Under the guidance of the learning theory of constructivism,the mode of curriculum educational reform is designed and experimented on the"acupuncturology" on-line study website using the method of action research.The action experiment contains 3 kinds of different teaching modes: the teaching center mode,the study center mode and teacher donimated-students centered mode.A whole process intervention to question senses is carried on.The relative theoretical basis and problem-solving research methods are found out during the experiment.Phase summary is formed and the teaching project is constantly revised and become perfect.

BACKGROUND:Fracture healing is the coupling interaction of osteoblasts and osteoclasts that promotes bone growth, in which osteoblast-mediated bone resorption and osteoclasts-mediated bone reconstruction make the bone reconstruction in a dynamic equilibrium to promote bone growth. However, the majorities of the current studies focus on osteogenic or osteoclastic mechanism alone, and ignore the interaction mechanism between these two cels under co-existing conditions.
OBJECTIVE:To investigate the effects of Kidney Chinese Herbs on osteoblasts and osteoclasts coupling of osteoprotegerin-receptor activator of nuclear factor kappaB ligand-receptor activator of nuclear factor kappaB and its mechanism of action in fracture treatment.
METHODS: Mouse osteoblasts and osteoclasts were isolated and cultured in vitro to establish the mouse “osteoblast-osteoclast co-culture system” as a research platform. Then, Kidney Chinese Herbs at doses of 1.25, 2.5, 6.25 g/(kg?d) were given intragastricaly for 7 consecutive days. Mice in the blank control group were fed with the same volume of normal saline.
RESULTS AND CONCLUSION:The alkaline phosphatase activity in osteoblasts co-cultured with osteoclasts was significantly higher than that in osteoblasts cultured alone at 24 hours of culture (P < 0.05). Real-time PCR showed that in the co-culture system, the expression of alkaline phosphatase, Runt related transcription factor 2 and osteoprotegerin were increased in a dose-dependent manner (P < 0.05). Western blot assay showed 6.25 g/(kg?d) Kidney Chinese Herbs could dramaticaly promote the expression of osteoprotegerin and receptor activator of nuclear factor kappaB ligand, but restrained the expression of receptor activator of nuclear factor kappaB (P < 0.05). These findings indicate that Kidney Chinese Herbs can dynamicaly regulate the osteoprotegerin-receptor activator of nuclear factor kappaB ligand-receptor activator signaling pathway, and has a positive effect to promote bone reconstruction and rehabilitation.

Objective To study the clinical rescue for basicranial fracture complicated with massive hemorrhage.Methods The therapies for 20 patients of basicranial fracture complicated with massive hemorrhage treated in our hospital within 3 years were analyzed retrospectively.Results Among 20 patients,16 cases were male,4 cases were female.7 cases were simple anterior basicranial fracture with massive hemorrhage,1 case was the delayed massive hemorrhage in cavernous fistula caused by anterior basicranial fracture,12 cases were anterior and mid basicranial fracture complicated with massive hemorrhage.In consequence,6 cases were secondary cerebrospinal leak,8 cases recovered,8 cases died.Conclusion The basicranial fracture complicated with massive hemorrhage is a very dangerous symptom with high death rate and high disability rate.The patients should be rescued actively.

[Objective] To investigate the effects of the bioactive constituents of green tea polyphenols epigallocatechin gallate (EGCG) on low density lipoprotein receptor (LDLR) function of HepG2 cells.[Methods] The optimal concentration and cell proliferation of HepG2 cells were determined by CCK8 assay,and Western blotting was used to determine LDLR and PCSK9 protein levels,respectively,and LDL uptake in HepG2 cells was detected by fluorescence microscope.[Results] EGCG elevated LDLR protein expression,reduced PCSK9 protein expression and promoted LDL uptake in HepG2 cells.[Conclusion] EGCG may increase LDLR abundance by down-regulating PCSK9 protein and attenuating LDLR protein degradation,which providing a new approach for lipid lowering therapy.

