Application of Intestinal Organoids in The Study of Intestinal Disease Mechanism / 生物化学与生物物理进展

Intestinal organoids are constructed by crypts or stem cells from the intestine under the 3D support of the culture matrix. They contain all mature cells of the intestine, and have become a new and efficient platform for studying the mechanism of intestinal diseases. Compared with 2D cell culture, organoids can not only more effectively simulate the physiological structure and function of the intestine, but also better restore the true ecology of the intestine in different external environments. Therefore, it is more widely used in the study of pathogenesis of different intestinal diseases. This article reviewed the new progress of intestinal organoids culture, and the application and progress of intestinal organoids in the pathogenesis of inflammatory bowel diseases, colorectal cancer and celiac disease in recent years, and also discussed the application of intestinal organoids in drug research and development and screening.

Saikosaponin A Changes Drug Resistance of A549/DDP Cells Through Wnt/β-Catenin Pathway / 中国实验方剂学杂志

Objective:To investigate the effect and mechanism of saikosaponin A （SSA） on the reversal of cisplatin （DDP） resistance in human lung cancer cell line A549/DDP. Methods:The resistance of A549 and A549/DDP cells to DDP and the inhibitory effects of SSA against the proliferation of A549 and A549/DDP cells were detected using cell counting kit-8 （CCK-8）. The apoptosis rates of A549/DDP cells treated with SSA or DDP or SSA combined with DDP and the changes in reactive oxygen species （ROS） were determined by flow cytometry. The mRNA expression levels of C-myc， B-cell lymphoma 2 （Bcl-2） and cysteinyl aspartate-specific protease-3 （Caspase-3） were detected by real-time polymerase chain reaction （Real-time PCR）， followed by the determination of <italic>β</italic>-catenin transcriptional activity using the TopFish dual-luciferase reporter assay system and the measurement of <italic>β</italic>-catenin protein expression in A549/DDP cells by Western blot. Results:The results of CCK-8 assay showed that the DDP resistance of A549/DDP cells was 12.82 times that of A549 cells （<italic>P</italic><0.05）. SSA inhibited the viability of A549 cells with the half maximal inhibitory concentration （IC₅₀） being 34.9 μmol·L^-1， and also suppressed the viability of A549/DDP cells in a concentration-dependent manner. Since the inhibition rate of 20 μmol/L SSA against A549/DDP cells was less than 10%， the reversal concentration was set at 20 μmol/L. Flow cytometry revealed that compared with the control， DDP alone increased the apoptosis rate of A549/DDP cells （<italic>P</italic><0.05）， stimulated the accumulation of intracellular ROS （<italic>P</italic><0.05）， down-regulated the mRNA expression levels of C-myc and Bcl-2 in A549/DDP cells， up-regulated Caspase-3 mRNA expression， and reduced the transcriptional activity of <italic>β</italic>-catenin （<italic>P</italic><0.05）. Compared with the DDP group， the SSA+DDP group exhibited obviously increased apoptosis of A549/DDP cells， enhanced accumulation of intracellular ROS， down-regulated C-myc and Bcl-2 mRNA expression， up-regulated Caspase-3 mRNA expression （<italic>P</italic><0.05）， and weakened <italic>β</italic>-catenin transcription （<italic>P</italic><0.05）. DDP combined with SSA better decreased the <italic>β</italic>-catenin protein expression in contrast to that of control or DDP （<italic>P</italic><0.05）. Conclusions:SSA enhances the sensitivity of A549/DDP cells to DDP possibly by inhibiting the activation of Wnt/<italic>β</italic>-catenin pathway.

Duplex effect of CCK8 and PGE2 in CVB-attacked human peripheral blood pDC / 中国免疫学杂志

Objective:To investigate the immunomodulation of CCK8 on the Coxsackievirus B ( CVB )-attacked human peripheral blood plasmacytoid dendritic cells(pDC). Methods:Peripheral blood mononuclear cells of healthy volunteers were separated by Ficoll-Hypaque gradient density centrifugation. The pDC was separated and divided into five groups,which were the control group, CVB attacked group,the group of CCK8 treated after CVB attack,the group of PGE2 treated after CVB attack and the group of CCK8+PGE2 treated after CVB attack. 100-time TCID50 of CVB was applied for the attack on pDC. Real-time PCR and Immunofluorescence technique were employed to detect the expression of CCK1R/CCK2R mRNA and protein. Then,the expression levels of costimulatory molecules such as CD80,CD86,HLA-DR ligand,and the chemokine receptor CCR7 were evaluated by Flow Cytometry Analysis. The supernatants of pDCs were collected, and the content of IFN-α was determined by Enzyme-linked Immunosorbent Assay. Results:CCK1R and CCK2R were co-expressed in human peripheral blood pDC,and both were significantly upregulated after CVB attack in vitro. Expression of CD80,CD86,HLA-DR and IFN-α were decreased in the CVB+CCK8 group compared with the CVB group,which suggested that CCK8 may reduce the CVB activation of pDC. Whereas expression of CD80,CD86,HLA-DR,CCR7 and IFN-α were increased in the CVB+PGE2 group compared with the CVB group,which suggested that PGE2 may increase the CVB activation of pDC in vitro. Conclusion:CCK8 repressed the CVB-attacked pDC,while PGE2 activated the CVB-attacked pDC.

