BACKGROUND:Tissue engineering requires a lot of seed cells. Osteoblasts have become important seed cells in bone tissue engineering. However, it is difficult to culture the osteoblasts, and cellnumber, purity, proliferation and differentiation activity are different obtained by different culture methods. OBJECTIVE:To identify and compare three common primary osteoblat culture methods, and to explore a method for the primary culture of osteoblasts which is easy to operate, economical and effective, in order to provide basis for the further experimental research. METHODS:Calvarias were dissected from newborn Sprague Dawley rats in 72 hours, and osteoblasts were isolated with col agenase digestion method, sequential digestion method and bone tissue method respectively. The morphological observation and cytochemical staining were performed, the growth curve of the cells was drawn with cellCounting Kit-8 method, and the rate of living osteobalsts was counted with trypan blue staining. RESULTS AND CONCLUSION:The proliferation of the insolated and cultured osteoblasts was wel with typical characteristics of osteoblasts, cytochemical staining results were positive. Compared with the sequential col agenase digestion method, the col agenase digestion method presented higher production of osteoblasts and higher cellsurvival rate (P<0.05), and the col agenase digestion method was easier than the sequential col agenase digestion method and cost less than sequential col agenase digestion method. Bone tissue method was the easiest method with less damage to cells, but bone tissue method presented lower production of osteoblasts and cost much more time, which cannot be used in large-scale osteoblast culture. The col agenase digestion method is a simple, efficient and ideal method for isolation and culture of primary osteoblasts.

In order to improve the efficacy of vibration expectoration and prevent secondary injury in patients with chest trauma,the body surface marking method was used in vibration expectoration.The exact injured locations and areas were identified via CT ot X-ray results.Then,the restricted and focused areas of expectoration were marked with red and green colors,which could guide the machines to avoid the fracture sites,reduce pain,prevent secondary injury and administrate effective expectoration in the areas of sputum and atelectasis.However,the dynamic CT or X-ray results must be used for reference to ensure the accuracy of the body surface marking.

Objective To investigate the role of nerve growth factor (NGF) in the pathogenesis of diabetic neuropathy (DNP) and the correlation of NGF level with,sensory nerve conduction velocity (SNCV) and duration of DNP.MethodsThe 41 patients with DNP,35 diabetics without DNP and 33 healthy controls were enrolled.And the serum level of NGF was measured with ELISA,the SNCV of left medial nerve,ulnar nerve,peroneal nerve and tibial nerve were measured too. Then the correlation analysis was completed. ResultsThe serum level of NGF was significantly lower in DNP patients [(665.18±188.32) ng/L] than in healthy controls [(976.44±159.07)ng/L] and in diabetics without DNP [(943.32±167.33) ng/L,F=9.316,P<0.001].The NGF level of DNP patients was positively correlated with SNCV of left medial nerve (r=0.810,P<0.001 ),left peroneal nerve (r=0.760,P<0.001) and duration of DNP (r=0.542,P<0.001).Conclusions The serum level of NGF is lower in DNP patients than in healthy controls.The decrease of the production of NGF may play a role in the pathogenesis of DNP.

Objective To investigate the effect of perinatal bisphenol A BPA exposure on brain development of F1 male offspring. Methods Pregnant SD rats were given BPA at 2 20 and 100 mg/kg body weight per day respectively from eleventh day of gestation to the whole lactation by gavage until their pups were weaned on postnatal day 21  the control group had no BPA exposure. Every six F1 male pups from each of the four groups were killed at differential time points on postnatal day 1510152130 and 45 respectively. Histopathological examination by HE stain was done on the brains. Results The results showed no abnormal change was found on postnatal day 1-10. Three dosage groups showed abnormal change of different degree on 15th 21th 30th postnatal day the mainly abnormal change was karyopyknosis of pyramidal cell in CA3 of hippocampus and cortical neuron in cerebral cortex. The cell numbers of pyramidal cell in CA3 of hippocampus and cerebral cortex were decreased on 45th postnatal day. Conclusion Perinatal BPA exposure may have an adverse effect on the brain developmnent of F1 male offspring.

