[Objective]To study changes of ultrastructures and respiratory function of the spinal cord mitochondrial in experimental spinal cord injury.[Method]Forty-eight SCI models in adult SD rats were built based on modified Allen's method.Mitochondria was extracted from injuried spinal cord tissue by using modified Estabrook's method at 6h,12h and 24h after SCI.Then ultrastructural changes and respiratory function,including R3,R4,RCR,P/O were observed.[Result]Ultrastructure of mitochondria was damaged greatly.The results revealed that R3、RCR and P/O of injuried spinal cord mitochondria decreased significantly after injury compared to the normal control group(P

Studies have shown that women's menopause caused by permanent cessation of ovarian function is closely related to lipid metabolism disorders. Er-xian Decoction has been used in the clinical treatment for gynecological diseases and has a good effect on diseases related to reduced sex hormone function. In this study, metabolomics was performed on bilateral ovariectomized model rats within 12 weeks after modeling to mimic the physiological state of menopausal women in different menopausal stages and Er-xian Decoction dosed model rats. The results of liver oil red O staining sections showed lipid metabolic disorder of bilateral ovariectomized model rats and the regulating effects of Er-xian Decoction. 46 potential biomarkers (6 steroid hormones, 3 sphingolipids, 11 phospholipids and 26 glycerides) in plasma and 32 potential biomarkers (1 steroid hormones, 20 phospholipids and 11 glycerides) in liver were obtained based on lipidomics analysis. Then, we analyzed the differential metabolic pathways and construct the lipid metabolism network significantly regulated by Er-xian Decoction. The results provided valuable information for in-depth understanding of the gradual changes on lipid metabolism disorders under menopausal conditions and the characteristics and mechanisms of compound Er-xian Decoction's regulatory effects. The study complied with the procedures established by the Animal Experiment Ethics Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences and passed the animal experiment ethics examine (No. 00000918).

BACKGROUND: Hippocampus is an important region involved in learning and memory. Some methods for freshly isolated hippocampal pyramidal neurons have been described. These methods need multiple enzymes or the procedure is complex.OBJECTIVE: To acquire large quantities of hippocampal neurons from rats suitable for patch clamp study.DESIGN: An animal experimental observation.SETTING: Department of Physiology and Pathophysiology, School of Medicine, Xi'an Jiao Tong University.MATERIALS: The experiments were carried out in the Medical Experimental Center, School of Medicine, Xi'an Jiao Tong University between March and October 2004. The animals were the 10-15-day-old SpragueDawley rats, bothmale and female.METHODS: Hippocampal pyramidal neurons were freshly isolated. Delayed rectifier potassium current and voltage-gated Ca2 + currents were recorded by whole-cell patch clamp configuration.MAIN OUTCOME MEASURES: Acute isolated neurons were observed with a Leica microscope. Recordings of delayed rectifier potassium currents and voltage-gated Ca2+ currents were obtained.RESULTS: ① Under the inverted microscope, the acute isolated neurons had a smooth and glassy surface, a three-dimensional contour, a pyramidal shape with a longer apical dendrite and several basal dendrites. ② After the whole cell recording configuration was formed, voltage-gated barium currents through calcium channels were elicited by 200 ms depolarizing potential from -70 mV to +20 mV, with an increment of 10 mV, a holding potential was -90 mV. Delayed rectifier potassium current (Ikd) was elicited by a protocol consisting of a multiple depolarizing pulses (-60 mV to +50 mV),preceded by a single prepulse step to -50 rmV, with inactivated transient outward potassium currents from a holding potential of -80 mV.CONCLUSION: This method is simple and ideal for the dissociation of neurons from rat hippocampus, and offers a powerful tool for functional analysis of ion channels by patch clamp.

