Functional Science Lab.Of Jining Medical College has attempted to reform the management system,share the resource,improve the resource utilization,exceed the limits of science,modify the contents of experimental courses,innovate the teaching methods,pay attention to teacher training.Through longtime research and practice,we have gained the significant achievements.

Objective To study a scoring method for assessing function status of cardiovascular system quantitatively. Methods First, exercise heart rate variability (EHRV) was obtained from the primary dynamic electrocardiogram (ECG) recorded during exercise testing. Then appropriate entropy, relative complexity, and other three parameters extracted from poincare dispersed-dot plot were extracted from the EHRV. Discriminant analysis was used to classify two extreme groups.Results Based on the values of two groups from the discriminant formula, a scoring formula was proposed and four ranks were divided according to different score domains. A novel scoring method was established. To validate the proposed scoring method, 60 middle-aged hypertensives, 50 elder subjects and 110 young healthy subjects were examined and scored. Conclusion Scores of 220 subjects are consistent with their real health status. The proposed scoring method reflects the status of the subject's cardiovascular system effectively.

Designing experimentation is the most important phase in function experimentation teaching,and a simulation process of science research.Students choose the topic freely,design schema and practice by themselves.Based on pathophysilolgy,the article has practiced functional designing experimentation among the students of clinical medicine in 2006 grade.Results indicated that the practice of designing experimentation is helpful for raising general diathesis and innovative ability of students,and it will be the focus of function experimentation innovation in future.

BACKGROUND: It has been widely accepted that both bone marrow-derived mesenchymal stem cells (MSCs) and hematopoietic stem cells (HSCs) have the capacity to differentiate into neural lineages. Some scholars believe that in addition to HSCs and MSCs, bone marrow (BM) also harbors a highly mobile population of CXCR4+ tissue committed stem cells (TCSCs), including skeletal muscles, heart, liver, and neural tissue. OBJECTIVE: To make sure that neural tissue-committed stem cells (NTCSCs) reside in the bone marrow, and to establish a purification and culture method for bone marrow-derived NTCSCs.DESIGN: Opening animal study.SETTING: Department of Anatomy, the Second Military Medical University of Chinese PLA. MATERIALS: Adult Sprague-Dawley (SD) rats (pathogen-free) were provided by the Animal Center of the Second Military Medical University of Chinese PLA. Dulbecco's modified eagle's medium (DMEM)/F12, B27, N2 and epidermal growth factor (EGF, Invitrogen Company), basic fibroblast growth factor (bFGF, CytoLab Ltd), rabbit anti-rat Nestin,CXCR4, β-Tublin Ⅲ, glial fibrillary acidic protein (GFAP, Santa Cruz Company), mouse anti-rat microtubule associated protein 2ab (MAP2ab) (Clone11-5B), cyclic nucleotide 3'phosphohydrolase (CNPase, Clone AP20, NeoMarkers Company), fluorescent(fluorescein isothiocyanate, Cy3) marker reagents (Wuhan Boster Bioengineering Co., Ltd), nuclear fluorescent dyes 4,6-diamidino-2-phenylindole(DAPI)(Sigma), immunohistochemistry reagents (Vector Laboratories Company) , and NycoPrepTM separation liquid (1.077A, Axis-Shield Company) were used in this study.METHODS: This study was performed in the Department of Anatomy, the Second Military Medical University of Chinese PLA from January 2004 to December 2006. Bone marrow was harvested from bilateral femurs and tibias of 2-3 weeks SD rats. Mononuclear cell layer was isolated by NycoPrepTM separation liquid and suspended in DMEM/F12(1:1)serum-free medium supplemented with 2% B27,1% N2, 20 μg/L bFGF, 20μg/L EGF, 1×105 U/L penicillin and 100 mg/L streptomycin. NTCSCs were isolated and propagated by suspensive growing from adherent cells in bone marrow in DMEM/F12 free-serum medium. MAIN OUTCOME MEASURES: NTCSCs were identified by immunocytochemistry for CXCR4, a marker of TCSCs and nestin, a marker of neural stem cells, and neural lineages marker protein after differentiation of cellular spheres. RESULTS: The NTCSCs spheres expressed nestin, a neural stem cell marker as well as CXCR4, a marker of TCSCs. The NTCSCs' spheres were naturally differentiated in DMEM medium with 15% fetal bovine serum. The differentiated cells expressed β-Tublin Ⅲ, MAP2ab, CNPase and GFAP, markers of neural lineages. CONCLUSION: NTCSCs reside in bone marrow and naturally differentiate into neural lineages in vitro.