Effect of warm joint needling plus rehabilitation on the balance function and quality of life of patients with spastic hemiplegia after ischemic cerebral stroke / 针灸推拿医学（英文版）

Objective:To observe the effect of warm joint needling plus rehabilitation techniques on the balance function and quality of life (QOL) of patients with spastic hemiplegia after ischemic cerebral stroke.Methods:Ninety patients with spastic hemiplegia after ischemic cerebral stroke were randomized into a rehabilitation group,a warm joint needling group and an observation group,with 30 cases in each group.The rehabilitation group was intervened by Bobath therapy,the warm joint needling group was treated with joint needling on the affected side plus warm needling,and the observation group was given the same rehabilitation treatment as the rehabilitation group together with the same warm joint needling as the warm joint needling group.The three groups were treated once another day,1 month as a treatment course for 6 months.Before the treatment,and respectively after 2-week,1-month,3-month,and 6-month treatment,the modified Ashworth scale (MAS) was used to measure the anti-spasm ability of the lower limb,the Berg balance scale (BBS) was adopted to evaluate the balance function,and the stroke-specific quality of life scale (SS-QOL)was employed to estimate the QOL.Results:After 3-month and 6-month treatment,the lower-limb MAS scores in the observation group were significantly better than those in the rehabilitation group and the warm joint needling group (all P＜0.05).After 1-month,3-month and 6-month treatment,the BBS scores in the observation group were significantly better than those in the rehabilitation group and the warm joint needling group (all P＜0.05).After 2-week,1-month,3-month and 6-month treatment,the SS-QOL scores in the observation group were markedly better than those in the rehabilitation group and the warm joint needling group (all P＜0.05).Conclusion:Warm joint needling plus rehabilitation can effectively improve the lower-limb spasticity state,balance function and QOL in patients with spastic hemiplegia after ischemic cerebral stroke.

Clinical Observation of Relaxing Needling at Meridian-muscle Nodes plus PNF Method in Improving Upper-limb Motor Function and Quality of Life in Hemiplegic Shoulder Pain / 上海针灸杂志

Objective To observe the effect of relaxing needling at meridian-muscle nodes plus rehabilitation techniques on upper-limb motor function and quality of life (QOL) in hemiplegic shoulder pain after cerebral stroke. Method Ninety patients with hemiplegic shoulder pain after cerebral stroke were randomized into a rehabilitation group of 30 cases [intervened by proprioceptive neuromuscular facilitation (PNF) method], a relaxing needling group of 30 cases (intervened by relaxing needling at meridian-muscle nodes), and a comprehensive group of 30 cases (intervened by relaxing needling at meridian-muscle nodes plus PNF method), to receive 6-week treatment in total. Before and after the intervention, Assessment Face Scale (AFS), Fugl-Meyer Assessment Scale (FMA) and Stroke Specific Quality of Life Scale (SS-QOL) were evaluated, and the occurrence rate of shoulder-hand syndrome was assessed. Result After 1-week treatment, the AFS score of the comprehensive group was significantly different fromthat in the rehabilitation group and relaxing needling group (P<0.05). After 6-week treatment, the FMA and SS-QOL scores in the comprehensive group were significantly different from those in both rehabilitation group and relaxing needling group (P<0.05). Twelve weeks later, the occurrence rate of shoulder-hand syndrome in the comprehensive group was significantly different from that in the other two groups (P<0.05). Conclusion Relaxing needling at meridian-muscle nodes plus PNF method can effectively improve the efficacy in the treatment of hemiplegic shoulder pain, reduce the occurrence of shoulder-hand syndrome, and enhance the QOL, thus is an effective approach.

Program optimization in the stage treatment of periarthritis of shoulder with acupuncture, moxibustion and positive functional exercise / 中国针灸

