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OBJECTIVES@#To study the clinical application effect of "kindergarten effect" in radiotherapy for children with tumor based on the psychology of preschool children aged 3-5 years.@*METHODS@#A total of 30 children, aged 3-5 years, who were admitted to the Department of Radiotherapy, Tianjin Medical University Cancer Institute and Hospital, from January 2020 to August 2021 were enrolled in this prospective study. The children were randomly divided into a control group and a test group, with 15 children in each group. The children in the test group were treated in "kindergarten mode", i.e., all children were treated together at a specified time and left together after all children completed treatment. Those in the control group were treated alternately with adult patients according to the treatment time based on the type of radiotherapy fixation device. The treatment compliance was evaluated for both groups, and the two groups were compared in terms of the setup errors in the superior-inferior (SI), left-right (LR), and anterior-posterior (AP) directions.@*RESULTS@#Compared with the control group, the test group showed a significantly shorter time for finishing the treatment (P<0.05) and a significantly lower proportion of children with treatment interruption (P<0.05). Compared with the control group, the test group showed smaller mean errors in the SI, LR and AP directions after image-guided radiotherapy, with significant differences in the mean errors in the SI and LR directions (P<0.05).@*CONCLUSIONS@#With the application of the "kindergarten effect", most children can actively cooperate in radiotherapy, and it can also improve the accuracy and repeatability of positioning and help to achieve the desired treatment outcome.
Subject(s)
Adult , Humans , Neoplasms/radiotherapy , Prospective Studies , Radiotherapy Planning, Computer-AssistedABSTRACT
Type 2 diabetes mellitus (T2DM) is a metabolic disease with an increasing incidence worldwide, which leads to damage to various tissues and organs including the liver. MiR-31 is conserved across species and closely associated with metabolic diseases, but its role in type 2 diabetic liver injury has not been elucidated. This study aimed to investigate the effect of miR-31 on liver injury in type 2 diabetes and its underlying mechanism. Four to six weeks old male FVB mice and miR-31-positive transgenic mice were randomly divided into FVB mice control group (C), FVB mice induced diabetes group (DM) and miR-31-overexpression transgenic mice induced diabetes group (31DM). After 1 week of adaptive feeding, the T2DM mouse model was induced by high-fat feeding combined with intraperitoneal injection of streptozotocin (STZ) for 6 weeks. The general condition of mice and related metabolic indicators showed that the increased food and water intake, weight loss and glucose and lipid metabolism disorders could be reversed by miR-31 in T2DM mice. HE staining and liver histological activity index (HAI) scoring results showed that miR-31 improved the inflammatory status in the liver tissue of T2DM mice and decreased the HAI score. RT-qPCR results showed that the high expression of miR-31 was accompanied by a decrease in the expression of activating transcription factor 6 (ATF6) mRNA in the liver of T2DM mice. Furthermore, Western blotting results showed that miR-31 inhibited the expression of endoplasmic reticulum stress-related proteins such as ATF6, glucoregulatory protein 78 (GRP78) and C/EBP homologous protein (CHOP) in the liver of T2DM mice. In conclusion, miR-31 may ameliorate liver injury in T2DM mice by regulating glucose and lipid metabolism disorders and insulin resistance, and inhibiting endoplasmic reticulum stress factors such as ATF6, GRP78, and CHOP.