BACKGROUND: It is proved that acupuncture can remarkably promote recovery of nervous function after spinal cord injury (SCI). Previous studies in our groups have been proved that electro-acupuncture can inhibit apoptosis in early period of SCI, but the mechanism is unclear yet.OBJECTIVE: To study the effect of electro-acupuncture on expressions of apoptosis inhibitory gene Bcl-2 mRNA and protein with hybridization in situ and immunohistochemistry and discuss the possible mechanism of apoptosis inhibited by electro-acupuncture in early SCI.DESIGN:Opening animal study.SETTING: Department of Anatomy, the Second Military Medical University of Chinese PLA; Shanghai Acupuncture & Moxibustion and Meridian Research Center.MATERIALS:Adult male SD rats of pathogen-free aged 2-3 months were selected in this study. Bcl-2 hybridization in situ kit was provided by Wuhan Boshide Biotechnology Company Limited and Bcl-2 antibody (1:200) was provided by Santa Cruz Biotechnology Company.METHODS: The experiment was completed at the Laboratory of Anatomy of the Second Military Medical University of Chinese PLA from July 2004 to December 2005. All experimental rats were randomly divided into model group, electro-acupuncture group, methylprednisolone group and sham operation group. T10 spinal cord was injuried by the modified Allen's method and treated with electro-acupuncture immediately, and then the expressions of Bcl-2 mRNA and protein were evaluated with hybridization in situ and immunohistochemistry combined with image quantitative analysis.MAIN OUTCOME MEASURES: ① Expressions of Bcl-2 mRNA and protein in early SCI; ② effect of electro-acupuncture on expressions of Bcl-2 mRNA and protein.RESULTS: The rats were supplied when they died during the experiment,and all 42 rats were involved in the final analysis. ① Moderate expression of Bcl-2 mRNA and protein was observed in the sham operation group.Expression of Bcl-2 mRNA was increased in model group at 6 hours after SCI, but expression ofBcl-2 protein was not changed. At 24 hours after SCI, both expressions of Bcl-2 mRNA and Bcl-2 protein were increased.Expression of Bcl-2 mRNA and protein was higher in electro-acupuncture group than that in mo del group (P ＜ 0.05), and there was no significant difference from that in methylprednisolone group. ② Amount of positive Bcl-2 mRNA cells was increased in electro-acupuncture group and methylprednisolone group at 6 hours after treatment, and gray value was decreased.There was significant and remarkably significant difference from those in sham operation group and model group, respectively (P ＜ 0.05-0.01). After 24-hour treatment, amount of positive Bcl-2 mRNA cells was increased in electro-acupuncture group and gray value was decreased.There was significant and remarkably significant difference from those in sham operation group and model group, respectively (P ＜ 0.05-0.01). ③ After 24-hour treatment, amount of immunohistochenistry positive Bcl-2 cells was increased in electro-acupuncture group and gray value was decreased. There was significant and remarkably significant difference from those in sham operation group and model group, respectively (P ＜ 0.05-0.01).CONCLUSION: Electro-acupuncture can up-regulate the expressions of Bcl-2 mRNA and protein so as to inhibit apoptosis in early SCI.

