The training of teaching ability is an important context of master course.Pathology Department of Xiang Ya School of Medicine take experimental teaching practice as the important way to improve their pathological experimental teaching skill.In this paper we introduced our experience.

Experimental teaching plays an important role in pathological teaching.We have taken effective measures to renovate the methods for experimental teaching,establishing open teaching mode.As a result,teaching quality increases and students become more active in pathological learning.

BACKGROUND: Carbonated hydroxyapatite cement (CHC) s a new kind of biomaterial for bone defect, which is made of powder and fluid, and can be mixed to be pasty to repair various bone defects.OBJECTIVE: To observe the improvement of vertebrae height and pain in patients with osteoporosis vertebral compression fracture (VCF) after vertebroplasty by using a new kind of bone graft biomaterial, taking CHC as the filling material to reinforce the vertebral body.DESIGN: A contrast observation trial taking patients as subjects.SETTING: Department of Orthopaedics, General Hospital of Chinese PLA.PARTICIPANTS: Totally 34 patients with thoracic or lumbar osteoporosis VCF who received the treatment in the Department of Orthopaedics, General Hospital of Chinese PLA between October 2000 and August 2003. Inclusive criteria: ①Definite diagnosis by CT; ② Informed consents were obtained from the patients. Exclusive criteria: The patients with osteoporosis vertebral compression fractures who suffered vertebral posterior wall fracture. There were 6 males and 28 females, and they were aged (72±13)years; Among the patients, 27 were diagnosed as postmenopausal osteoporosis, 1 as cortical hormone-induced osteoporosis and 6 male patients weresenile osteoporosis.METHODS: ①All the patients were randomly divided into two groups: Experimental group (n =23) and control group (n=11). All the patients were performed percutaneous operation with local anesthenia. All cases were performed percutaneous operation under local anesthesia. Under the C-arm monitored, one side pedicle puncture was performed to enter the anterior column of the involved VCF. Patients of the experimental group were filled with CHC. Patients of control group were filled with polymethyl Methacrylate (PMMA) with the same way. ② Referred to McGill-Melzack scoring. Among the scale 0-100 mm (0 was no pain, 100 was acute pain), the value indicated the painful intensity and mental assault degree. ＜ 30 scores indicated good, 30-40 basically satisfied and ≥ 50 poor .③ Referred to the method from Lee et al, the preoperative height (A1) and postoperative height (A2) of compression fracture position of VCF were measured according to the lateral X-ray film. At the same time, the upper vertebral height (A3) and the inferior vertebral height (A4) were measured at the same position. The original height (A) of the involved vertebra was calculated as (A)= (A3+A4)/2,and the preoperative vertebral compression rate =(A-A1 )/A, the postoperative vertebral compression rate =(A-A2)/A, the restoring rate = (the preoperative vertebral compression rate-the postoperative vertebral compression rate)/the preoperative vertebral compression rate. ④ The wounds of the patients were observed after operation. The levels of blood routine, serum calcium and serum phosphorus were detected before, one day and one week after operation. MAIN OUTCOME MEASURES: ① Preoperative and postoperative VAS scoring. ② The vertebral compression rate and restoring rate. ③ Wounds were observed after operation. The blood routine, the serum calcium and serum phosphorus were detected before, one day and one week after operation.RESULTS: Totally 34 patients were involved in the result analysis. ①The preoperative visual analogue scale (VAS) score of experimental group were (91.5±21.7) points, and the postoperative ones were (44.5±27.2) points. The difference of VAS score reduced gradually along with the postoperative time. There was no difference of VAS score between experimental group and the control group 4 weeks after operation. ② The biocompatibility of CHC in the vertebral body was fine. The vertebral compression rate of experimental group was recovered from (43.1±21.4)％ preoperatively to (27.3± 18.5)％ postoperatively. The rate of restored heights was (27.3±18.5)％. ③ All patients obtained Ⅰ stage wound healing, and none of them had infection, inflammatory secretion and nervous symptom. There were no differences in blood routine test, serum calcium, serum phosphorus between patients in two groups. One case filled by PMMA and two cases filled by CHC presented leakage, and none had nervous symptom.CONCLUSION: As the filling materials for vertebropalsty, CHC can restore the vertebral heights and relieve pain safely and effectively, however, its efficacy to relieve pain is not significant as PMMA in the short term.

