BACKGROUND: Tetramethylpyrazine has the protective effect against the central nervous system injury. The structural changes in Nissl bodies were regarded as a marker of neuron injury.OBJECTIVE: To investigate the effect of tetramethylpyrazine on the structure and the quantity of Nissl bodies of cerebral neurons in rat with epilepsy.DESIGN: A comparative study.SETTING: It was conducted at the Physiological Department of Medical School of Xianning College.MATERIALS: From September 2004 to March 2005, it was completed at the Anatomy Department of Tongji Medical College, Huazhong University of Science and Technology. Forty healthy SD rats aging 3-4 months, weighing (250±50) g and regardless of their gender, were selected.METHODS: Rats underwent anesthesia and craniotomy. Then their cerebral cortex were exposed for placing BL-410 Experimental System of Biological Function (TME, China) to record the bilateral EEG of the brain and the seizure in rats with penicillin-induced epilepsy group and the 10 mg/kg tetramethylpyrazine group, 20 mg/kg tetramethylpyrazine group and 40 mg/kg tetramethylpyrazine group. In control groups, the brains of rats were taken out at 1 hour after craniotomy. In penicillin-induced epilepsy group, their brains were taken out at 1 hour after penicillin-induced epilepsy. In 10 mg/kg tetramethylpyrazine group, 20 mg/kg tetramethylpyrazine group and 40 mg/kg tetramethylpyrazine group, after stable penicillin-induced epilepsy, tetramethylpyrazine was injected intraperitoneally at a dose of 10 mg/kg, 20 mg/kg and 40 mg/kg, respectively.When the greatest protective effect of tetramethylpyrazine appeared, the rats' brains were taken out. Brain sections were sliced. Nissl bodies were stained by thionine staining. Under light microscope, structures of Nissl bodies were observed and the images of Nissl bodies were quantitatively analyzed by HPIAS-1000 high acuity color pathologic diagram-writing analyzing system. In each group, the average absorbency of 15 fields was regarded as the average absorbency of Nissl bodies in that group.MAIN OUTCOME MEASURES: In all the groups, the structure and the quantity of Nissl bodies of cortical neurons in rats were studied.of the structure of Nissl bodies in external granular layer cells and external pyramidal layer cells. In control group, multi-layer blue-stained and clumplike or granule-like Nissl bodies could be observed. In penicillin-induced epilepsy group, Nissl bodies were completely resolved and disappeared. In 10 mg/kg tetramethylpyrazine group, Nissl bodies were partly or completely resolved. In 20 mg/kg tetramethylpyrazine group, the quantity of Nissl bodies was significantly increased as compared with those in penicillin-induced epilepsy group and the 10 mg/kg tetramethylpyrazine group. In 40 mg/kg tetramethylpyrazine group, the quantity and the structure of of the average absorbency of stained Nissl bodies in external granular layer cells and external pyramidal layer cells in rat cortex among all the groups.In penicillin-induced epilepsy group, the average absorbency were dramatically lower than that in control group (0.033±0.002, 0.756±0.035, t=4.93,P ＜ 0.01). In 20 mg/kg tetramethylpyrazine group and 40 mg/kg tetramethylpyrazine group, the average absorbency were significantly higher than that in penicillin-induced epilepsy group (0.435±0.011, 0.658±0.029, t=2.98,5.32, P ＜ 0.01). In 40 mg/kg tetramethylpyrazine group, the average absorbency were similar to that in control group (t=1.75, P ＞ 0.05).CONCLUSION: Tetramethylpyrazine can significantly elevated the concentration of Nissl bodies of neurons in rats with epilepsy. Changes in the structure and quantity of Nissl bodies of cerebral neurons may be closely associated with seizure, and tetramethylpyrazine can restore the structure and the quantity of Nissl bodies of neurons, regulate their functions and hereby, inhibit seizures.

