BACKGROUND:Few reports are found about the effect of ovariectomized rats' bone histomorphometry parameters using non-destructive dynamic loading system.OBJECTIVE:To observe the effect of different loads situations on the bone histomorphometry parameters in ovariectomized rats.DESIGN,TIME AND SETTING:A controlled observation on the bone histomorphometry was performed in the Biomedical Engineering Laboratory of Jilin University from April 2007 to August 2007.MATERIALS:A total of 35 9-month-old Sprague-Dawley female rats were randomly divided into five groups,including sham operation group,ovariectomized (OVX) control group,OVX loading 1 N group,OVX loading 2 N group,OVX loading 4 N group.There were 7 rats in each group.METHODS:Rats in OVX control group and castration load group were processed into bilateral OVX on the back. The sham operation group only underwent the excision of fat tissues on back,and then sutured. After castration for 1 week,rats were loaded with non-destructive dynamic loading system in the two sides of the tibia,15 minutes a day. The mechanical loads would continue for 4 weeks and the loads were 1N,2N and 4N.MAIN OUTCOME MEASURES:Changes of proximal tibia bone histomorphometry parameters.RESULTS:The area,number and thickness of trabecular bone in OVX loading group were all higher than OVX control group (P＜0.05,P＜0.01).The trabecular bone area and thickness in OVX 4 N and OVX 2 N groups were significantly higher than OVX control group (P ＜ 0.001).There was a downward trend of trabecular separation in OVX 4 N group compared with OVX control group (P ＜ 0.05). With the increasing loads,there was an increasing trend of the area,number and thickness of trabecular bone,which were close to sham-operated group. The trabecular separation was declined. Single fluorescent labeled surface and double fluorescent labeled surfaces in sham operated group were all lower than that in OVX control group. With the increase in loads,the single fluorescent labeled surface,double fluorescent labeled surface,interlabeled width and mineral apposition rate had been shown to increase. The OVX 2 N and OVX 4 N groups exhibited a remarkably higher level of mineral apposition rate than OVX control group (P ＜ 0.05).CONCLUSION:With the increase in load at the range of 1-4 N,all parameters of bone histomorphometry improve in the OVX rats,the bone microstrcture is greatly ameliorated,bone mass loss is reduced and the process of osteoporosis is delayed.

BACKGROUND: Nano-hydroxyapatite (Nano-HA) has been shown to be a good choice of bone repair material owing to its salts composition consistent with natural bone and its scaffold structure homothetic to natural bone structure. OBJECTIVE: To investigate the feasibility of nano-HA in repair of jaw bone defect in rabbits. DESIGN, TIME AND SETTING: Materials-based animal experimental observation was performed at Laboratory Animal Center of Jiamusi University and Beijing Ji Shui Tan Hospital between January 2003 and June 2005. MATERIALS: Neutralization reaction of calcium biphosphate and calcium hydroxide was used to construct the system. The reactants were managed to be cotloidal by reaction control and using appropriate nucleating agents. Acerata HA crystal obtained under different conditions was sintered to obtain the nano-HA granule with a diameter of 1-56 nm. METHODS: Twenty-four healthy Chinese Harbin rabbits were randomly and evenly divided into an experimental group and a control group. After anesthesia, a penetrating 1.5 cm x1.5 cm defect was made with the GX micro-drill at the mandibular edge in each rabbit. Nano-HA was implanted in defects of the experimental group, and common HA was used in the control group. Antibiotics were used for 5 days afterwards. MAIN OUTCOME MEASURES: Change in bone mineral density after implantation of nano-HA. RESULTS: After repair of bone defect, the bone mineral density in the expedmental group increased gradually to a normal level and tended to be stable; whereas it was gradually decreased in the control group. There was significant difference between the experimental and control groups (P < 0.01). CONCLUSION: Nano-HA can promote new bone maturation, and this material produces favorable results in repair of bone defects.