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BACKGROUND@#Shenque (CV8) acupoint is located on the navel and has been therapeutically used for more than 2000 years in Traditional Chinese Medicine (TCM). However, clinical research on the underlying therapeutic molecular mechanisms of the CV8 acupoint lags far behind. This study aimed to study the mechanisms of umbilical acupoint therapy by using stem cells.@*METHODS@#The morphological characteristics of CV8 acupoint were detected under a stereomicroscope using hematoxylin and eosin (H&E) staining. Oil Red, Masson, and immunohistochemical staining on multi-layered slices were used to identify the type of cells at the CV8 acupoint. Cell proliferation was measured by a cell counting kit-8 (CCK-8) method. Flow cytometry and immunohistochemistry were used for cell identification. Induced differentiation was used to compare the differentiation of cells derived from CV8 acupoint and non-acupoint somatic stem cells into other cell types, such as osteogenic, adipogenic, and neural stem cell-like cells.@*RESULTS@#Morphological observations showed that adipose tissues at the linea alba of the CV8 acupoint in mice had a mass-like distribution. Immunohistochemical staining confirmed the distribution of stem cell antigen-1 (Sca-1) positive cells in the multi-layered slices of CV8 acupoint tissues. Cells isolated from adipose tissues at the CV8 acupoint exhibited high expression of Sca-1 and CD44 and low expression of CD31 and CD34, and these cells possessed osteogenic, adipogenic, and neurogenic stem cell-like cell differentiation ability. The cell proliferation (day 4: 0.5138 ± 0.0111 vs. 0.4107 ± 0.0180, t = 8.447, P = 0.0011; day 5: 0.6890 ± 0.0070 vs. 0.5520 ± 0.0118, t = 17.310, P 100 μm: 2.6000 ± 0.5477 vs. 0.8000 ± 0.8367, t = 4.025, P = 0.0038) were significantly enhanced in somatic stem cells derived from the CV8 acupoint compared to somatic stem cells from the groin non-acupoint. However, cells possessed significantly weaker osteogenicity ([2.697 ± 0.627]% vs. [7.254 ± 0.958]%, t = 6.893, P = 0.0023) in the CV8 acupoint group.@*CONCLUSIONS@#Our study showed that CV8 acupoint was rich with adipose tissues that contained abundant somatic stem cells. The biological examination of somatic stem cells derived from the CV8 acupoint provided novel insights for future research on the mechanisms of umbilical therapy.
Subject(s)
Animals , Mice , Acupuncture Points , Adipose Tissue , Adult Stem Cells , Cell Differentiation , Cells, Cultured , OsteogenesisABSTRACT
Neural tube defects (NTDs) are congenital defect diseases caused by cell proliferation and apoptosis disorders. Using RNA-Seq assays, we found the increased expression of DNA damage-inducible transcript 4 (Ddit4) in embryonic brain tissues from NTD fetuses. In this study, we intend to explore the effects of Ddit4 overexpression on the proliferation and apoptosis of HT-22 cells and related mechanisms to lay the foundation for the study of the role of Ddit4 in NTDs. According to the mouse Ddit4 sequence, we constructed the eukaryotic expression vector pEX-3-Ddit4. The results of restriction enzyme analysis and sequencing showed that the eukaryotic expression vector pEX-3-Ddit4 was successfully constructed. qRT-PCR and Western blotting results showed that the expression level of Ddit4 in HT-22 cells was significantly increased after transfection of PEX-3-Ddit4 (P < 0. 01) . CCK-8 and Western blotting results showed that Ddit4 overexpression decreased the proliferation of HT-22 cells (P < 0. 01) . Flow cytometry showed that Ddit4 overexpression increased the proportion of cells in the G
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Lysine acetylation has emerged as one of the most important post-translational modifications that participates in various biological and pathological processes. Histone acetyltransferase 1 (HAT1) as the first identified protein ε-amino lysine acetyltransferase is able to regulate the acetylation of histones and non-histone proteins. However‚ the acetylation substrates and sites mediated by HAT1 in liver cancer are poorly understood. In this study‚ we demonstrated that HAT1 was highly expressed in the liver cancer tissues‚ which was negatively associated with the prognosis of patients. Based on the establishment of the HAT1-knockout HepG2 cell line‚ we employed a quantitative proteomics approach to study the profiling of acetylation mediated by HAT1 in HepG2 cells. Interestingly‚ we identified a total of 858 Kac sites on 547 proteins in the HepG2 cell line‚ in which HAT1 mediated the levels of Kac of 74 sites on 68 proteins. The pathways and metabolic processes that were affected by HAT1-dependent acetylation modification were analyzed by bioinformatics. The results show that Kac regulates disease development‚ RNA biology‚ spliceosome and nucleosome assembly‚ oxidative stress‚ various signaling pathways and metabolic pathways‚ etc.. Moreover‚ we verified that the HAT1-mediated acetylation modification could promote abnormal lipid metabolism. CCK8 assays‚ clone formation and Edu assays revealed that HAT1 could remarkably enhance the cell proliferation of liver cancer in vitro. Thus‚ our finding explored the profiling of HAT1-mediated protein acetylation in HepG2 cells‚ which provides new insights into the underlying mechanism by which HAT1 mediates the development of liver cancer. Clinically‚ the HAT1-mediated acetylation sites could be used for the precise targets of drug development.
