Abstract:Objective: To enhance the efficiency of bone marrow transplantation (BMT) and to provide a simple and effective method for cryopreservation of bone marrow cells (BMCs). Methods: BMCs were cryopreservated at -80℃ and a new method for bone marrow scheduled transplantation (BMST) was studied with acute radiation disease model of Kunming mice. Results: The method for cryopreservation of BMCs at -80℃ produced the highest recovery rates for nucleated cells (89.0%±3.5%), the trypan blue exclusion (85.0%±3.6%), CFU-GM ［(108±10)/105 BMC］, CFU-S［(12±3)/105 BMC］. Using this method, lethally irradiated Kunming mice received allogenetic BMST. The survival rate of mice (30%) in BMST group transplanted with 4×105 BMCs was the same as that in once BMT group transplanted with 1×107 BMCs. And the survival rate of mice (60%) in BMST transplanted with 4×106 BMCs was significantly higher than that in once BMT. Evidence of engraftment could be detected in all groups. Conclusion: This method for cryopreservation BMCs at -80℃ is simple and effective, and these data suggest that “Niches” vacated each time could be fully used in BMST, so the transplanted BMCs could be decreased, the engraftment of stem cells could be increased, and the grafting efficiency could be enhanced. This method is a new method aiming at “both seed and soil”.

Abstract:Objective: To explore dynamical changes of nitric oxide synthase (NOS) activity in damaged spinal cord tissue at early phase of spinal cord injury and its cell derivation. Methods: NOS activity was determined by the rate of conversion of 3H-arginine to 3H-citrulline in damaged spinal cord tissue from 0 to 60 min after spinal cord compression, and immunohistochemistry was applied to study NOS derivation. Results: The NOS activity increased sharply after spinal cord compression injury, reached the highest at 5 min, and then decreased rapidly to normal level in 1 h. NOS-Ⅰ immunoreactive interneurons were identified mainly in the intermediolateral cell column, around the central canal and in dorsal and ventral horn, but there were no cells with NOS-Ⅲ immunoreaction in spinal cord tissue. Conclusion: There is a sharp transient increase in NOS activity during the first hour of spinal cord compression, the derivation is mainly interneurons in spinal cord.

Abstract:Objective: To evaluate different means in the diagnosis of cervical hyperextension injury with surgical treatment consideration. Methods: Fifty-three cases of cervical hyperextension injury were retrospectively analysed, and their injury history, clinical manifestation, X-ray films, MRI and treatment were studied. Results: Thirty-four cases were detected by means of X-ray films, accounting for 64.15%; the main pathological changes of MRI were rupture of anterior longitudinal ligament, partial or total horizontal tear of disc, disc herniation, compression and edema of spinal cord. All cases were detected with variable MRI abnormal changes, 7 cases of vertebral artery occlusion were found. The result of surgical treatment was superior to conservative treatment. Conclusion: MRI is superior to other diagnosis methods on injury detection, description of injury mechanism and injury severity, and it has already become the most valuable choice for diagnosis. The consideration of operation indication and surgical approach should be based on decompression and stability.

Abstract:Objective： To study the damaging effects of interleukin-6 (IL-6) and tumor necrosis factor (TNF) on human vascular endothelial cells (HVEC). Methods: A HVEC model was developed. After being incubated with IL-6, TNF and their antibody, HVEC morphological changes were observed, and the levels of lactate dehydrogenase (LDH) and angiotensin converting enzyme (ACE) in the culture fuild were measured. Results: TNF and IL-6 could cause enlargement in intercellular space, even the exfoliation of HVEC, moreover, ACE and LDH levels increased in a time- and dose-dependent manner. Conclusion: IL-6 and TNF have damaging effects on HVEC.