Objective Classification of non-small cell lung lymph (NSCLC) node (N) is one of the key factors influencing treatment, however, the cilinical noninvasive and invasive approaches to N classification have their limitations.This study aimed to investigate the risk factors of lymph node metastasis of peripheral lungadenocarcinoma by using CT and PET / CT scans.Methods Retrospective analysis had been done on a total of 248 patients who underwent surgical resection from February 2010 to November 2015 in our hospital.All of them underwent chest CT and 80 patients underwent PET/CT examination.Univariate analysis was applied in the relation of lymph node metastasis to gender, age, smoking situation, CEA, SUV, cancer size, pathological variants, and the degree of differentiation.Multivariable logistic regression analysiss were performed in the prediction of risk factors for lymph node metastasis.ResultsSeventy-four patients (29.8%) had regional lymph node metastases.Univariate analysis showed that lymph node metastasis was related to the serum CEA level, degree of differentiation, SUVmax, tumor size, lobulation/spiculation, pleural retraction, mediastinal or hilar lymphadenopathy (P<0.05).In the multivariable analysis of risk factors, including serum CEA, SUVmax and CT features, for predicting lymph node metastasis, the most important and significantly independent risk factors identified were SUVmax, CEA level, mediastinal or hilar lymphadenopathy, cavitation/bubble-likelucency and pleural retraction (P<0.05).Conclusion The lymph node metastasis is associated with SUVmax of primary tumor, serum CEA level, mediastinal or hilar lymphadenopathy, cavitation/bubble-likelucency and pleural retraction.The combination of radiographic features and serum CEA can help to predict more accurately the risk of lymph node metastasis in patients with peripheral lung adenocarcinoma.

BACKGROUND: Spinal cord injury occurs frequently and its consequence is very severe. There is no effective method to rebuild the function of demylinated nerves. Transplantation of a kind of special glial cells in olfactory system of mammal attracts more attention.OBJECTIVE: To observe whether combined transplantation of human fetal olfactory ensheathing cells (human OECs) and rat embryonic spinal cord tissues (rat ECS) possesses synergistic effect in promoting axonal regeneration in the rats following spinal cord transection.transection.DESIGN: Open experiment.SETTING: Cell Room, Third People's Hospital of Wuxi City; Department of Neurology, First Hospital Affiliated to Soochow University; Department of Neurosurgery, Zhongda Hospital Affiliated to Central South University MATERIALS: This experiment was carried out in the cell laboratory of the Third People's Hospital of Wuxi from September 2002 to October 2004. ① Totally 36 adult female SD rats, of clean grade, were selected and randomly divided into 4 groups: human OECs group (n=10), rat ECS group (n=10), combined transplantation group (n=10) and sham-operation group (n=6). ② Fresh 12-week aborted human embryo was used for culture and purification of human OECs (Informed consent was obtained from the parturient). ③One SD rat at embryonic 14 days underwent caesarean operation, and fetal rat and fetal membrane were taken out together and used for preparing new embryonic spinal cord.METHODS: ①Rats of 4 groups were all created into hemisection cavity models. Gelatin sponge and complete culture medium of 8 μL were packed into the injured cavity of rats in the model group, and the same culture medium of 2 μL was injected at 1 mm above or below injure; Human OECs suspension of 8 μL was added to gelatin sponge in human fetal Human OECs group, and human OECs suspension of 2 μL was injected at 1 mm above and below injure; rat embryonic spinal cord tissue of rat ECS group was chipped into pieces, which were packed into the cavity,and gelatin sponge was spread on the injury part. Embryonic spinal cord with the same size was packed into the cavity of combined transplantation group, then 8 μL human OECs suspension was injected into cavity with micro sample injector, and gelatin sponge was spread on the injury part, and then cellular suspension of 2 μL was injected at 1 mm above and below the cavity, and muscular layer skin was sutured layer by layer. ②The rats of each group were performed ethological evaluation periodically. Combined with pathological observation, effect of human OECS and rat ECS on neuronal survival and regeneration was evaluated by performing horseradish peroxidase-tetramethyl benzidine tracer technique.MAIN OUTCOME MEASURES: ①In vitro culture and purification of human fetal human OECs. ② In vitro immunocytochemistrical analysis. ③BBB scoring of motor function of hindlimb of rats. ④ Immunohistochemical detection of implants and injured spinal cord repair⑤ Quantitative analysis on labeled neurons at the cortex and mesencephalic red nucleus ineach group with horseradish peroxidase-tetramethyl benzidine tracer technique.RESULTS: ① Most of human fetal OSCs presented double-polar spindle.Five to seven days after culture, OSCs weaved into net and a lot of mitosis phases were found. The cellular purity was 85%. ② The rate of P75 positive cells was (83±7)%. Glial fibrillary acidic protein was found in about (81±6)% of cells and Vimentin in (91±9)% of cells and the rate of Nestin positive was (77±5)%. ③Three to five days after operation, affected limb of rats of sham-operation group began to contract, the activity of hindlimb of intact side was limited a little. Fewer obvious contraction symptoms were found in the other 3 groups. From 2 weeks after operation,behavioral function recovered significantly fast in each group. BBB scores of combined transplanted group were significantly high than those of human OECs group, rat ECS group and sham-operation group [(6.2±1.13) vs.(5.0±1.15)vs.(3.9±0.88)vs.(3.3±1.03)scores,P ＜ 0.05]. ④In bipolar or multipolar cells, in which basic protein(+)granules were found, P75 and glial fibrillary acidic protein positive were found at the implanted part in the range of 2.0 to 5.0 mm of transplanted region in the human OECs group and combined transplantation group. A great many of small MAP2 positive neurons were found in the spinal defected focus in the rat ECS group and combined transplantation group. Nerve plexus positive fibers were observed in spinal defected region of human OECs group, rat ECS group and combined transplantation group to different extents, especially significantly in the combined transplantation group, but they were not found in the model group. ⑤ Horse radish peroxidase labeling was hardly found in neurons at the injured side of sham-operation group, while the number of labeled neurons at the cortex and nesencephalic red nucleus was significantly higher in the combined transplantation group than in the human OECs group and rat ECS group (P ＜ 0.05).CONCLUSION: Combined transplantation of OECs and ESC can obviously protect injured spinal cord, promote host spinal axonal regeneration and play s a complementary and synergetic effect in speeding up the functional recovery of rats.

