhesus were compared with 5 healthy monkey having the same survival time. The influence factors of different types of disease progression were screened from T cell subsets and Chinese medical syndrome indexes, and then the discriminant equation was established to predict the risks. Results White blood cell ( WBC) count and lymphocyte ( LYM) ratio were enrolled into the discriminant equation before infection, and T4 level and Log10RNA of set point were enrolled into the discriminant equation in the platform period. The test results for the uniform rate of the established discriminant function showed that the total coincidence rate of theoretic distinguish to the actual data was 57.1% , 91.2%respectively before infection and in the platform period. Conclusion The pre-infection WBC count and LYM ratio can be used as a reference for the evaluation of different types of disease progresson, and Log10RNA and T4 level at platform phase can be used as the predicting factors of different types of disease progression risk prediction.

Comparison of different ways to trace BMSCs after cerebral ischemia-reperfusion injury / 天津医药

Objective To compare different ways to trace bone marrow mesenchymal stem cells (BMSCs) after being transplanted in cerebral ischemia-reperfusion injury. Methods Male SD rats of SPF grade were randomly divided into sham group, model group (ischemia-reperfusion,IR), BrdU tracing group, PKH26 tracing group and GFP tracing group. Fo?cal cerebral ischemia-reperfusion model was established by blocking middle cerebral artery. 24 hours after cerebral isch?emia-reperfusion injury, 10μL BMSCs that were labeled respectively by BrdU, PKH26, GFP were added respectively into BrdU, PKH26 and GFP tracing group while equal volum of normal saline was added into sham group and model group. Mod?el and transplanting cells efficacy was determined by neural behavioral score, TTC staining and brain water content;Neurons were counted using tar violet staining;The number of transplant cells in the transplanting site was assessed by fluorescence microscopy. Results Before transplanting, there was no significant difference among BrdU, PKH26 and GFP group in cell labeled efficacy. By contrast, neural behavioral score, brain infarct volume and brain tissue water content were significantly lower in all three tracing groups than that in model group 4 weeks after transplantation while neuron counts were markedly higher. There was no significant difference of above parameters among the three tracing groups. However, the number of traced transplanting cells in damaging area in GFP group is significantly higher than that in BrdU group and PKH26 group. Conclusion In cerebral ischemia-reperfusion injury, the tracing effect of GFP last longer, therefore it is significantly more effective than BrdU and PKH26.

Effects of ZHX3 gene silence on the expression of osteoblast-related factors in BMSCs / 天津医药

Objective To investigate the effects of zinc fingers and homeoboxes 3 (ZHX3) silence on expressions of smad3, smad4 and RUNX2 in bone marrow mesenchymal stem cells (BMSCs). Methods ZHX3 low expression vector (ZHX3 silent group) was constructed and was transfected to rat BMSCs. Empty vector was transfected into BMSCs and was used as vehicle control group, and wild type BMSCs was used as the control group. The cell transfection rate was measured under a fluorescence microscope, and then the successful transfection was identified. The immunocytochemistry and immu?noblotting methods were used to detect the expression levels of smad3, smad4 and RUNX2. Results (1) Cells with BMSCs phenotype can be obtained by recovery culturing. (2) After transfection, the green fluorescent protein was found in ZHX3 si?lence group and vehicle control group. Blank control group showed no significant fluorescence. The expression level of ZHX 3 was significantly lower in ZHX3 silence group than that of vehicle control group. (3) Results of immunofluorescence asssay showed that the positive expressions of smad3 and smad4 were located in nucleus and cytoplasm, the positive expression of RUNX2 was mainly located in nucleus. Positive cells were observed in three groups. There was no significant difference in fluorescence intensity between the control group and the vehicle control group, but the fluorescence intensity was significant?ly lower in ZHX3 gene silence group than that of two control groups. (4) There were no significant differences in expressions of smad3, smad4 and RUNX2 betweem control group and the vehicle control group, but they were significantly higher than those of ZHX3 silence group(P < 0.05). Conclusion ZHX3 gene silence can delay vitro osteogenesis of BMSCs, which may play a role by the down-regulated expression levels of smad3, smad4 and RUNX2.