Objective To culture the rabbit bone marrow derived mesenchymal stem cells (BMSCs) ,and to observe their biologi-cal characteristics in vitro and the expression of BMP2 and EGFP after Ad-EGFP-BMP2 infected on them .Methods To isolate and cultivate BMSCs by density gradient centrifugation and adherent culture in vitro ;the cell cycle and the surface marks were detected by flow cytometer ;rabbit BMSCs were differentiated into the direction of osteogenetic cells by in vitro induction .After transfection , the expression of exogenous gene in the cells was detected by the immunocytochemical staining and Western blot .Results About 56 .84% of rabbit BMSCs cell cycle was in the G1 phase;the D44 expression was positive and the CD45 expression was negative ;af-ter induction by osteogenetic cells ,the Ⅰtype collagen immunohistochemical staining was positive and the alizarin red staining was positive .After transfection ,the strong expression of BMP2 and EGFP in cells was showed by Western blot and the immunocyto-chemical staining .Conclusion rabbit BMSCs are successfully isolated ,cultured and identified to have the ability of osteogenetic dif-ferentiation ;the structured Ad-BMP-2/EGFP can efficiently transfected rabit BMSCs and stably express the target gene of BMP 2 .

Objective To study the effects of short-term cryopreservation on the proliferation ,bio-characteristics and osteogenic capability of rabbit BMSCs and lay the foundation for the further study .Methods The 5th generation of rabbit BMSCs were cryo-preservated by liquid nitrogen for 30 days ,and then thawing the cells to culture it to the 10th generation in vitro ,put the non-cryo-preservated and the same generation BMSCs as the control group .The MTT ,cell cycle ,cell surface marker ,the content of ALP and the immunohistochemical staining for collagen typeⅠ and alizarin red staining for calcium after differentiation induced by osteogene-sis were used to evaluate the proliferation ,bio-characteristic and osteogenic differentiation capability of rabbit BMSCs .Results The growth incubation period of BMSCs after cryopreservated was extended ,but it gradually recover through serial passage .The prolif-eration index and the proportion of BMSCs in G1 phase were 42 .9% ± 3 .4% and 57 .0% ± 3 .4% respectively .The positive rate of cell surface marker of CD44 was 93 .62% ± 1 .05% without expression of CD45 .The contents of ALP(U/gprot) were 6 .73 ± 1 .92 and 15 .99 ± 4 .36 in BMSCs after 7th day and 14th day with osteogenic induction ,respectively .The collagen typeⅠ and alizarin red staining for calcium indicated positive at BMSCs after 14th day and 21th day with osteogenic induction ,respectively .These results showed no significant difference compared with the control group (P>0 .05) .Conclusion Short-term cryopreservation has no obvi-ous impacts on the proliferation ,bio-characteristic and osteogenic capability of BMSCs and it could be used for further study .

Neurogranin, a neuron-specific postsynaptic protein, has been considered to play an important role in synaptic plasticity and learning and memory. The present study aimed to investigate the effects of prenatal restraint stress on neurogranin expression in rat offspring hippocampus. Pregnant rats were given a restraint stress (3 times a day for 7 d, 45 min each time) at the late stage of gestation except that in the control group. The offspring rats were divided into four groups: female control group, male control group, female stress group and male stress group. Expression of neurogranin was determined by immunohistochemistry and Western blot. The results showed that neurogranin-positive immunostaining was detected in all areas of the hippocampus. The staining density was stronger in the CA1 and CA3 regions than that in the dentate gyrus (DG) region. Western blot assay showed that neurogranin protein level in female and male prenatal stressed offspring was significantly lower than that in the controls (P<0.01). Neurogranin level was significantly lower in the female stress group than that in the male stress group, whereas there was no significant gender difference in the control group. Immunohistochemical data further confirmed these results. The present study provides evidence that prenatal restraint stress induces gender-dependent decrease in neurogranin expression in the offspring hippocampus. The prenatal restraint stress-induced decrease in neurogranin expression in the hippocampus might be associated with the deficit in spatial learning and memory reported previously.
Animals ; Blotting, Western ; Female ; Hippocampus ; chemistry ; Immunohistochemistry ; Male ; Neurogranin ; analysis ; Pregnancy ; Prenatal Exposure Delayed Effects ; Rats ; Rats, Sprague-Dawley ; Restraint, Physical ; Stress, Psychological ; metabolism

OBJECTIVETo study the diagnosis and treatment of the syndrome of inappropriate antidiuretic hormone secretion (SIADH) after brain injury.