Objective To establish a human keratinocyte cell line (HaCaT) stably expressing HPV16E7 protein. Methods HPV16E7 gene was amplified from CaSki cells using PCR and inserted into the eukaryotic expression plasmid pcDNA3.1. Then, the recombinant expression plasmid pcDNA3.1-HPV16E7 was transfected into HaCaT cells followed by G418 selection and identification by RT-PCR and Western blot. Results The recombinant eukaryotic expression plasmid pcDNA3.1-HPV16E7 was successfully identified by restriction enzyme digestion pattern and sequence analysis. Agarose gel electrophoresis of RT-PCR products detected the 297-bp fragment of HPV16E7 cDNA, and Western blot confirmed the stable expression of HPV16E7 protein. Conclusion A human keratinocyte cell line (HaCaT) stably expressing HPV16E7 protein is successfully established.

Objective To establish a model for cutaneous squamous cell carcinoma by irradiation of SKH-1 hairless mice with solar-simulated ultraviolet (solar UV), and to explore the biological characteristics of the model. Methods A total of 91 SKH-1 hairless mice were randomly divided into seven experimental groups (n = 10) and seven control groups (n = 3). The mice in experimental groups were irradiated with minimal erythema dose of solar UV 4 times per week for various durations (4, 8, 12, 16, 20, 24, 28 weeks), while the control mice received no irradiation. The general status and skin appearance of mice were observed during the treatment process. Mice were killed immediately after the last irradiation at different time points and pathological examination was carried out to observe the histological changes of skin lesions. Results Papules measuring equal to or more than 1 mm in diameter began to develop in some mice in experimental group 10 weeks after the first irradiation; tumors began to appear in 39.3% (11/28) of the remaining mice in experimental group on week 20, and in 100% (10/10) of the remaining mice on week 28. The cumulative dose approximated to 26.99 J/cm2 for UVB and 242.91 J/cm2 for UVA after 28-week irradiation. No tumor was observed in the control mice. Pathological examination revealed characteristic changes of squamous cell carcinoma in 30% of the mice on week 12, 33.3% on week 16, 60% on week 20, 87% on week 24, and 100% on week 28. Conclusions Ultraviolet could induce the hyperplasia of skin in SKH-1 hairless mice, and even cause the development of cutaneous squamous cell carcinoma after prolonged irradiation.