Objective To observe the role and related mechanism of chemerin and its receptor ChemR23 in glomerular endothelial cells (GEnCs) stimulated by high glucose.Methods Mouse GEnCs were cultured and divided into control group,20.0 mmol/L high glucose group,40.0 mmol/L high glucose group and mannitol control group.Then the expressions of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in cell culture supematant as well as the expressions of intracellular protein and mRNA of chemerin,ChemR23,IL-6 and TNF-α were detected.Lentiviral transfection targeting ChemR23 was applied before high glucose-or Chemerin-stimulated,and expressions of supernatant and intracellular mRNA of IL-6 and TNF-α were measured.Meanwhile whether p38 mitogen-activated protein kinase (p38 MAPK) pathway was activated by high glucose was detected.The specific inhibitor of p38 MAPK was added prior to high glucose-stimulated,then supernatant and intracellular mRNA expressions of IL-6 and TNF-α was detected.The supernatant expressions of IL-6 and TNF-α were measured by ELISA.The intracellular protein expression and p38 MAPK phosphorylation activity were detected by Western blotting.The mRNA expression was detected by real time PCR.Results Compared with those in the control group,in high glucose groups the expressions of IL-6,TNF-α and chemerin were significantly increased (all P ＜ 0.05),however,the expressions of ChemR23 did not change (all P ＞ 0.05);the supernatant and mRNA expressions of IL-6 and TNF-α were also elevated in the chemerin group (all P ＜ 0.05).Lentivirus baring shRNA could efficiently suppress ChemR23 expression,and the Chemerin-or high glucose-induced expressions of IL-6 and TNF-α were reduced (all P ＜ 0.05).Also it could significantly reduce the expression of phosphorylated-p38 MAPK (p-p38 MAPK) induced by high glucose (P ＜ 0.05),as high glucose group had higher p-p38 MAPK than control group (P ＜ 0.05).While the high glucose-elevated expressions of IL-6 and TNF-α were significantly attenuated by p38 MAPK inhibitor (all P ＜ 0.05).Conclusions High glucose stimulation can induce the expression of chemerin in GEnCs.By binding to ChemR23,chemerin activates p38 MAPK signaling pathway,and then promotes the expressions of IL-6 and TNF-α.These inflammatory cytokines aggravate inflammation of GEnCs.

Long-term medical education program requires that the medical students should ac-quire both professional knowledge and scientific research ability. These students,with heavy task and course,have difficulty in performing the scientific research systematically. It is very important to develop the early scientific research training. Department of pathology in Central South University,took early sci-entific research activities in various forms,such as literature searching,reviews writing,research design-ing,experiment performing,lecture communicating and clinical practicing after combining the discipline characteristic and arranging the overall process. Satisfactory effects were achieved with efforts.

Objective To investigate the effect of knockdown or overexpression of G6PD on proliferation, growth and migration of human hepatocellular carcinoma cell PLC/PRF/5. Methods Lentivirus-mediated knock-down or overexpression of G6PD was achieved in human hepatocellular carcinoma cell line PLC/PRF/5. RT-PCR and Western blotting assay were used to detect the overexpression or knockdown of G6PD.Cell proliferation and mi-gration curves were recorded by real-time cell analysis system(RTCA),the cell proportion in the DNA replication phase can be directly displayed with EDU experiment,cell growth ability was detected by colony forming assay. Results The doubling time of cells in G6PD knockdown group was longer than that of the control group,and the cell growth rate decreased significantly,the proportion of cells in proliferative phase(43.2%)was lower than that in the control group,but the rates colony formation and migration were significantly decreased(P<0.05,respective-ly),and the migration curves separated apparently.While no significant differences in proliferation,growth and mi-gration of PLC/PRF/5 cells were found between the over-expressed strain and the control group. Conclusion The reduction of G6PD expression in HCC cells inhibits the proliferation and growth of HCC,which may lay a foun-dation for the further study of the pathogenesis and treatment of HCC.

ABSTRACT:Objective To investigate the effect of PDCA nursing model in children with mycoplasma pneumonia.Methods 120 children with mycoplasma pneumonia treated with Tanre-qing combined with azithromycin were randomly divided into the observation group (PDCA nursing model)and the control group (usual care)with 60 cases in each group.Clinical compliance,effica-cy,clinical symptoms,hospitalization time and adverse reactions were compared in two groups.Re-sults The clinical compliance in the observation group was higher than that in the control group(P<0.05).The effective rate in the observation group was 98.33%,which was significantly higher than that in the control group(88.33%)(P <0.05).Cooling time observation group,cough time, rales disappeared time,tonsils congestion disappear time and length of hospital stay in the observa-tion group were significantly less than those in the control group (P <0.05).Incidence of adverse reactions in the observation group was 15.00%,which was significantly lower than that in the con-trol group(31.67%)(P <0.05).Conclusion PDCA nursing model significantly improves compli-ance and clinical efficacy in children with mycoplasma pneumonia,and shortens the time of clinical symptoms and hospitalization and reduces adverse side effects.