METHODSA retrospective analysis was conducted on 12 patients suffered from SIADH after brain injury. The clinical features of these patients were similar to those of common hyponatremia. Most of the hyponatremia were detected by routine examinations. Supplement of salt as the initial treatment was used in these patients. If natremia did not rise or descended 2-3 days after treatment, SIADH was considered or diagnosed. Treatment scheme should be adjusted to limit water and natrium instead of supplying salt. Frusemide and albumin were the first choice for dehydration therapy.

RESULTS24-48 hours after limiting water and natrium, 12 patient's natremia level was back to normal. Eight out of 12 patients were corrected completely in 1 week, 1 in 14 days, and 1 in 3 months after injury.

CONCLUSIONSDiagnosis of SIADH is difficult before treatment though effective treatment can be obtained if we adopt correct strategy. In these patients, the diagnosis of SIADH is confirmed in the course of treatment.

Adolescent ; Adult ; Brain Injuries ; complications ; Female ; Humans ; Hyponatremia ; complications ; Inappropriate ADH Syndrome ; complications ; therapy ; Male ; Middle Aged ; Retrospective Studies ; Sodium ; therapeutic use ; Water ; administration & dosage

Objective To investigate the advantages of pediatric electronic fiber bronchoscope ( FBO) in the infant bronchial foreign body,discuss the clinical features of infant bronchial foreign body,lung imaging characteristics and the kinds of microscopically position,the change of airway mucosa after stimulation by for-eign body under local anesthesia in 30 cases of infant bronchial foreign body. Methods Thirty cases,aged 0 to 3 years,were collected from September to December,2014. All of them were with foreign bodies examined by FBO in pediatric bronchoscopy room in Shengjing Hospital of China Medical University. Results In all infants, 6 cases (20. 0%) without history of inhaled foreign bodies and 24 cases (80. 0%) with a record history of in-haled. In the aspects of signs:normal breath sounds with a history of no choking cough in children were 2 cases (6. 7%) ,wheezing sounds were 3 patients (10. 0%) and weakened side breath sound was 1 case (3. 3%);with a history of choking cough in children,6 cases(20. 0%) with normal breath sounds,12 cases(40. 0%) with wheez-ing,6 cases(20. 0%) with lateral breath sounds less. Lung imaging characteristics was lack of specific perform-ance:only a case of all(n=30)show foreign body directly. Otherwise,other 29 cases had no specificity. Lung em-physema in 13 cases (43. 3%) is the main characteristic,while normal imaging findings in 2 cases (6. 7%). For-eign bodies in 19 cases were in the left lung (63. 4%) and 21 cases(70. 0%) of foreign body stimulated granula-tion inside airway,necrosis sputum bolt in distal obstruction of airway occured in 5 cases (16. 7%). Inhalled time of foreign body in airway was 4. 5 [2. 8,12. 5] day and inhalled time of foreign body in airway correlation coefficient with granulation hyperplasia(r=0. 688,P=0. 000),there was a significant correlation. Main adverse reaction was low oxygen in 6 cases (16. 7%). Conclusion The diagnostic accuracy of FBO under local anesthesia on children is high-er than other methods,and the FBO bronchial foreign bodies under local anesthesia is a safe and effective method.