Objective: To investigate the effect of vitamin B12 on the growth and hemopoiesis of broilers. Methods: The Arbor Acres broilers were fed the experimental diets with different doses of vitamin B12 0.00, 0.008, 0.016, 0.024 mg/kg for 3 weeks. The body weight gain and hematologic indices of the broilers were measured. Results: Vitamin B12 deficiency in the diet could depress the body weight gain and feed conversion ratio of the broilers, decrease the amount of red blood cell and hemoglobin, enlarge the volume of red blood cell, increase the content of each cell hemoglobin, reduce platelet in blood. While 0.008 mg/kg vitamin B12 was added in the experimental diets, the body weight gain and feed conversion ratio of the broilers increased markedly. Furthermore, when 0.016mg/kg vitamin B12 was added to the diet, the number of the red blood cell and the content of hemoglobin of the broilers increased significantly, and the shape and volume of the red blood cell became ordinary. There were significants interrelations between BWG,RBC,Hb,PCV,PLT,MCV,MCH of the broilers and the addition of vitamin B12 in the diet. Conclusion: Vitamin B12 deficiency could result in the megaloblastic animia. 0.02-0.03 mg vitamin B12 per kilo diet was required to maintain the growth and normal hemopoiesis of the broilers.

BACKGROUND:In an additional changing magnetic field, magnetosomes can invade the tumor cells and inhibit their proliferation.OBJECTIVE: To study the effect of targeted delivery of pHSP70-shPLK1/DOX compounds with magnetosome of magnetotactic bacteria on the treatment of osteosarcoma.METHODS: Human osteosarcoma cell U2OS was divided into four groups: group A: magnetosomes of magnetotactic bacteria+pHSP70-shPLK1/DOX complex; group B: magnetosomes of magnetotactic bacteria; group C: magnetosomes of magnetotactic bacteria+pHSP70-shPLK1/DOX complex, combined with magnetic intervention; group D: magnetosomes of magnetotactic bacteria, combined with magnetic intervention. After 12, 24, 48 and 72 hours of culture, the uptake rate of osteosarcoma cells was observed; cell cycle was evaluated by flow cytometry; PLK1 mRNA and protein expression levels were detected by RT-PCR and western blot, respectively; cell proliferation was explored by MTT assay; cell adhesion and invasion were checked by adhesion assay and Transwel chamber assay, respectively; and the apoptosis rate was measured by apoptosis detection Kit.RESULTS AND CONCLUSION: (1) After culture of 12 and 24 hours, the uptake rate was the highest in group D; meanwhile, the uptake rate was the highest in group B after 48 and 72 hours of culture. (2) The ratio of G2/M phase decreased and the ratio of G0/G1 increased gradualy in group A, group C and group D; the G2/M phase ratio of C group was the lowest at each time point, followed by group D, and that of group A was the highest; and the G2/M phase ratio increased gradualy in group B. (3) Folowing 12 and 24 hours of culture, the expression of PLK1 in group B and group D was higher than that in group A and group C (P < 0.05); in addition, the expression of PLK1 in group B was higher than that in the other three groups after 48 and 72 hours of culture (P < 0.05), and the PLK1 level in group D was higher than that in group A and group C (P < 0.05). (4) The proliferation, adhesion and invasion ability in group B were the highest at different time points, but the apoptosis rate was the lowest. Furthermore, the proliferation, adhesion and invasion abilities in group C were the lowest at different time points, and the apoptosis rate was the highest. To conclude, our experimental results show that the targeted delivery of pHSP70-shPLK1/DOX complex by magnetosomes of magnetotactic bacteria can enhance the apoptosis of osteosarcoma cells and inhibit their proliferation and invasion.

Objective To observe the effects of interleukin-1β (IL-1β) and pentylenetetrazol (PTZ) induced acute epilepsy and the dynamic expression of aquaporin-4 (AQP4) in hippocampus. To explore the role of IL-1β in the pathogenesis of epilepsy by regulating AQP4. Methods All rats were randomly divided into control group, IL-1β group, PTZ group, IL-1ra + PTZ group and dexamethasone + PTZ group. Observe the behavior of the rats within 60 minutes after injection and record seizure score in each group. Then immunohistochemistry and RT-qPCR were used to detect the expression of AQP4 at at 6 , 12, 24 and 36 h. Results Almost of rats in IL-1β group and PTZ group showed severe degree seizure. The rats in control group and dexamethasone + PTZ group showed no obvious seizure. The seizure of rats were more remarkable serious in PTZ group than that in the IL-1ra + pentylenetetrazole group (P < 0.05). Immunohistochemistry and RT-qPCR Show: the expression of AQP4 in hippocampus in PTZ group increased gradually after 12 h (P < 0.05), then reached in the peak after 24 h (P < 0.001). The expression of AQP4 in IL-1ra + PTZ group was lower compare with PTZ group in each time (P < 0.05). Although the expression of AQP4 in dexamethasone + PTZ group higher than the control group, it was not significantly different (P < 0.05). Conclusion The proinflammatory cytokine IL-1β break the balance of water in brain and increasing the concentration of extracellular excitatory amino acids or ions by upregulate the expression of AQP4 in order to promote the excitatory of neurons.