Objective To investigate the effects of electromagnetic irradiation on the activity of protein kinase C(PKC)in the cerebellum of rabbit.Methods Thirty rabbits were randomly divided into control and electromagnetic irradi.ation group (including 0 hour,3 hours,12 hours,24 hours and 72 hours subgroups after irradiation),rabbits in the electromagnetic irradiation group were exposed to 65 mW/cm~2 S frequency range electromagnetic irradiation for 30 minutes,the rectal temperature was taken immediately after irradiation and the specific absorption rate(SAR)values were calculated,the activity of PKC the cerebellum was determined with improved TaKai method.Results The rectal temperature of rabbits increased 2.35 ℃ after irradiation,SAR value was 4.54 kcal/kg.PKC activity decreased significantly after irradiation.Conclusion After 30 min of electromagnetic irradiation at 65 mW/cm~2,obvious heating effect will be detected in the rabbit and PKC activity in the cerebellum decreased significantly.

Objective PB840 ventilator to explore non-invasive ventilation (NIV) in the ICU of the application,efficacy and safety.Methods Application PB840 Ventilator-Non-invasive mechanical ventilation in the treatment of the seven cases of different causes of respiratory failure patients,non-invasive ventilation in the treatment of 4 h,the blood gas (PaO2,PaCO2 and pH value) analysis and analysis of clinical symptoms to improve the situation.Results Patient before and after treatment of PaO2,PaCO2 and pH compared to the value there is significant difference.Target to improve the blood after treatment significantly (P<0.05),reduce or alleviate the symptoms of sick;cyanosis improved.However,leakage is still a major disadvantage,some patients can not tolerate when there is need to immediately replace invasive mechanical ventilation.Conclusion PB840 ventilator mode non-invasive ventilation can improve the patient′s respiratory failure and clinical symptoms of blood-gas target,but most critically ill patients should be carefully applied.

Objective To investigate the clinical effect of shuxuening treatment on organophosphorus pesti-cide poisoning with toxic myocarditis.Methods 60 patients with organophosphorus toxic myocarditis were selected in our hospital as the research subjects,and 60 cases were divided into two groups:observation group(n =30) and control group(n =30).Control group was given conventional treatment,treatment group was given shuxuening injec-tion 14 days on the basis of conventional treatment.After treatment,creatine kinase isoenzyme (CKMB),troponin (TNI),interleukin 6 (IL -6) and cholinesterase(ChE) were compared,and the changes of clinical symptoms were observed at the same time.Results There was no significant difference between the observation group and the control group(χ2 =0.630,P =0.730).The TNI,IL -6,CKMB could reflect the severity of myocardial injury in patients with different degrees of organic phosphorus poisoning,TNI,CK -MB,IL -6 increased with the degree of poisoning,the differences were statistically significant(F =11.863,4.512,3.774;P =0.000,0.015,0.029).After treatment for 4, 9,14 days,TnI,CK,CK -MB,levels of IL -6 in the two groups were recovered,but the recovery levels of TnI,CK -MB and IL -6 of the observation group significantly better than the control group,the differences were statistically sig-nificant(Fourth days,t =8.125,5.128,10.461;P =0.000,0.001,0.000;Ninth days,t =5.464,4.674,9.510;P =0.001,0.002,0.000,t =6.162,8.248,5.523;P =0.000,0.000,0.001).Conclusion Conventional treatment combined with shuxuening in the treatment of organophosphorus pesticide poisoning with toxic myocarditis has better therapeutic effect,it is worthy of promotion.