ObjectiveTo analyze the relationship between pancreatic cancer (PC) and diabetes mellitus (DM),and the clinical and pathological features of pancreatic cancer in patients with DM.MethodsFrom January,2008 to December,2010,151 patients with PC and 195 comparable patients without PC were enrolled in a case-control study to analyze the relationship between PC and DM.ResultsThe OR was 5.91 (95％ CI 3.03-8.00,P＜0.05) in PC patients with DM for less than 2 years,and 1.308 (95％ CI 0.37-4.60,P＞0.05) in patients with DM for 2 to 5 years,and 1.16(95％ CI 0.44-3.19,P＞0.05) for patients with DM for more than 5 years.There was no significant difference between PC patients with and without DM in gender,age,body-mass index (BMD,obstructive jaundice,tumour location and tumour metastases (P＞0.05).ConclusionsThere was a significant correlation between PC and DM.De novo DM may be a clinical manifestation in patients with PC.PC patients with concurrent DM have no particular clinical and pathological features.

[ ABSTRACT] AIM:To investigate the effects of induced pluripotent stem cells-derived mesenchymal stem cells ( iPSC-MSCs) on cobalt chloride ( CoCl2 )-induced injuries of PC12 cells and its possible mechanism.METHODS:PC12 cells were exposed to CoCl2 to set up a chemical-induced cellular injury model and were cocultured with iPSC-MSCs.The cell viability was tested by CCK-8 assay.The apoptosis was measured by flow cytometry using Annexin V/PI staining.The mitochondrial membrane potential (MMP) was analyzed by flow cytometry using JC-1 staining.Immunofluorescence was employed to observe mitochondrial transfer from iPSC-MSCs to PC12 cells.RESULTS: Apoptosis of PC12 cells was in-creased and MMP of PC12 cells was decreased after exposed to CoCl2 at concentration of 400μmol/L for 24 h.Coculture of PC12 cells with iPSC-MSCs reduced the apoptosis and recovered the MMP of the PC12 cells.Tunneling nanotubes were formed between iPSC-MSCs and PC12 cells, through which the iPSC-MSCs transferred the mitochondria to the PC12 cells. CONCLUSION:iPSC-MSCs protect PC12 cells from CoCl2-induced injuries, which may be associated with the mitochon-drial transfer from iPSC-MSCs to PC12 cells.

AIM: To explore the effect and possible mechanism of type 2 innate lymphoid cell (ILC2) on the development of chronic renal failure (CRF).METHODS: The patients with chronic renal failure (n=36) in the Fist Affiliated Hospital of Sun Yat-sen University from March 2016 to December 2016 were selected, and 32 healthy persons in the same period were enrolled in the study for control.The proportion of ILC2 in the PBMC of CRF patients and healthy controls was detected by flow cytometry, IL-13 concentration in the plasma was measured by ELISA.The isolated PBMCs from the patients and healthy persons were divided into 3 groups (control group, cytokine group, intervention group) and cultured in vitro for 3 days, respespestively, then IL-13 concentration was measured by ELISA.The protein levels of phosphorylated signal transducers and activators of transcription 6 (p-STAT6) in the PBMC of healthy controls before stimulation and after stimulation for 15 min, 30 min, 1 h, 2 h were determined by Western blot.RESULTS: The proportion of ILC2 in the PBMC and the plasma IL-13 concentration of CRF patients was higher than that in the healthy controls (P<0.05).In the culture supernatant in vitro, IL-13 concentration in the 3 subgroups of CRF patients (control group, cytokine group, intervention group) were all higher than that in the healthy controls (P<0.05), both the 2 groups showed a trend that the active IL-13 concentration in cytokine group was higher than that in control group, and that in intervention group was lower than that in cytokine group.The protein levels of p-STAT6 in cytokine stimulated-PBMC with a time dependent manner.CONCLUSION: The percentage of ILC2 in the PBMC is elevated in CRF patients.Furthermore, the ILC2 secret large amount of IL-13 to mediate the polarization of Th2 cells to regulate immunity through activating p-STAT6.