Objective To investigate the time relationship of the change,and diagnostic accuracy and sensitivity between retinal light threshold fluctuations (LTF) and retinal nerve fiber layer (RNFL) and ganglion cell complex(GCC) thickness on high-risk primary open-angle glaucoma (POAG).Methods Totally 319 patients (319 eyes) with high-risk in POAG from the First Affiliated Hospital of Kunming Medical Universityand during December 2009 and December 2017,50 healthy individuals (50 eyes) as control were collected in this longitudinal cohort study.Visual field and OCT were reviewed every 6 months on the high-risk group and every 12 months on the control group.High-risk groups inclusion criteria:vertical C/D≥0.6;early visual field defect (according to glaucoma visual field damage GSS2 quantitative grading standards,mean deviation and pattern standard deviation of central field exceeds the border as an early visual field defect);continuous repeatable results.The first field and OCT results in the absence of visual field defects and C/D≥0.6,which were conformed reliability indicators and removed learning effects as a baseline.When patients achieve POAG diagnosis criteria first time which was recorded as a turning point.And they were divided into early group meanwhile were ended of follow-up.After the last follow-up,the inspection data was segmented counted in yearly interval.The changes of LTF,thickness of RNFL and GCC during the follow-up period in the early POAG group and the control group were observed.The loss rate and change rate in each period were compared for the assessment of their trends with time.Followed by calculation of the area under receiver operating curves (AUC) to compare the predicted value of POAG and the sensitivity at 95％ specificity in each period.Results After last follow-up,totally 67 patients 67 eyes (early POAG group,37 males and 30 females) were entered the turning point.The mean follow-up of the early POAG group and the control group were 6.6 and 6.4 years.The average RNFL thickness was 79.05± 8.09 μm,GCC thickness was 71.58 ± 8.41 μm,LTF was -6.05 ± 7.02 dB in early POAG group.The average RNFL thickness was 93.49± 6.24 μrm,GCC thickness was 79.72± 6.32 μm,LTF was-0.31 ± 0.58 dB in the control group.The differences of LTF and the thickness of RNFL and GCC were statistically significant (t=-5.97,-10.42,-5.60;P＜0.001).The AUC of RNFL,GCC thickness and LTF increased with time in the early POAG group.The sensitivity was gradually increased at 95％ specificity:5th year before to at turning point,RNFL thickness AUC was 0.15,0.65,0.71,0.77,0.85,0.92,and sensitivity was 20％,56％,61％,65％,70％,76％,respectively;GCC thickness AUC was 0.12,0.53,0.69,0.74,0.82,0.90,and sensitivity was 14％,53％,69％,74％,82％,90％,respectively;the AUC of LTF was 0.10,0.21,0.33,0.75,0.86,0.91,and sensitivity was 7％,17％,44％,65％,78％,87％,respectively.Conclusions The earliest time of structural functional damage of POAG is at the 4th year before confirmed,simultaneous RNFL diagnosis accuracy and sensitivity are better than GCC and LTF.The earliest time of visual functional damage of POAG is at the 2th year before confirmed,simultaneous LTF diagnosis accuracy and sensitivity are better than RNFL and GCC.

Objective To improve the pediatrician's understanding of severe combined immunodefi-ciency disease (SCID),so as to strengthen the early diagnosis and treatment of SCID. Methods The clinical manifestations,related immunological findings,imaging findings and outcomes of 10 SCID children admitted to our hospital from 2007 to 2018 were retrospectively analyzed. Results The clinical manifestations of primary SCID were frequent infections shortly after birth. There were 8 males and 2 females in this study. The average age of onset was 4. 2 months,and the average age of diagnosis was 6 months. Eight cases of thymus shadow were absent,9 cases of pulmonary CT showed severe pneumonia,and 3 cases of pulmonary fungal infection. Six of 7 children died of infection and the age of death was less than 1 year old. Laboratory exami-nation showed that 10 patients had abnormal cell and humoral immune function;10 cases of CD3 +T cells were <20%,3 cases of CD16 +CD56 +( NK%) >85%,7 cases ≤ 2%. There was an increase in B cell reactivity of 7 cases,but the secretion of immunoglobulin in 5 cases was significantly reduced. Seven cases of IgG<2. 0 g/L,3 cases>2. 0 g/L. Eight cases were confirmed of the primary SCID by genetic testing. Con-clusion The clinical manifestations of SCID are frequent bacterial,viral and fungal infections in a short time after birth. In clinical work,when small infants are repeatedly infected after birth,or multiple sites are serious-ly infected and prolonged,we must think of the possibility of SCID and timely related immune function tests, strive to achieve early diagnosis,timely treatment,and early bone marrow stem cell transplantation.