Objective Toinvestigatethepossibilityandquantitativerangeofpepsinogen(PG)usedas the screening marker of gastric cancer by detecting serum pepsinogen level in different gastric mucous pathologic status.Method ThelevelofserumpepsinogenⅠ(PGⅠ)andpepsinogenⅡ(PGⅡ)bytimeresolvedfluoro-immunoassay(TRFIA)in 64 chronic atrophic gastritis patients,63 gastric mucous atypical hyperplasia patients, 67 gastric cancer patients and 20 healthy volunteers were defeeted ,and the ratio of PGR(PGⅠ/PGⅡ)was calculated.Then the three kinds of diseases were graded.The data was analyzed between groups and sub-groups.Result ①Compared with normal control group,the median PGⅠvalues were 1 24.01 ,91 .23 and 71 .23 respectively,which were all lower than that of healthy group (1 52.00).There were significant differ-ences(Z=-2.52,P=0.01 7 0;Z=-3.42,P=0.001 4;Z=-3.57,P=0.000 9).The median PGR values were 7.61 ,5.21 and 4.32 respectively,which were also lower than that of healthy group,the differences were significant(Z=-2.98,P=0.002 9;Z=-3.1 7,P=0.000 2;Z=-2.89,P=0.000 1 ).The PGⅡlevel of these diseases were not significantly different with control group.②The serum PGⅠlevel of gastric mucous atypical hyperplasia and gastric cancer were reduced significantly in contrast with atrophic gastritis (Z =-3.42,P =0.001 4;Z=-3.62,P=0.000 9);the levels of PGⅡand PGR were varied without significance (P>0.05 );③The levels of PGⅠamong atrophy gastritis and gastric cancer subgroup have no significant difference(χ2 =2.86,P=0.41 4 3;χ2 =1 .67,P=0.1 36 8).But the level of PGⅠwas significantly different in gastric atypical hyperplasia(χ2 =0.83,P=0.043 0).It decreased in light and medium grade dysplasia and went up in severe grade dysplasia.The levels of PGⅡ and PGR were varied without significance(P>0.05).④The areas under the ROC curves performed by the PGⅠ and PGR from normal control group and atypical hyperplasia group were 0.782 and 0.831 respectively;The sensitivity and specificity of PGⅠ≤72.1 2 μg/L and PGR≤4.32 for gastric dysplasia were 89.48%and 76.31%respectively.Conclusion ①The level of PGⅠand PGR were decreased along with the seriousness of gastric pathological changes and probably regarded as the screening markers of gastric mucous malignant transformation.②Serum pepsinogen level is closely correlated with gastric precancerous lesion,PGI≤72.1 2 μg/L and PGR≤4.32 has better specificity and sensitivity for gastric atypical hyperplasia in this area.

Objective To analyze the role of a key intracellular signaling molecule GSK3β in hepatocyte apoptosis induced by endoplasmic reticulum stress (ERS).Methods Using mouse hepatoma cell lines(Hepa 1) as cell apoptosis model triggered by tunicamycin,an endoplasmic reticulum stress inducer.One hour before Hepa 1 apoptosis induced by tunicamycin,SB216763 specifically inhibited the activity of GSK3β.Living cells/apoptotic cells were detected using acetoxymethyl (AM)/propidium iodide (PI) staining; Furthermore,the measurement of lactate dehydrogenase(LDH) of cell culture supernatant to evaluate the apoptosis.We detect p-GSK3β,GSK3β,the ERS-related protein(GRP78,CHOP and caspase-12) and caspase-3,cleaved caspase-3 protein expression using Western blot.Results Endoplasmic reticulum stress induced by tunicamycin promotes GSK3β activity; Inhibition of GSK3β activity alleviates endoplasmic reticulum stress:the expression of GRP78,CHOP and caspase-12 expression are inhibited.At the same time,GSK3β activity inhibition significantly reduced the endoplasmic reticulum stress-induced apoptosis:compared to cell apoptosis model group,the intervention group of SB216763 showed that the level of LDH decreased significantly,and PI staining of apoptotic cells was also significant reduction.Western blot results showed that the inhibition of GSK 3 β activity reduced reactive cleaved caspase-3 protein.Conclusion GSK3β is an important signaling molecule in the apoptosis pathway induced by endoplasmic reticulum stress ;Endoplasmic reticulum stress promotes hepatocyte apoptosis by mediating GSK3β.