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Colorectal cancer is a common malignant tumor in the gastrointestinal tract, with the characteristics of high morbidity and mortality. Studies have shown that the occurrence and development of colorectal cancer is closely related to the abnormal activation of Wnt signaling pathway. Abnormal expression of β-catenin in Wnt pathway is found both in the cytoplasm and nucleus of tumor cells. Different drugs can target the Wnt signaling pathway and its upstream and downstream related factors to inhibit or suppress the development of colorectal cancer. We review the components of Wnt signaling pathway, and the correlation between Wnt signaling pathway and colorectal cancer. Then, we summarize the current status of drug research targeting the Wnt signaling pathway in colorectal cancer. Finally, the challenges and prospects of these methods and drugs were briefly summarized.
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OBJECTIVES@#To evaluate the effects of antimicrobial peptide GH12 designed @*METHODS@#The cariogenic three-species biofilm consis-ted of the cariogenic @*RESULTS@#The biomass and density of the cariogenic three-species biofilm treated with GH12 decreased compared with those of the control. The number of @*CONCLUSIONS@#GH12 can reduce the number of
Subject(s)
Humans , Biofilms , Dental Caries , In Situ Hybridization, Fluorescence , Pore Forming Cytotoxic Proteins , Streptococcus mutansABSTRACT
OBJECTIVE@#To observe the effect of acupoint stimulation on the quality of recovery in patients with radical thyroidectomy under the concept of enhanced recovery after surgery (ERAS).@*METHODS@#A total of 62 patients with radical thyroidectomy were randomized into an observation group and a control group, 31 cases in each one. In both of the two groups, general anesthesia with tracheal intubation was applied, the same anesthesia induction and maintenance medication were given. In the observation group, auricular point pressing with magnetic beads was adopted at bilateral shenmen (TF) and transcutaneous electrical acupoint stimulation (dilatational wave, 2 Hz/100 Hz in frequency, 6 to 12 mA) was performed at bilateral Hegu (LI 4) and Neiguan (PC 6) from 30 min before anesthesia induction to the end of the anesthesia. In the control group, medical adhesive plaster was pasted at bilateral shenmen (TF) and the electrodes were plastered at bilateral Hegu (LI 4) and Neiguan (PC 6) with no corresponding stimulation. In both of the two groups, visual analogue scale for anxiety (VAS-A) score was observed to evaluate the anxiety severity before anesthesia induction; the total intraoperative dosages of sufentanil, remifentanil and propofol were recorded; the numerical rating scale (NRS) score was used to assess the pain severity of instant time (T0) and 30 min (T1) of entering post-anesthesia recovery room (PACU), motor and static mode at 2 h (T2), 6 h (T3), 12 h (T4), 24 h (T5) after surgery; time of first anal exhaust, time of getting out of bed after surgery, total hospitalization time and the incidences of postoperative nausea and vomiting were observed; the quality of recovery was assessed by the 40-item quality of recovery score (QoR-40).@*RESULTS@#The VAS-A score and the total intraoperative dosage of remifentanil in the observation group were reduced compared with the control group (0.05). The time of first anal exhaust and getting out of bed after surgery in the observation group were advanced than those in the control group (0.05). Compared with the control group, the QoR-40 score was increased in the observation group (<0.05).@*CONCLUSION@#Acupoint stimulation can improve the preoperative anxiety in patients with radical thyroidectomy, reduce the intraoperative anesthetic dosage and postoperative pain, advance the time of anal exhaust and getting out of bed, improve the quality of postoperative recovery and enhance the recovery process.