Objective To determinate the risk factors of gout in patients with hyperuricemia.Methods Patients detected with hyperuricemia both in epidemiological survey of Shandong coastal areas in 2004 and in health examination of our hospital were followed up for three years to observe the incidence of gout, relationship of diet and gout, and changes of biochemical indicators.Results During 3 years, 102 patients (19%) out of 536 patients with hyperuricemia developed gout. Age(OR=1.046, P＜0.05), serum uric acid(OR=1.021, P＜0.05), fasting plasma glucose(OR=1.021, P＜0.05), triglyceride(OR=1.008, P＜0.05), tony crab intake ( OR=5.992, P＜0.05),and beer intake(OR=1.012, P＜0.05) were the risk factors of gout attack in patients with hyperuricemia.Conclusions Excess intake of tony crab and beer resulting in fluctuation of serum uric acid is the main risk factor of gout in patients with hyperuricemia. Correcting metabolic disorder of glucose and lipid, reducing the intake of high-purine food, and controlling the level of serum uric acid are the measures to reduce gout attack.

Objective To investigate the effect of safranal on neurologic functions and histopathologic changes after spinal cord injury (SCI)and the related molecular mechanisms.Methods We randomly assigned 36 rats into six groups:control group,injury group and four treatment groups (namely,A,B,C,and D).The Basso Beattie Bresnahan locomotor rating scale (BBB)and HE staining were applied to evaluate the neuroprotective effect and determine the most effective dosage.Another 60 rats were randomly and evenly assigned to three groups:control group,injury group and treatment group.Nissl staining,TUNEL staining and electron microscopy were used to analyze histopathological changes;RT-PCR,immunohistopathological staining,ELISA,and Western blot were used to detect the expressions of apoptosis-related proteins (Bax and Bcl-2 ),inflammation-related factors (IL-1β, IL-10,TNF-αand P38MAPK),and edema-related factor (APQ-4).Results The optimal dosage for safranal was 100 mg/kg.Neurocyte structure was found more distinct in treatment group than in injury group.In addition,we detected a smaller number of apoptotic neurocyte (26.37±1.54 vs.35.94±1.62,P=0.000),decreased Bax (P=0.000)and APQ-4[(359.55±16.12)% vs.(124.53±20.35)%,P=0.000]expressions,increased Bcl-2 (P=0.036)expression,and obviously lowered P38MAPK [(300.30±33.26)% vs.(132.54±10.21)%,P=0.000]expression. Conclusion Safranal exerts its neuroprotective function through anti-apoptosis,anti-inflammation and anti-edema in the rat model of spinal cord injury.