ABSTRACT:Objective To investigate the effect of PDCA nursing model in children with mycoplasma pneumonia.Methods 120 children with mycoplasma pneumonia treated with Tanre-qing combined with azithromycin were randomly divided into the observation group (PDCA nursing model)and the control group (usual care)with 60 cases in each group.Clinical compliance,effica-cy,clinical symptoms,hospitalization time and adverse reactions were compared in two groups.Re-sults The clinical compliance in the observation group was higher than that in the control group(P<0.05).The effective rate in the observation group was 98.33%,which was significantly higher than that in the control group(88.33%)(P <0.05).Cooling time observation group,cough time, rales disappeared time,tonsils congestion disappear time and length of hospital stay in the observa-tion group were significantly less than those in the control group (P <0.05).Incidence of adverse reactions in the observation group was 15.00%,which was significantly lower than that in the con-trol group(31.67%)(P <0.05).Conclusion PDCA nursing model significantly improves compli-ance and clinical efficacy in children with mycoplasma pneumonia,and shortens the time of clinical symptoms and hospitalization and reduces adverse side effects.

Objective To observe the expression of ChemR23 induced by Angiotensin Ⅱ (Ang Ⅱ) in podocyte and its role in renal injury.Methods Conditionally immortalized mice podocytes were cultured in vitro.Immunofluorescence was used to observe the sub-cellular location of ChemR23.The expressions of ChemR23,Nephrin and Podocin stimulated by different concentrations of Ang Ⅱ were detected by qRT-PCR and Western blotting.Lentivirus targeting ChemR23 was used.The expressions of Nephrin and Podocin and the phosphorylation state of NF-κB P65 were detected by Western Blot.The inhibitor of NF-κB P65 was added to the cultural medium for 2 h before Ang Ⅱ stimulation.The effect of NF-κB P65 inhibitor on Ang Ⅱ-induced expression of Nephrin and Podocin was detected by Western Blot.Results It is showed that ChemR23 was located in cytosol and membrane.Compared with the normal control,the expression of ChemR23 was significantly increased by Ang Ⅱ in mRNA and protein level,while the expressions of Nephrin and Podocin were decreased (P ＜ 0.05).When using Lentivirus vector to interfere the expression of ChemR23,Ang Ⅱ-repressed expressions of Nephrin and Podocin were restored (P ＜ 0.05).Western Blot showed the level of phosphorylated NF-κB P65 was significantly increased by Ang Ⅱ stimulation (P ＜ 0.05),which could be inhibited by interfering the expression of ChemR23.When adding the NF-κB P65 inhibitor,the low expression of Nephrin and Podocin induced by Ang Ⅱ stimulation was restored (P＜0.05).Conclusions Ang Ⅱ can induce ChemR23 expression,which activates NF-κB P65 signaling pathway,and then inhibits the expressions of Nephrin and Podocin.Targeting ChemR23 is a potential way to alleviate podocyte injury caused by Ang Ⅱ.

OBJECTIVETo detect pathogenic mutations in Marfan syndrome (MFS) using an Ion Torrent Personal Genome Machine (PGM) and to validate the result of targeted next-generation semiconductor sequencing for the diagnosis of genetic disorders.

METHODSPeripheral blood samples were collected from three MFS patients and a normal control with informed consent. Genomic DNA was isolated by standard method and then subjected to targeted sequencing using an Ion Ampliseq(TM) Inherited Disease Panel. Three multiplex PCR reactions were carried out to amplify the coding exons of 328 genes including FBN1, TGFBR1 and TGFBR2. DNA fragments from different samples were ligated with barcoded sequencing adaptors. Template preparation and emulsion PCR, and Ion Sphere Particles enrichment were carried out using an Ion One Touch system. The ion sphere particles were sequenced on a 318 chip using the PGM platform. Data from the PGM runs were processed using an Ion Torrent Suite 3.2 software to generate sequence reads. After sequence alignment and extraction of SNPs and indels, all the variants were filtered against dbSNP137. DNA sequences were visualized with an Integrated Genomics Viewer. The most likely disease-causing variants were analyzed by Sanger sequencing.

RESULTSThe PGM sequencing has yielded an output of 855.80 Mb, with a > 100 × median sequencing depth and a coverage of > 98% for the targeted regions in all the four samples. After data analysis and database filtering, one known missense mutation (p.E1811K) and two novel premature termination mutations (p.E2264X and p.L871FfsX23) in the FBN1 gene were identified in the three MFS patients. All mutations were verified by conventional Sanger sequencing.

CONCLUSIONPathogenic FBN1 mutations have been identified in all patients with MFS, indicating that the targeted next-generation sequencing on the PGM sequencers can be applied for accurate and high-throughput testing of genetic disorders.

Base Sequence ; Computational Biology ; Fibrillin-1 ; Fibrillins ; Genomics ; High-Throughput Nucleotide Sequencing ; methods ; Humans ; Marfan Syndrome ; diagnosis ; genetics ; Microfilament Proteins ; genetics ; Mutation ; Semiconductors