<p><b>OBJECTIVE</b>To optimize the therapeutic programs for periarthritis of shoulder treated with acupuncture, moxibustion and kinetohterapy with orthogonal design method adopted.</p><p><b>METHODS</b>The orthogonal design table of L8 (2(7)) hierarchical principle was used to randomly divide 192 patients of periarthritis of shoulder into 8 groups, 24 cases in each one. Separately, 4 factors and each different 2 levels were adopted in treatment, named acupuncture timing (factor A: A, acute stage, A2 adhesion stage), acupoint combination (factor B: B, local acupoints, B2 local acupoints and distal acupoints along meridians), filiform needling and warm needling therapy (factor C: C1 acupuncture with filiform needle, CZ acupuncture with filiform needle and warm needling therapy) and positive functional exercise (factor D: D1 without positive functional exercise, D2 with positive functional exercise). The treatment was given once a day, 10 treatments made one session and 2 sessions were required totally. The time points of observation were the point after 1 session of treatment and after 2 sessions of treatment. The short-form McGill pain questionnaire (MPQ) and shoulder joint motor disturbance score were adopted for evaluation.</p><p><b>RESULTS</b>In the orthogonal design analysis, taking the hierarchical factors into consideration, the age was considered as the main factor in the evaluation of shoulder pain and shoulder motor disturbance (P<0.01), and the shoulder function grade apparently impacted pain evaluation and the efficacy on shoulder motor disturbance (P<0.01). The best combination of 4 factors and 2 levels were A1B1CzD2 and A2BC2D2. SAS statistical analysis showed that at acute stage and adhesion stage, CZ Dz , meaning acupuncture with fifiform needling and warm needling therapy combined with positive functional exercise, is the main factor of the improvements of shoulder motor function (P<0.05).</p><p><b>CONCLUSION</b>For periarthritis of shoulder at acute stage, the combined therapy of acupuncture at local acupoints, warm needling and positive functional exercise is adopted. At chronic stage, the combined therapy of acupuncture at local acupoints and distal acupoints, acupuncture with filiform needle and warm needling and positive functional exercise is the best program. Additionally, in clinical treatment, the patients' age, sex, shoulder joint function and duration of treatment should be considered comprehensively for the impacts on the efficacy.</p>

Biological characteristics of calliphoridae and its application in forensic medicine / 法医学杂志

Diptera Calliphoridae is the first major kind of flies that appears on the decomposed corpses. In forensic entomology, according to the living characteristics of Calliphoridae flies, we could accurately estimate postmortem interval (PMI) in a murder or unidentified case and could provide useful clues to solve the case. This paper introduces the characteristics of the biology and morphology of Diptera Calliphoridae, and reviews the combined application of forensic entomology, molecular biology, mathematical morphology and toxicology.

Influence of masticatory fatigue on the fracture resistance of the pulpless teeth restored with quartz-fiber post-core and crown / 国际口腔科学杂志·英文版

To investigate whether masticatory fatigue affects the fracture resistance and pattern of lower premolars restored with quartz-fiber post-core and full crown, 44 single rooted lower premolars recently extracted from orthodontic patients were divided into two groups of 22 each. The crowns of all teeth were removed and endodontically treated and then restored with quartz-fiber post-core and full crown. Twenty-two teeth in one group were selected randomly and circularly loaded at 45° to the long axis of the teeth of 127.4 N at a 6 Hz frequency, and the other group was not delivered to cyclic loading and considered as control. Subsequently, all teeth in two groups were continually loaded to fail at 45° to the long axis of the teeth at a crosshead speed of 1 mm⋅min(-1). The mean destructive force values were (733.88±254.99) and (869.14±280.26) N for the experimental and the control group, respectively, and no statistically significant differences were found between two groups (P>0.05). Bevel fracture and horizontal fracture in the neck of root were the major fracture mode of the specimens. Under the circumstances of this study, it seems that cyclic loading does not affect the fracture strength and pattern of the quartz-fiber post-core-crown complex.

Curcumin induces apoptosis through the mitochondria-mediated apoptotic pathway in HT-29 cells / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

<p><b>OBJECTIVE</b>To investigate the effects of curcumin on release of cytochrome c and expressions of Bcl-2, Bax, Bad, Bcl-xL, caspase-3, poly ADP-ribose polymerase (PARP), and survivin of HT-29 cells.</p><p><b>METHODS</b>HT-29 cells were treated with curcumin (0 approximately 80 micromol/L) for 24 h. The release of cytochrome c from the mitochondria and the apoptosis-related proteins Bax, Bcl-2, Bcl-xL, Bad, caspase-3, PARP, and survivin were determined by Western blot analysis and their mRNA expressions by reverse transcriptase-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>Curcumin significantly induced the growth inhibition and apoptosis of HT-29 cells. A decrease in expressions of Bcl-2, Bcl-xL and survivin was observed after exposure to 10 approximately 80 micromol/L curcumin, while the levels of Bax and Bad increased in the curcumin-treated cells. Curcumin also induced the release of cytochrome c, the activation of caspase-3, and the cleavage of PARP in a dose-dependent manner.</p><p><b>CONCLUSION</b>These data suggest that curcumin induced the HT-29 cell apoptosis possibly via the mitochondria-mediated pathway.</p>

Evaluation of clinical performance of cardiac troponin Ⅰ in diagnosis and prognosis of non-ST elevation acute coronary syndromes / 中华检验医学杂志

Objective To evaluate the sensitivity and specificity of new genevation Beckman Coulter cTnI in diagnosis of non-ST-elevation myocardial infarction(NSTEMI),and the values of cTnI in prognosis of non-ST elevation acute coronary syndromes(NSTEACS).Methods The cTnI was determined in NSTEMI patients and in control group,then analyzed the data with receiver operating characteristic curve (ROC curve)statistical software.Two hundred and sixty-nine patients with non-ST elevation acute coronary syndromes were divided into cTnI≥0.04 ?g/L and cTnI