Objective:To observe the effects of acupuncture at Houxi(SI3)and Huantiao(GB30)on the expression levels of nuclear factor kappa B(NF-κB),inducible nitric oxide synthase(iNOS),and nitric oxide(NO)of NF-κB inflammatory signaling pathway in L5 spinal nerve root of lumbar disc herniation(LDH)model rats and explore the mechanism of acupuncture in LDH treatment.Methods:Forty specific-pathogen-free healthy male Sprague-Dawley rats were randomly divided into a sham operation group,a model group,acupuncture group 1,and acupuncture group 2,with 10 rats in each group.The non-compression nucleus protrusion model was made by puncturing L4-L5 spinous process space and injecting autologous nucleus suspension.Acupuncture at bilateral Shenshu(BL23),Dachangshu(BL25),and Weizhong(BL40)was carried out in acupuncture group 1,and acupuncture at bilateral Houxi(SI3)and Huantiao(GB30)in acupuncture group 2.All rats were treated with balanced reinforcing and reducing needling manipulations,and the needles were retained for 30 min/time with one episode of needling manipulation every 10 min,once a day,14 times in total.The threshold value of paw withdrawal pain was measured by a thermal stimulation pain instrument;the serum NF-κB,iNOS,and NO levels were measured by enzyme-linked immunosorbent assay.The pathomorphological changes of spinal nerve roots were observed by hematoxylin-eosin(HE)staining;quantitative reverse transcription polymerase chain reaction was used to detect iNOS mRNA expression in spinal nerve roots;the NF-κB and iNOS protein expression in spinal nerve roots was detected by the immunofluorescence method.Results:Compared with the sham operation group,the threshold of paw withdrawal pain in the model group was decreased,and the expression levels of serum NF-κB,iNOS,and NO were increased;HE staining showed many degenerated and dissolved Schwann cells in spinal nerve roots with vacuolar changes;meanwhile,the expression levels of NF-κB and iNOS proteins,and the iNOS mRNA in spinal nerve roots were increased.Compared with the model group,the paw withdrawal pain thresholds in acupuncture group 1 and acupuncture group 2 were increased,and the increase in acupuncture group 2 was greater(P<0.05);the expression levels of serum NF-κB,iNOS,and NO in acupuncture group 1 and acupuncture group 2 were decreased,especially in acupuncture group 2(P<0.01);the vacuolar changes of spinal nerve roots,and the degeneration and lysis of Schwann cells in acupuncture group 1 and acupuncture group 2 were decreased,which were more obvious in acupuncture group 2;the NF-κB and iNOS protein expression and the iNOS mRNA expression levels in spinal nerve roots of acupuncture group 1 and acupuncture group 2 were decreased,especially in acupuncture group 2(P<0.01).Conclusion:Acupuncture at Houxi(SI3)and Huantiao(GB30)can improve the morphology of spinal nerve roots,inhibit the NF-κB and iNOS protein expression levels in spinal nerve roots and the serum NO level,and relieve the pain caused by inflammation of spinal nerve roots,which may be one of the mechanisms of acupuncture in LDH treatment.

BACKGROUND: To repair bone defect, histocompatibility, growing characteristics, biodegradation and repairing mechanism of nanometer need to be further studied in clinic. OBJECTIVE: To observe the growing characteristics and histocompatibility of nano-hydroxyapatite (Nano-HA) for repairing jaw defect of rabbits.DESIGN: Randomized grouping animal study.SETTING: Beijing Jishuitan Hospital and Stomatology College of Jiamusi University.MATERIALS: A total of 24 New Zealand rabbits, either gender, weighing 2.5-3.5 kg, were provided by Animal Experimental Center of Jiamusi University. The animal experiment had got confirmed consent from local ethic committee. Nano-HA was provided by Material Engineering College of Jiamusi University and dealt with routine hyperthermia/hypertension sterilization. In addition, hydroxyapatite was provided by Wuhan Industry University, and the diameter was 1.0-2.0 μm.METHODS: The experiment was carried out in the Experimental Animal Center of Jiamusi University from November 2001 to May 2006. All rabbits were randomly divided into experimental group and control group with 12 in each group. Bone defect in the diameter of 1.0 cm was produced on body of mandible. Nano-HA was used to repair the bone defect of rabbits in the experimental group, while hydroxyapatite was used to repair the bone defect of rabbits in the control group. At 1, 4, 8 and 12 weeks after operation, all rabbits were sacrificed. In addition, medical imaging analysis system was used to analyze generative quantity of tissue in the two groups; meanwhile, histological quality and quantity were also analyzed so as to observe histocompatibility and newborn osteogenesis. MAIN OUTCOME MEASURES: Histocompatibility and newborn osteogenesis.RESULTS: With the time passing by, the amount of repairing materials was decreased because of the combination with newborn tissue into bone in bone defect-repaired region in the experimental group. When it was closed to normal bone, the amount was stable. However, bony callus was not able to grow in materials in the control group. Results of correlation analysis demonstrated that materials were negatively straight-line correlation with newborn bone (r = -0.912 0, P < 0.01). During the repairing procedure of bone defect, newborn bone was closely correlative with Nano-HA; while, with the increase of newborn bone, the amount of repairing materials was decreased because of the combination with newborn tissue into bone.CONCLUSION: Nano-HA can combine with newborn bone tissue so as to rapidly generate bone, while it has an excellent biocompatibility.