Objective To investigate the dynamic expression of the drug resistance protein P-glycoprotein (P-gp) within 72 hours in the pentylenetetrazol (PTZ)-induced status epilepticus (SE) model, and to identify the optimal detection time to inhibit P-gp. Methods mRNA and protein expressions of P-gp in rats hippocampal tissue were detected by using immunohistochemistry , RT-qPCR and Western blot at different time points after modeling (0, 3, 6, 12, 24, 48, 72 h). Results The mean density of P-gp protein in the hippocampus of status epilepticus model was 0.325 1 ± 0.008 2 at 24 h, and was 0.396 3 ± 0.016 8 at 48 h, which were consistently higher than those of the control group (P < 0.05, P < 0.01, respectively). Results of qRT-PCR showed that MDR1a expression was significantly upregulated at 24 h and at 48 h (P < 0.05, P < 0.01, respectively). Western blot assay revealed that P-gp protein was also significantly increased at 48 h after seizures (P < 0.05). Conclusions The upregulation of P-gp after SE peaked at 48 h, which maybe the optimal detection time to detect drug resistant after SE.

Objective To propose a concept of irreducible tibial shaft fractures and to discuss their imaging characteristics and clinical significance.Methods A retrospective study was performed in 21 patients with tibial shaft fracture who had received intraoperative intramedullary nailing after limited open reduction at Department of Orthopaedics,The Second Affiliated Hospital to Anhui Medical University from November 2013 to June 2018.They were 14 males and 7 females,aged from 21 to 66 years (average,34.9 years).There were 15 left and 6 right sides.Firstly,closed reduction was performed followed by traction,folding and rotation,but repeated attempts failed to achieve smooth reduction or insertion of guide wire.Next,local limited open reduction had to be performed for intramedullary nailing.The X-ray and CT images of the tibial fractures were collected to analyze their imaging characteristics.The imaging manifestations were characterized into 4 types:single-segment type with intact fibula,multiple-segment type,interlocking type where the distal and proximal ends interlock commonly seen in short spiral and short oblique fractures,and incarceration type where the fracture interspace is blocked by a bone fragment.The therapeutic efficacy was evaluated at the final follow-up by knee scores of The Hospital for Special Surgery (HSS) and Kofoed ankle scores.Results Of the 21 patients,2 were single-segment type,4 multiple-segment type,13 interlocking type and 2 incarceration type.They were followed up for 7 to 50 months (average,22.7 months).The fractures united after 5 to 16 months (average,7.3 months).Postoperative knee pain was observed in 3 cases and delayed fracture union in 2.Osteomyelitis,superficial wound infection,implant breakage or malunion occurred in none of the patients.The therapeutic efficacy evaluated at the final follow-up by HSS knee scores and Kofoed ankle scores revealed 15 excellent,4 good and 2 fair cases,yielding an excellent to good rate of 90.5％.Conclusion The concept of irreducible tibial shaft fractures may lead to preoperative awareness on the part of the surgeons so that ineffective repeated reductions can be spared and the damage to the blood supply to the fracture ends and the operation time can be reduced.