Subject(s)
Humans , Acupuncture Points , Enhanced Recovery After Surgery , Postoperative Nausea and Vomiting , Thyroidectomy , Transcutaneous Electric Nerve StimulationABSTRACT
<p><b>Background</b>Measuring total serum calcium is important for the diagnosis of diseases. Currently, results from commercial kits for calcium measurement are variable. Generally, the performance of serum calcium measurements is monitored by external quality assessment (EQA) or proficiency testing schemes. However, the commutability of the EQA samples and calibrators is often unknown, which limits the effectiveness of EQA schemes. The aim of this study was to evaluate the bias of serum calcium measurements and the commutability of processed materials.</p><p><b>Methods</b>Inductively coupled plasma mass spectrometry was applied as a comparative method, and 14 routine methods were chosen as test methods. Forty-eight serum samples from individual patients and 25 processed materials were quantified. A scatter plot was generated from patient samples, and 95% prediction intervals were calculated to evaluate the commutability of the processed materials and measurement bias at three concentration levels was used to determine the accuracy of routine assays.</p><p><b>Results</b>All assays showed high precision (total coefficient of variation [CV] <2.26%) and correlation coefficients (r > 0.99). For all assays, the mean bias for the 48 patient samples ranged from -0.13 mmol/L to 0.00 mmol/L (-5.61-0.01%), and the ranges for the three concentrations were -0.10-0.04 mmol/L (-5.71-2.35%), -0.14--0.01 mmol/L (-5.80--0.30%), and -0.19-0.04 mmol/L (-6.24-1.22%). The EQA samples, calibrators, and animal sera exhibited matrix effects in some assays; human serum pools were commutable in all assays; certificate reference materials were commutable in most assays, and only GBW09152 exhibited a matrix effect in one assay; and aqueous reference materials exhibited matrix effects in most assays.</p><p><b>Conclusions</b>Biases for most assays were within the acceptable range, although the accuracy of some assays needs improvement. Human serum pools prepared from patient samples were commutable, and the other tested materials exhibited a matrix effect.</p>
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We developed a new method for chiral separation of fourteen amino alcohols by nonaqueous capillary electrophoresis (NACE) with the D-(+)-gluconic acid δ-lactone-boric acid complex as chiral selector. In order to achieve good enantioseparation, the effects of D-(+)-gluconic acid δ-lactone and boric acid concentrations, triethylamine concentration, as well as capillary temperature were systematically investigated. The optimized conditions were identified as follows:an uncoated fused silica capillary of 50 μm ID with a total length (Ltot) of 55 cm and an effective length (Leff) of 45 cm; 200 mmol·L-1 D-(+)-gluconic acid δ-lactone, 80 mmol·L-1 boric acid, and 57.4 mmol·L-1 triethylamine in methanol; positive pressure injection at 2.9 psi for 2 s; capillary temperature, 25 ±0.2℃; applied voltage, +15 kV; detection wavelength, 214 nm. Under the optimized conditions, a good chiral resolution was achieved in most of the tested drugs. This method provides a foundation for the development and application of new chiral selectors of polyhydroxy compound-boric acid complexes in chiral drugs analysis by NACE.