Objective To investigate the clinical effect of super selective embolization combined with opera-tion for treatment of meningiomas.Methods 82 cases treated by operation and pathology in return for meningioma were collected in the study,including 44 cases of preoperative superselective embolization for aneurysms as the treat-ment group,and the remaining 38 cases without preoperative embolization of patients as the control group.The opera-tion time,bleeding amount in the operation and the rate of total tumor removal were recorded and compared between the two groups.Results The average amount of bleeding of treatment group was (330.36 ±95.41)mL.The average amount of bleeding of control group was (460.82 ±114.64)mL.The data of the two groups was statistically significant (t =5.624,P <0.01).,The treatment group bleeding amount was less than that of the control group;,the mean oper-ation time of treatment group was (5.36 ±2.62)h,,the average operation time of control group was (6.63 ±2.25)h.The data of the two groups was statistically significant (t =2.355,P =0.021 ),the operation time of the treatment group was less than that of the control group.The total resection rate was 70.4% in treatment group and 36.8% in the control group,there was statistically significant (P <0.05),and the total resection rate in the treatment group was higher than that in the control group.Conclusion Compared with single operation treatment of meningiomas superse-lective embolization combined with operation for treatment of meningiomas can shorten operation time,reduce intraoperative hemorrhage,improve the rate of total tumor removal.It can reduce the operation risk and improve the treatment effect.

Objective:To investigate the immune mechanism of 5-fluorouracil(5-Fu) and Octerotide(Oct)in the treatment of severe acute pancreatitis(SAP)and it’s kidney injury.Methods:55 male SAP rats induced by retrograde infusion of 5% sodium taurocholate into bilopancreatic duct were randomly divided into 3 groups:Normal saline group(n=17)were treatmented with normal saline infusion,Oct group(n=18):The first dose of Oct was 2?g/100g iv,then 0.2?g/(100g?h)continuous injection,5-Fu group(n=20):The dose of 5-Fu was 2mg/h?12h.Rats from each group were killed at 12h after opertation.The plasma levels of tumor necrosis factor-?(TNF-?),interleukin-1(IL-1)were assessed by ELISA method.Thromboxan B_2(TXB_2)and prostaglandinE_2(PGE_2)were assessed by radioimmunoassay method.The expression of TNF-? mRNA was detected by RT-PCR.The apoptosis rate of rat kidney tissue were detected by TUNEL method.Apoptosis rate of NRK cell treated by serum of different groups were assessed by flow cytometry methods.Results:The plasma level of TNF-?、IL-1 and TXB_2 in 5-Fu and Oct group were much lower than that in normal saline group.The expression of TNF-? mRNA of kidney tissue decreased significantly after the SAP was treated by 5-Fu or octreotide.The apoptosis rate of NRK cell disposed by normal saline group,oct group,5-Fu group and control group rat plasma were (38.67?11.4)%,(20.4?18.4)%,(10.5?11.0)% and (1.7?2.2)% respectively(P

Objective To investigate the possible protective effect of neurotropin on the corresponding spinal cord motoneuron after sciatic nerve injury early in rats.Methods 108 male Sprague Dawley rats were divided randomly into Groups A,B and C with 36 animals in each group,and all animals were cut both side of sciatic nerve,instantly make the operation of end-to-end anastomose of epineurium for abscised sciatic nerve.Group A was control group.Group B was treated with neurotropin and Group C was treated with normal saline after operation.The rats were sacrificed on 1st d,4th d,9th d,14th d,21st d and 28th d after operation.The corresponding spinal cord segments(L4～L6)were gotten to make Bcl-2 and Bax immunohistochemical staining and TUNEL staining.Results The values of Bcl-2/Bax on 9th d,14th d and 21st d after operation of Group B were different from that of Group A and Group C.The number of positive cells stained by TUNEL staining on 9th d was significantly different among Group A,Group B and Group C(v<0.05).The expression of positive TUNEL staining cells reached a peak on 14th d after operation,the number of Group B was less than that of Group A and Group C.Conclusion After instantly make the operation of end-to-end anastomose of epineurium for abscised sciatic nerve,neurotropin can protect the spinal cord motoneurons to avoid apoptosis excessively in some extent.