Objective:To explore the short-term therapeutic outcomes of femoral neck fractures fixated by Femoral Neck System (FNS).Methods:A retrospective analysis was conducted of the 16 patients with femoral neck fracture who had been admitted to the Department of Traumatology & Orthopedics, The Second Affiliated Hospital to Anhui Medical University from January to December 2019. They were 6 males and 10 females, aged from 24 to 69 years (average, 47.5 years). According to the Garden Classification, there were 3 cases of type Ⅱ, 7 cases of type Ⅲ, and 6 cases of type Ⅳ. All the 16 patients were fixated with FNS. Recorded were operation time, frequency of intraoperative fluoroscopy, intraoperative blood loss, quality of postoperative fracture reduction, hospital stay, fracture healing time, and efficacy and complications at the last follow-up.Results:The 16 patients were followed up for 5 to 16 months after operation (average, 9.3 months). According to the Garden indexes, fracture reduction was evaluated as satisfactory in all the 16 patients. The operation time in this group ranged from 25 to 115 min with an average of 41.5 min, the frequency of intraoperative fluoroscopy from 14 to 47 times with an average of 26 times, the intraoperative blood loss from 35 to 210 mL with an average of 76 mL, the hospital stay from 3 to 9 days with an average of 4.6 days, and the fracture healing time from 3 to 6 months with an average of 4.5 months. By the Harris hip scores at the last follow-up, the function of the affected hip was rated as excellent in 10 cases, good in 5 and fair in one. Follow-ups revealed no complications like infection or implant loosening.Conclusions:The FNS is a new type of simple, reliable, safe and minimally invasive internal fixation for the treatment of femoral neck fractures. The plate, bolt, anti-rotation screws and locking screws in FNS can be inserted through a single minimally invasive incision, leading to reliable fixation, limited irritation to the lateral muscular soft tissues, and fine short-term outcomes.

Objective:To compare of the efficacy of Steinman pin-assisted and manual reduction for the treatment of distal femoral fracture with anterograde intramedullary nail.Methods:From January 2014 to August 2018, data of 54 patients with distal femoral fracture were retrospectively analyzed. According to the fracture reduction methods, patients were divided into two groups: bare-handed reduction group and Steinman pin-assisted reduction group (referred to Steinman pin group). There were 16 males and 10 females in bare-handed reduction group, with age of 37.5±9.2 years (range, 21-59 years). According to AO/OTA classification, 16 cases were type 32-A, 7 type 32-B, 3 type 32-C. There were 19 males and 9 females in Steinman pin reduction group, with age of 36.4±9.8 years (range, 18-55 years). According to AO/OTA classification, 19 cases were type 32-A, 7 type 32-B, 2 type 32-C. The reduction time, fluoroscopy times, intraoperative blood loss, fracture healing time, and knee joint function score of American hospital for special surgery (HSS) were compared between the two groups.Results:The amount of blood loss during operation was 142.78±29.76 ml in the bare-handed group, and 94.81±17.71 ml in the Steinman pin group. The reduction time of fracture was 14.19±2.50 min in the bare-handed group and 5.02±1.69 min in the Steinman pin group. The times of fluoroscopy during reduction was 12.56 ±2.01 in the bare-handed group and 5.01±1.51 in the Steinman pin group. There were significant differences in the above indexes ( t=12.19, 4.02, 5.47; all P < 0.05). All 54 patients were followed up for 12-51 months, with an average of 23.4 months. All the fractures healed, and there was no delayed union or nonunion. The healing time was 7.01±1.15 months in the bare-handed group and 5.99±0.97 months in the Steinman pin group. There were no significant difference. The HSS score of knee joint function was 23.7±4.1 before operation, 61.3±4.5 at 1 month after operation, 70.2±4.2 at 2 months after operation, 78.9±5.9 at 3 months after operation, 87.9±4.6 at 6 months after operation, and 93.1±5.8 at 12 months after operation, in the bare-handed group. Meanwhile, in the Steinman pin group, 22.5±3.8 before operation, 62.2±5.1 at 1 month after operation, 69.1±4.7 at 2 months after operation, 79.2±4.3 at 3 months after operation, 88.6±5.3 at 6 months after operation, and 92.3±6.1 at 12 months after operation. There were no significant difference between the two groups ( t=1.113, 0.689, 0.908, 0.212, 0.519, 0.494, P > 0.05). There were significant differences between the two groups at each time point before and after operation ( F=716.42, 815.52, P < 0.001). There were no complications such as injection point infection, vascular and nerve injury, failure of internal fixation and so on. Conclusion:Both groups had good functional recovery after operation. However, compared with bare-handed reduction, Steinman pin groupreduction has less intraoperative blood loss, shorter reduction time and less fluoroscopy times, which is a safer reduction method.