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<p><b>OBJECTIVE</b>The interleukin-23 receptor (IL-23R) has been shown to be associated with autoimmune diseases in many different populations. This study aimed to investigate the association between IL-23R gene polymorphism and susceptibility to Hashimoto's thyroiditis (HT) in Chinese Han population of Shandong.</p><p><b>METHODS</b>A case-control cohort study was performed in 145 HT patients from First People's Hospital of Jining between February 2010 to October 2013 and 150 healthy controls. Two single nucleotide polymorphisms located in the promoter region of IL-23R gene (rs17375018 and rs7517847) were examined by polymerase chain reaction-restriction fragment length polymorphism analysis. Hardy-Weinberg equilibrium was performed using the Chi-square test. Genotype frequencies were estimated by direct counting, and allele and genotype frequencies between patients and controls were analyzed by the Chi-square test.</p><p><b>RESULTS</b>The rs17375018 GG genotype and the G allele were significantly increased in HT patients compared with healthy controls (P = 0.034 and P = 0.013, respectively). No association was identified between HT patients and healthy controls in rs7517847.</p><p><b>CONCLUSION</b>The study demonstrated that polymorphism of IL-23R gene rs17375018 is highly associated with HT in Chinese Han population of Shandong, suggesting that IL-23R gene polymorphism (rs17375018 G) may play a critical role in susceptibility to HT.</p>
Subject(s)
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Asian People , Case-Control Studies , Cohort Studies , Genetic Predisposition to Disease , Genetics , Genotype , Hashimoto Disease , Genetics , Polymorphism, Single Nucleotide , Genetics , Receptors, Interleukin , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To explore the clinical effects of cervical posterior pedicle screw internal fixation and interbody fusion for the treatment of atlanto-axial vertebral instability fractures.</p><p><b>METHODS</b>From July 2008 to July 2013, 21 patients with atlanto-axial vertebral instability fractures were treated with vertebral pedicle screw internal fixation and interbody fusion through posterior approach. There were 14 males and 7 females, aged from 20 to 55 years old with an average of 32 years. Lifted and guided the atlanto-axial vertebral pedicle screw to reduce atlanto-axial vertebral displacement.</p><p><b>RESULTS</b>All patients were followed up from 6 to 24 months with an average of 12.5 months. Wounds got healed without complication of infection. Clinical symptoms were relieved at 6 months after operation. According to ASIA score standard to assess at 6 months after operation, the items of motion, light touch and needle score had obviously improved, and respectively were 99.45 ±0.27, 111.09 ± 0.47,111.11 ± 0.58. VAS and NDI scores also had obviously improved, and respectively were 1.04 ± 0.38 and 12.56 ± 2.24. Imaging examinations showed internal fixation locations were good, without atlanto-axial joint instability.</p><p><b>CONCLUSION</b>Cervical posterior pedicle screw internal fixation and interbody fusion can effectively restore the stability of atlanto-axial joint, reduce complication and obtain satisfactory effects.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Young Adult , Atlanto-Axial Joint , Wounds and Injuries , General Surgery , Fracture Fixation, Internal , Methods , Joint Instability , General Surgery , Pedicle Screws , Spinal Fractures , General Surgery , Spinal Fusion , MethodsABSTRACT
Background Adeno-associated virus-based vector is one of most efficient vehicles.It presents with a long-term and efficient transfer and expression of therapeutic genes with minimal toxicity.But its delayed-and low-efficient transgene expression limits the application of AAV vector.To explore an improving method of AAV infecting RPE cells is the hot spot. Objeetive Present study was to investigate whether adeno-associated virus (AAV)combined with low dose non-replieable adenovirus(Ad-null)can enhance its infection efficieney on RPE ceils in vitro. Methods Human RPE cells were isolated from the donate eyeballs under the approval of the Ethic Committee of this hospital.The cells were cultured in DMEM containing 10%fetal bovine serum.AAV particles with enhanced green fluorescence protein(EGFP)were added into the medium alone or in combination with different amount of adenovirus for 30 days.The cells were detected under the fluorescence microscope,and the protein expression levels of report gene EGFP in RPE cells were analyzed with Western blotting assay. Results Melanin granules could be found in cultured RPE cells.EGFP was expressed in RPE cells at 2 days after AAV-EGFP infection and peaked at 12 days and remained for about 3-week duration,showing the green influorescence under the influorescence mwroscope.After the cells were infected by AAV2-EGFP with 0.01 to 1000 MOI Ad-null respectively,the number of cells with green influorescence was obviously increased with the enhanced infiuorescence intensity.The enhance of the infection efficiency began in the 0.1 MOI Ad-null group and peaked in 10 MOI Ad-null group.Dead cells were exhibited in the 100 or more MOI Ad-nulor group.Western blotting assay demonstrated that the protein expression level of EGFP in RPE cells enhanced significantly in 1 and 10 MOI Ad-null groups compared with only AAV infection group. Conclusion These finding suggested that the infection efficiency of AAV can be improved significantly when it is used with low dose Ad-null in vitro.This offers a basis for further study of gene therapy.