Objective:To study the effects of expansion of ropivacaine hydrochloride liquid on the postoperative analgesia,sedation, hemodynamic effects and adverse drug reactions of the patients underwent endoscopic thyroidectomy,and to explore its clinical effectiveness and safety. Methods:Forty patients who were to undergo endoscopic thyroidectomy via chest-breast approach were randomly assigned to saline solution group and expansion liquid of ropivacine hydrochloride group (ropivacaine group,n = 20).The mean arterial pressure (MAP )and heart rate (HR)were also evaluated at the following time points:preoperative evaluation (T0 ),before local anesthetic infiltration (T1 ),at the end of the operation (T2 );the blood loss,the operation time,the postoperative drainage,and the hospital duration of the patients in two groups were recorded; the nausea and vomiting,the respiratory depression,the toxicity response,the infection,and the recurrent laryngeal nerve injury during 24 h after operation were recorded.The postoperative pain scores were assessed at extubaltion (T3 ),30 min (T4 ),2 h (T5 ),6 h (T6 ),12 h (T7 )and 24 h (T8 )after extubation using Visual Analog Scale (VAS).The postoperative Ramsay scores were assessed at T3 and T4 .Results:There were no significant differences in MAP and HR at T0 and T1 of the patients between two groups (P >0.05).Compared with saline solution group,the MAP and HR at T2 and T3 of the patients in ropivacaine group were significantly decreased (P < 0.05).There were significant differences in the blood loss,the operation time, the postoperative drainage,and the hospital duration of the patients between two groups (P <0.05).The number of patients required postoperative analgesia and the number patients with nausea and vomiting in ropivacaine group were decreased compared with saline solution group (P <0.05).Compared with saline group,the VAS scores at T3 ,T4 ,T5 ,T6 ,T7 and T8 of the patients in ropivacaine group were significantly decreased (P <0.05);the Ramsay scores at T3 and T4 of the patients in ropivacaine group were also decreased significantly (P <0.05).Conclusion:Expansion liquid of ropivacaine hydrochloride can reduce the stress response during endoscopic thyroidectomy effectively and reduce the postoperative pain and injury induced by operation.

[Objective] Expressions of human epidermal growth factor receptor 2(HER-2),p53,estrogen receptor (ER),and progesterone receptor (PR) in tissues of breast invasive ductal carcinoma are not only applied to determine the therapeutic regimen,but they may also be related to the prognosis.We investigated the levels of these proteins among different clinical stages and the correlations.[Method] One hundred and thirty-eight tissues from cases with breast invasive ductal carcinoma were tested with immunohistochemistry.New scoring standards and rank test were applied.The indices were digitalized and semi-quantified.[Results] In the tissues from high clinical stage,the expression of HER2 was significantly increased,while expression of PR was markedly decreased.[Conclusion] Expression of HER2 and PR might be better markers for predicting clinical stages and prognosis.

Objective:To observe the effect of sciatic nerve pulsed radiofrequency (PRF) on the activation levels of microglia cells and astrocytes in spinal dorsal horn in the chronic constriction injury (CCI) rat models, and to explore the relationships between the analgesic mechanism of sciatic nerve PRF and the activation levels of microglia cells and astrocytes in the spinal dorsal horn.Methods:Forty male SD rats were randomly divided into CCI shamoperation+RPF sham group (SS group) , CCI sham-operation+RPF group (SP group) , CCI+RPF sham group (CS group) , CCI+RPF group (CP group) (n=10) .PRF was applied on the sciatic nerve on the 4th day after CCI operation for 120s, with a maximum temperature of 42℃.The mechanical withdrawal thresholds (MWT) and thermal paw withdrawal latencies (TWL) were measured to evaluate mechanical hyperalgesia and thermal hyperalgesia 1dbefore operation (D0) and 1, 3, 5, 7dafter operation (D1, D3, D5, D7) .Western blotting method was used to detect the protein expression levels of ionized calcium binding adapter molecule 1 (Iba-1) and glial fibrillary acidic protein (GFAP) in the ipsilateral spinal dorsal horn of L3-5after pain behavioral test (D7) .Results:Compared with SS group, the rats in CS group after CCI had the significant behavioral changes, such as hallux valgus, lameness, toe bending, and foot raising during walking;MWT and TWL were decreased significantly (P＜0.01) ;the expression levels of Iba-1and GFAP proteins in the ipsilateral spinal dorsal horn were increased significantly (P＜0.05) .Compared with CS group, the behavioral changes of the rats in CP group (D5, D7) such as hallux valgus, lameness, toe bending, and foot raising during walking were alleviated significantly;MWT and TWL were increased significantly (P＜0.01) ;the Iba-1protein expression level in spinal dorsal horn was downregulated significantly (P＜0.05) and the GFAP protein expression level did not change significantly (P＞0.05) .Conclusion:PRF on sciatic nerve can relieve the neuropathic pain (NP) of the CCI rat models;PRF on sciatic nerve can inhibit activation of the microglia cells in the spinal dorsal horn.The analgesic mechanism of PRF on sciatic nerve may be associated with the inhibition of the activation of microglia cells in the spinal dorsal horn.