Abstract:Objectve: To study the effects of luteinizing hormone-releasing hormone (LHRH) analogue nafarelin, estradiol(E2) and tamoxifen on the growth of BCaP-37 human breast cancer cell line in vitro. Methods: BCaP-37 human breast cancer cells (2.5×104/well) were cultured in DMEM with 5%no estradiol bovine serum. The number of BCaP-37 cell was counted after 96 h culture in following conditions respectively: (1) with nafarelin at 10-5-10-9mol/L, (2) with E2 at 10-7-10-10mol/L, (3) with nafarelin or tamoxifen at 10-6-10-8mol/L stimulated by 10-8mol/L E2, (4) with epidermal growth factor (EGF) at 10ng/ml or 50ng/ml, (5) with nafarelin between 10-5mol/L and 10-7mol/L influenced by EGF at 10ng/ml. Results: Nafarelin had no inhibitory effect on growth of BCaP-37 cell cultured at 10-5-10-9mol/L. At the concentrations between 10-6and 10-8mol/L, nafarelin or tamoxifen could antagonize the cell growth stimulated by E2 (10-8mol/L) respectively, and nafarelin had no effect on growth of BCaP-37 cell inhibited by EGF (10ng/ml). Conclusion: Nafarelin has no inhibitory effect on growth of BCaP-37 human breast cancer cell in vitro, but can antagonize the cell growth stimulated by estradiol.

Abstract:Objective: To explore the effect of chronic hypoxia on the proliferation and dedifferentiation of LLC-PK1 cells. Methods: The cells were either exposed to hypoxia(3%O2) or maintained in normoxia(18%O2) followed by the assessment of ［3H］-thymidine incorporation and cell number as indices of cellular proliferation and sodium-dependent transport of glucose and a minoisobutyric acid(AIB) as indices of differentiation. The activity of protein kinase C(PKC) was determined with radionuclear technique. Results: Exposure of quiescent cultures to hypoxia for 16 h resulted in a significant increase in ［3H］-thymidine followed by a significant increase in cell number at 24 h in comparison with respective normoxic controls. Confluent cultures exposed to 72 h of hypoxia exhibited significant inhibition of α-methyl glucose and AIB uptakes when compared with their respective normoxic counterparts. Hypoxia also activated PKC at 4 h followed by a subsequent return to baseline with reactivation at 24 h which remained sustained up to 72 h, suggesting a biphasic acute and sustained activation of PKC. Furthermore, the hypoxia-induced alterations in ［3H］-thymidine incorporation as well as α-methyl glucose and AIB transport activities were mitigated by inhibitors of PKC. Conclusion: Chronic hypoxia induces both proliferation and dedifferentiation of LLC-PK1 cells mediated, in part, by the sustained activation of PKC.

Abstract:Objective: To investigate the effect of reserpine and verapamil on Neisseria gonorrhoeae′s multiple antibiotic resistance. Methods: Rapid quantitative method of minimum inhibitory concentration(MIC) was used for susceptibility testing on 30 strains of N. gonorrhoeae, including standard strain of WHO, A,B,C,D,E strains and strains isolated from outpatients. The influences of reserpine and verapamil on N. gonorrhoeae′s multidrug resistance were tested after they were added to 6 antimicrobials, respectively. Results: A specific concentration of reserpine could reduce MIC of antimicrobials on N.gonorrhoeae strains by 86%,while verapamil by 81%. Conclusion: Reserpine and verapamil have reverse effect on N. gonorrhoeae′s multiple antibiotic resistance, both of them can reduce the antimicrobials′ MIC to N. gonorrhoeae.

Abstract:Objective:To evaluate the protective effects of MgSO4　on neurofunction after cerebral ischemia reperfusion injury. Methods:Twenty-two SD rats were divided into control group, ischemic group and MgSO4 group by means of Pulsinelli and Brierley′s animal model. Meanwhile changes of neurobehavior and pathological changes were observed. Results:Compared with the control and ischemic group, MgSO4 could ameliorate the behavior deficits and improve pathological changes without affecting blood pressure. Conclusion: The data show that MgSO4 may markedly reduce the pathological changes of postischemic brain structures and improve the recovery of neurofunction.