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Objective To construct a new conditionally replicative adenovirus (CRAds) targeting carcinoembryonic antigen (CEA) positive colorectal cancer cells. Methods The DNA fragment of the CEA gene promoter was amplified through PCR and cloned into the vector carrying fusion reporter gene EGFP-Luc to construct expression plasmid pCEA-EGFPLuc. The constructed plasmid pCEA-EGFPLuc was transfected into CEA positive and negative cells by liposome. The activity of CEA gene promoter was evaluated by detecting the expression of EGFP and luciferase activity. The conditionally replicative adenovirus Ad.CEA-E1A/CMV-TK carrying suicide gene HSVtk was constructed, in which the E1A gene was controlled under CEA promoter. CEA positive(Lovo and SW620)and negative tumor cells(HeLa) were infected with Ad.CEA-E1A/ CMV-TK. The selective cytotoxicity of Ad.CEA-E1A/CMV-TK and the synergistic effect of the virus with GCV in CEA positive tumor cells were evaluated by the expression of E1A, cytopathic effect and cell survival rate. Results CEA promoter possesses a good specificity as well as high activity. The expression of E1A only presented in CEA positive tumor cells. After infection with Ad. CEA-E1A/CMV-TK, the cell survival rates of Lovo and SW620 were (36.72±2.49)% and (39.82±4.76)%, respectively, significantly lower than that of Hela[(87.44±2.76)%1 (P<0.01). When combined with GCV, Ad.CEA-E1A/CMV-TK had better oncolytic effect on Lovo and SW620 cells, with cell survival rates of (17.26±3.65) % and (23.93±5.40) %, respectively, significantly lower than those without GCV[(36.72±2.49) % and (39.82±4.76) %, respectively] (P<0.01). Conclusion Ad. CEA-E1A/CMV-TK under the control of CEA promoter has selective cytotoxic effect on CEA positive colorectal cancer cells, and the effect can be enhanced when combined with GCV.
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<p><b>OBJECTIVE</b>To provide anatomical basis for the internal fixation of scaphoid fractures.</p><p><b>METHODS</b>The shape and vascular lake of 48 dry scaphoids and 36 wet scaphoids were observed.</p><p><b>RESULTS</b>The data of dry bone group and wet bone group were as follows: the height of scaphoid tubercle were (11.28+/-0.94) mm and (10.35+/-1.54) mm; the thickness of scapoid waist were (12.02+/-1.90) mm and (11.21+/-1.20) mm; the width of scapoid waist were (10.59+/-1.11) mm and (11.34+/-1.47) mm; the minimal thickness of the body of scapoid were (6.51+/-1.22) mm and (8.54+/-1.07) mm; the axis length of scapoid were (25.68+/-2.21) mm and (26.50+/-2.56) mm; the width of epicondyle of scaphoid of distal portion, waist and proximal portion were (6.50+/-1.06) mm, (5.14+/-1.01) mm, (4.42+/-1.16) mm and (6.64+/-1.18) mm, (6.01+/-0.75) mm and (5.71+/-0.78) mm, respestively. The main blood vessels came from the dorsal and the radial of wrist and passed through the whole scaphoid along the crest of scaphoid.</p><p><b>CONCLUSION</b>The internal fixation of scaphoid can be designed according to the anatomical data without damaging the articular surface and blood supply.</p>
Subject(s)
Adult , Female , Humans , Male , Fracture Fixation, Internal , Methods , Wrist , Wrist Injuries , General SurgeryABSTRACT