Objective To compare the efficacy of CT-guided radiofrequency thermocoagulation (RFTC) treatment through the inside and outside of the foramen rotundum for primary trigeminal maxillary neuralgia.Methods Forty-eight patients of both sexes,aged 58-75 yr,suffering from primary trigeminal neuralgia the 2nd trigeminal branch pain,scheduled for elective CT-guided trigeminal RFTC,were divided into 2 groups (n =24 each) using a random number table:inside of foramen rotundum group and outside of foramen rotundum group.The needle was inserted until the lateral 1/3 of the foramen rotundum was reached under CT guidance in inside of foramen rotundum group or until the outside of the foramen rotundum was reached in outside of foramen rotundum group.High-temperature RFTC 95 ℃ was performed for 120 s,repeating for 2-3 cycles.The current sensory threshold at different frequencies was measured at 1 day before and after operation.Numeric rating scale score was performed at 1 day,30 days,1 yr and 2 yr after operation,effective RFTC treatment was defined as numeric rating scale score≤ 1,and the condition of postoperative effective RFTC treatment was recorded.The development of adverse reactions such as facial hematoma,corneal ulcer,injuring the other branch and cerebrovascular accidents was recorded.Results Compared with outside of foramen rotundum group,the current sensory threshold on the affected side at frequency of 2 000 Hz was significantly increased at 1 day after operation,the rate of effective RFTC treatment was increased 2 yr after operation (P＜0.05),and no significant change was found in the incidence of postoperative facial hematoma in inside of foramen rotundum group (P＞0.05).Conclusion The long-term curative effect of CT-guided RFTC treatment for primary trigeminal maxillary neuralgia through the inside of the foramen rotundum is better than that through the outside of foramen rotundum.

Objective To establish a Juema minipig model of myocardial infarction, to evaluate the clinical indi?ces in the model pigs, and to explore the relationship between gene expression and metabolic decompensation. Methods 13 male Juema minipigs were randomly divided into control (Sham, n=5), myocardial infarction (MI, n=5) and normal control (for evaluating the recovery condition after surgery, n=3) groups. In the MI group, the ligation was done at the left descending coronary artery around the 1/3 distance to heart apex. Four weeks after the surgery, the cardiac function and serum biochemistry was analyzed. The histological changes and gene expression profiles in the myocardium in the peri?infarct area were exanimated. Results Ultrasonic images showed that the infarction was formed, the ejection fraction and fraction shortening were significantly reduced in the MI group ( ~32% and ~40% less than those of the sham group). Histological examination showed that myocardial fibers at the peri?infarct area were broken, dissolved, and there was con?nective tissue hyperplasia with increased neutrophil and lymphocyte infiltration. Microarray analysis revealed that two myo?cardial remodeling and pathology mediating pathways, three inflammation?related pathways, and 8 metabolic pathways ( in?cluding fatty acid, amino acid, and glucose metabolic pathways) were significantly changed. Conclusions We have suc?cessfully established a Juema minipig model of myocardial infarction. The less branches of the left descending coronary ar?tery allow us to establish a stable model by surgery with comparable characteristics in the clinic indices. The results of this study provides useful reference characteristics of an animal model with characteristic changes in the peri?infarct area.