Abstract:Objective: To determine whether there are some differences between GABAA and GABAB receptors in the regulation of cardiovascular activity in the nucleus tractus solitarii(NTS). Methods: Techniques of microinjection into NTS and lesion of NTS as well as electric stimulation of hypothalamic defense area(HDA) were used, and mean arterial pressure(MAP) and heart rate were measured.Results: GABA uptake inhibitor nipecotic acid-induced increase of MAP was blocked by GABAA receptor antagonist bicuculline, but not by GABAB antagonist CGP 35348; glutamate could only partially reverse muscimol-induced elevation of BP while showed no obvious effect on GABAB receptor excitant-induced elevation of BP; and microinjection of bicuculline into NTS could block stimulation of HDA-induced pressor response, but CGP 35348 could not. Conclusion: The differences between GABAA and GABAB receptors in the regulation of cardiovascular activity in NTS may be due to the presynaptic distribution of GABAB receptors and postsynaptic exsistence of GABAA receptors in NTS.

Abstract:Objective:To evaluate the inhibitory effects of norcantharidine(NCTD) on ovarian carcinoma 3AO and AO cells cultured in vitro and to reveal its mechanism.Methods:Using cell culture technique,the inhibitory effects of NCTD on 3AO and AO cells were measured by way of crystal violet dyeing.The alterations of cell cycle induced by NCTD were analyzed with flow cytometry.Results:3AO and AO cells exposed to NCTD 1 μg/ml were inhibited slightly.The IC50 values of NCTD for them were 20 μg/ml and 10 μg/ml respectively.The growth of 3AO and AO cells was inhibited after being cultured with NCTD for 6 h. The inhibitory rates were increased as time of culture prolonged.The percent age of 3AO and AO cells in G2+M phase were increased obviously and the percentage of cells in S phase were decreased accordingly after treated with NCTD. Conclusion: NCTD has a significant inhibition of ovarian carcinoma 3AO and AO cells,which is in a dose- and time-dependent manner. The mechanism of NCTD-induced antiproliferative effect appears to involve blockade of tumor cells in the G2+M phase of the cell cycle.

Abstract:Objective: To study the effects of Qiling capsule(QLC) on natural killer cell (NK) activity in mice. Methods:The cytotoxicity was detected by 3H-TdR release assay and levels of interferon (IFN) in serum by cellular protected method. Results: QLC had the ability to improve the cytotoxicity which was either normal or inhibited by cyclophosphamide(CY)(P<0.05) in vitro. Moreover,in vivo,QLC could increase the NK activity (P<0.05) and raise the serum levels of IFN(P<0.01)in mice pretreated by CY,but no effect on normal mice. Conclusion: The results inferred that the QLC protection of NK against CY inhibition might be related to enhancement of IFN levels.

Abstract:Objective: To investigate the anti-gastric cancer (GC) effect of LAK cells induced by TNFα with IL-2 and the influences of pretreatment with TNFα on the susceptibility of GC cells to LAK. Methods: Induced by TNFα combined with IL-2 at different concentrations, blocked by TNFα McAb, and after pretreating the GC cells with TNFα, LAK cytotoxicity to GC cells was detected by LDH colometric assay. Results: At low concentrations of IL-2, TNFα increased the cytotoxicity of LAK and the concentration of IL-2 necessary to induce the cytotoxicity to the same extent decreased 10 folds. Pretreatment of GC cells with TNFα made the cytotoxicity of LAK to the GC increase by 9%. Conclusion: TNFα increased the cytotoxicity of LAK induced by IL-2 to GC cells, and the GC cells pretreated with TNFα were more susceptible to LAK.

Abstract:Objective: To investigate the effect of NG-nitro-L-arginine (NO2Arg) on psychical dependence induced by opiates on the basis of improved conditioned place preference model in mice. Methods:The time in biased side was recorded before and after 4 mg/kg NO2Arg administration in morphine and dihydroetorphine psychically dependent mice. Results: 4 mg/kg NO2Arg could significantly increase the time in biased side in morphine and dihydroetorphine psychically dependent mice. The time in biased side was increased from (6.09±2.02), (7.98±0.72) min to (9.31±1.10), (9.48±1.21) min (P<0.01). Conclusion: NO2Arg could reverse the psychical dependence induced by opiates, suggesting that NO plays an important role in the development of opiate psychical dependence.