<p><b>OBJECTIVE</b>To observe the immunity preventive action of Tripterygium wilfordii ployglycosidium (TWP) on type 1 diabetes mellitus (T1DM) in non-obese diabetic (NOD)mice and explore its possible mechanism.</p><p><b>METHODS</b>Changes of blood glucose, the incidence rate of T1DM and pancreatitis were observed and the expressions of IFN-gamma,TNF-alpha, IL-10mRNA in pancreas of mice detected by RT-PCR after NOD mice were given TWP by adopting cyclophosphamide accelerated induction of T1DM.</p><p><b>RESULTS</b>The mean blood glucose level in TWP group (10.73 mmol/L) was lower than that in the control group (20. 53 mmol/L) at the end of the experiment(P < 0.01 ). The incidence rate of T1DM in TWP group (43. 33% ) was lower than that in the control group (71.43% , P <0. 01). The mean grade of inflammation of pancreatic islet in TWP group (1.45 +/- 1. 11) was lower than that in the control group (2. 27 +/- 1.22, P <0. 05 ). In TWP group the number of insulin-positive cells was 242. 80 +/-168.93 ,which was more than that of control group with significant difference (95.60 +/- 39.55,P <0.05). The expressions of TNF-alpha mRNA and IFN--gamma mRNA were lower (P <0.01). No obvious change was found in IL-10 mRNA expression.</p><p><b>CONCLUSION</b>TWP could prevent the occurrence of T1DM in NOD mice, whose mechanism might be correlated to the down-regulating of the expression of Thl cytokines in pancreatic tissues.</p>
Subject(s)
Animals , Mice , Blood Glucose , Diabetes Mellitus, Type 1 , Drugs, Chinese Herbal , Therapeutic Uses , Hypoglycemic Agents , Therapeutic Uses , Interferon-gamma , Metabolism , Interleukin-10 , Metabolism , Mice, Inbred NOD , Pancreas , Metabolism , Protective Agents , Therapeutic Uses , Tripterygium , Tumor Necrosis Factor-alpha , MetabolismABSTRACT
Construction of mutant strain is an essential method in pathogenesis researches. The conventional method for Brucella unmarked deletion mutant construction is based on suicide plasmid, but the efficiency is very low. In the present study, we first optimized the electroporation parameters, and then, the cloning plasmid pEX18Gm containing sacB was successfully used to construct unmarked deletion mutant of the type IV secretion system. This indicated that by using conventional cloning plasmid as suicide plasmid in Brucella, unmarked deletion mutants can be constructed with high efficiency.
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Brucella organisms are facultative intracellular bacteria capable of surviving inside professional and non-professional phagocytes.Upon cell contact the bacteria is internalized via receptor molecules.Once inside cells,Brucella localizes in early phagosomes,where it avoids fusion with late endosomes and lysosomes.Then,the bacterium redirects its trafficking to autophagosomes and finally reaches the endoplasmic reticulum,the replicating niche.Once inside the endoplasmic reticulum,Brucella extensively replicates without restricting basic cellular functions or inducing damage to cells.Invasion,intracellular trafficking and replication of Brucella organisms in professional and non-professional phagocytes and the molecular determinants involving Brucella intracellular life are reviewed in this article.
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NOD mice were treated intraperitoneally with tripterygium wilfordii ployglycosidium (TWP) for 4 weeks to observe the incidence of cyclophosphamide accelerated diabetes.The apoptosis of?-cells was detected by TUNEL,the expression of caspase-3 in islets of the NOD mice was detected by immunohistochemistry and the expression of caspase-8 mRNA in pancreas by RT-PCR.The results revealed that the incidence of diabetes in TWP group was lower than that in control group.The apoptosis index of?-cells was decreased in TWP group. The expression of caspase-3 in islets and the expression of caspase-8 mRNA in pancreas in TWP group were lower than those in control group.