Objective To investigate the consistency of the results detected by different types of blood cell analyzer in the blood center .Methods On the basis of the calibration in each analyzer ,with the analyzer obtaining the excellent result in participating the external laboratory quality assessment hosted by the Ministry of Health as the reference ,the fresh blood samples were adopted to analyze other analyzers .Results For the analyzers after conducting the comparison ,the consistency and accuracy of the detection results were ensured .Conclusion After calibration in different types of blood cell analyzer ,the differences exist in the detection re‐sults .Periodically conducting the comparison among different types of instrument has very practical significance .

Objective To evaluate the quality of pooled platelets leukocytes reduced after filtering out leukocytes using two man-ufacturers of leucocyte filters for pooled platelets and improving the preparation method.Methods Pooled platelets was prepared from 400 mL fresh whole blood by buffy coats(BC)method,after 1 6 h,(22±2)℃ holding period,pooled six bags of ABO-matched buffy coats.and then filtered with two manufacturers of leucocyte filter,divided into the control group and the experimental group. Before and after filtering,the numbers of platelet and leukocyte,pH,hypotonic shock response(HSR),platelet aggregation and CD62p expression were detected.Results Before filtering leukocytes,the platelet quality of two groups achieved the requirements of Chinese standards.The numbers of platelet and leukocyte,pH,CD62p expression(%)and platelet aggregation showed no signifi-cant difference between two groups(P >0.05).However,After filtering,the pH,platelet aggregation and the platelet recovery,the experimental group and the control group,were (6.53±0.60)vs(7.00±0.06)、(5.5±3.8)% vs (77.4±14.7)%,(86.8±4.3)%vs (90.6±2.7)%,showed significant differences (P <0.05);but the residual leukocytes,HSR and CD62p expression respectively were (3.00±4.00)×10 6/ bag vs (2.00±3.00)×10 6/ bag,(2.40±0.90)% vs (2.00±0.80)%,(7.30±5.90)% vs (5.60 ±3. 70)%,there were no significant differences(P >0.05).Conclusion The quality of pooled platelets leukocytes is reduced,after filte-ring out leukocytes with two manufacturers of leucocyte filters and improving the preparation method,achieves the requirements of Chinese standards.However,the leukocyte filters of experimental group might have influence on platelet aggregation and pH.

Objective To establish the cardiac arrest-cardiopulmonary resuscitation model in rats, and to observe the effect of mild hypothermia on autophagy in hippocampal CA1 neurons after ROSC.Methods A total of 36 Wistar rats were randomly divided into two groups: normal temperature treatment group(NT group) and mild hypothermia treatment group(HT group).To establish the cardiac arrest-cardiopulmonary resuscitation(CA-CPR) model in rats by epicardial electrical stimulation induced ventricular fibrillation, and to sacrifice 3 animals in each group to obtain the brain cortex in 2nd and 4th hours after ROSC in order to observe the expression of p-AMPK by electron microscope and LC3 granules through Western blot.The neurological deficit score(NDS) was assessed in 24、48、72 hours respectively after ROSC.To sacrifice the animals so as to take the cerebrum in 72 hours after ROSC, then calculate the apoptotic index of the hippocampal CA1 neurons, which were dyed through TUNEL method.Results The expression of p-AMPK、Beclin-1 and LC3-Ⅱ/LC3-Ⅰratio in Normothermia group were all lower than the Mild hypothermia group(P<0.05), the neurons plasma of hippocampal CA1 area in the Hypothermia group demonstrated obvious LC3 granules formation, the NDS score of the Normothermia group and the Mild hypothermia group in ROSC24h、ROSC48h、ROSC72h were 320vs205、285vs140、266vs120, respectively.The apoptotic index of the hippocampal CA1 area in the Normothemia group in ROSC72h was higher than the Mild hypothermia group,(P<0.05).Conclusions Mild hypothermia after cardiopulmonary resuscitation promotes autophagy of the hippocampal CA1 area neurons in rats and reduce neuronal apoptosis.