Abstract:Objective: To observe the effects of stilamin on portal hypertensive gastropathy (PHG) in rats. Methods: Two weeks after portal vein ligation, the changes of hemodynamics, GMBF, PVP , and submucosal blood vessels were observed;plasma glucagon, plasma and gastric mucosal 6-keto-PGF1α were tested after intravenous stilamin, propranolol and saline. Results: GMBF and PVP were reduced whereas submucosal blood vessels were not changed after intravenous stilamin and propranolol.Stilamin decreased plasma glucagon,hindered gastric acid and pepsin secretion, had no effect on MAP and heart rate of PHG rats as compared with control group of PHG, whereas propranolol had no such effects, but it might decrease MAP and heart rate. There were no differences in plasma and gastric mucosal 6-keto-PGF1α levels between all PHG groups. Conclusion: Stilamin reduced GMBF of rats with PHG by hindering glucagon, and had no effect on hemodynamics.

Abstract:Objective: To study the differentiation between leiomyoma (LA) and leiomyosarcoma (LS). Methods: One hundred and ninety cases of LA and LS in gastrointestinal tract were investigated by gross, HE staining, special staining, immunohistochemical staining and electric microscopy. Results: The average value of the diameter of LA was 3.3 cm. Tumor surfaces were smooth. Cut section showed hard texture. Interwall and intracavity type were most commonly found.Density of tumor cells reached 1-2 grade. Tumor cells displayed well-differentiation.Nuclei of tumor cells appeared usually to be thin-stick shape or plump-shape. Mean count of mitotic figure was 0.47/25 HPF. Mitotic activity was not observed in 75%of cases. The average value of the diameter of LS was 8.6 cm. The texture was flesh. The dumbbell-shape was commonly seen. Density of tumor cells reached 2-3 grade. Tumor cells were various in shape, and they displayed less-differentiation. Nuclei of tumor cells were irregular in shape or present with giant nuclei or multinuclei. Mean counting of mitotic figure was 20/25 HPF. Specific staining, immunohistochemical staining and electric microscopy were mainly used for determining the origin of tumor cells.Conclusion: Nine items of diagnostic criteria of LA and LS were proposed, in which mitotic figure counting was the basis of diagnosis of LA and the basis of differentiation of low-degree, moderate-degree and high-degree malignant LS. Immunohistochemical staining and electric microscopy are helpful for determining the origin of smooth muscle tumor.

Abstract:Objective: To study the biocompatibility of Ti-75 (TiAlMoZr). Methods: Six adult mongrel dogs received Ti-75 bars and pure titanium bars surgically in their femurs .The dogs were sacrificed at 3,6 and 12 months post-implantation, respectively. On 3rd and 14th day before their sacrifice, intramuscular injection of tetracycline had been given. The specimens that had been canceled with the implants were cut and sectioned. The bone-implant interfaces were observed under a fluorescence microscope and the photographs were taken. The fluorescence labelled areas were evaluated quantitatively by the computer image analysis system. Results: The percentage of labelled area had no difference between Ti-75 and pure titanium. There was also no difference in the three periods when same material was used. Conclusion: The biocompatibility of Ti-75 and pure titanium is similar.

Abstract:Objective: To compare the outcome of patella fractures fixed by biodegradable tension band fixation(BTB) with Biofix rods and Biopoly ligaments and metallic tension band (MTB)with Kirschner wire and metallic cerclage wire in a randomized study. Methods:Thirty-eight patients(18 with BTB and 20 with MTB) were treated in this group. The follow-up time was 24 (14-32) months. The results were evaluated by radiography and recovery of joint function. Results: The fractures healed in all patients after a mean time of 8 weeks. In BTB group, the clinical outcome was good in 13 patients, fair in 4, and poor in 1. In MTB group, the corresponding figures were 15, 3 and 2. There was no clinical differences between the two methods. Conclusion: Patella fractures can be treated successfully by using biodegradable tension band fixation with no need for a second operation to remove the implants.