Abstract BACKGROUND: Recently, microcarrier culture as a high concentration cel culture technology has been used in hepatocyte culturein vitro. OBJECTIVE:To periodicaly observe the morphological changes of human hepatocyte L-02 cultured by the spherical porous chitosan microcarrier. METHODS:The self-made spherical porous chitosan microcarrier sample as a support to culture human hepatocytes L-02 acted as experimental group; non-spherical porous chitosan microcarrier as a support to culture human hepatocytes L-02 as control group. Cels in two groups underwent cel count at regular time, and the morphological changes were observed in the experimental group, including inverted phase contrast biological microscope observation and scanning electron microscope observation. RESULTS AND CONCLUSION:The quantity of cultured cels in the two groups was increased in the first 3 days and reached the peak at the 3rd day. The cel quantity in the experiment group was obviously higher than that in the control group (P < 0.05). There was no significant difference in the three samples of the experimental group (P > 0.05). The quantity of hepatocytes adhered to the microcarrier surface was gradualy increased in the former 3 days under the inverted phase contrast biological microscope. There were lots of cel clusters on the surface of a greater part of microcarriers, with the total survival rate of more than 90%, and hepatocytes kept a good morphological structure. Under the scanning electron microscope, lots of hepatocytes adhered tightly to each other on the surface and section of microcarrier as wel as inside the microcarrier. It is indicated that the self-made spherical porous chitosan microcarrier as a support in three-dimensional environment can undergo cel culture with high concentration.

Objective The aim of this article is to study the clinical features, image, electroencephalogram and gene mutation of mitochondrial encephalomyopathy with MELAS and to improve understanding of the disease. Methods Clinical data was collected from 12 MELAS cases. Their clinical symptoms, head imageological result,electroencephalo-gram and gene mutation were analyzed. Results The average onset age was 28 years old. The main manifestations of 12 patients were epilepsy(66.7%), stroke-like episodes(50.0%), mental behavious disorder and decrease of perceive (41.7%),diabetes or abnormal glucose tolerance(41.7%), hearing impairment(41.7%). The imaging results showed long T1, long T2 and high signals on DWI. The nidus involved occipital lobe, temporal lobe, parietal lobe most. Five of six cases undergoing MRS imaging had large lactate peak at 1.33 ppm. There were 9 cases carrying mtDNA A3243G muta-tion. Conclusion The clinical presentation of MELAS is highly variable without any obvious specificity on MRI and EEG. Large lactate peak on MRS may indicate the disease. Most patients carry A3243G mutation.

Gastrointestinal Tract ; physiopathology ; Humans ; Infection ; physiopathology ; Sepsis ; physiopathology ; Shock, Septic ; physiopathology

Objective To investigate the effects of postoperative sleep deprivation on the expression of choline acetyltransferase (ChAT) in hippocampi of aged rats.Methods Forty-eight male Wistar rats,aged 20 months,weighing 500-600 g,were randomly divided into 4 groups (n =12 each) using a random number table:control group (group C) ; operation group (group O) ; sleep deprivation group (group S) ; postoperative sleep deprivation group (group OS).Sleep deprivation was induced in the rats by housing them on small platforms over water.They fell into the water if they lost muscle tone.All the rats had free access to food and water.In group OS,splenectomy was performed,and all the rats underwent 24 h sleep deprivation after the rats were awake.All the rats underwent 24 h sleep deprivation at the corresponding time point in group S.Morris water maze test was carried out at 24 h after operation.The number of ChAT positive cells in the hippocampal CA1 region was counted after completion of Morris water maze test.Results Compared with group C,the escape latency was significantly prolonged,the time of staying at the original platform quadrant was shortened,and the frequency of crossing the original platform and the number of ChAT positive cells in the hippocampal CA1 region were decreased after operation in O and OS groups,and no significant changes were found in the parameters mentioned above in group S.Compared with group O,the escape latency was significantly prolonged,the time of staying at the original platform quadrant was shortened,the frequency of crossing the original platform was decreased,and there was no significant difference in the numberof ChAT positive cells in the hippocampal CA1 region in group OS,and no significant changes were found in the parameters mentioned above in group S.Compared with group S,the escape latency was significantly prolonged,the time of staying at the original platform quadrant was shortened,and the frequency of crossing the original platform and the number of ChAT positive cells in the hippocampal CA1 region were decreased after operation in group OS.Conclusion The mechanism by which postoperative sleep deprivation induces cognitive decline is not related to the expression of ChAT in hippocampi of aged rats.

Objective To investigate the dynamic changes in peripheral blood of patients with acute pancreatitis α-MSH,TNF-α,PCTS and the value of the patient assessment of disease severity.Methods Emergency surgery in the Second Affiliated Hospital of Anhui Medical University from March 2011 to March 2014 were treated 68 cases of acute pancreatitis patients,36 patients with mild,32 cases of severe group.All patients were extracted 1,3,5,7 days after admission blood,and α-MSH was measured by enzymelinked immunosorbent assay,the content of TNF-αand PCT.Results Compared with the mild group,the serum of patients with severe group of α-MSH concentrations were lower (P ＜ 0.05),while serum TNF-αand PCT levels higher than the mild group (P ＜ 0.05);in disease severity assessment,α-MSH sensitivity of 82.35％ and a specificity of 88.24％,the accuracy was 87.5％,TNF-α sensitivity of 76.47％ and a specificity of 82.35％,the accuracy was 81.25％,PCT sensitivity of 70.59％ and a specificity of 76.47％,75.00％ accuracy,PCT + TNF-α + α-MSH sensitivity of 88.24％ and a specificity of 94.12％,93.75％ accuracy;area under the PCT,TNF-α,α-MSH and the combination of the three curves 0.690,0.696,0.768 and 0.885,respectively.Conclusions Serum α-MSH,monitoring TNF-α,PCT levels have some clinical value in patients with acute pancreatitis disease severity assessment,including α-MSH in the condition assessment of the sensitivity,specificity and positive predictive value were higher than the latter two,and peripheral α-MSH in patients with a low level indicates a serious condition,poor prognosis;addition,combined detection with higher clinical value in the assessment of disease severity.

OBJECTIVETo study the effects of modified Mahuang Fuzi Xixin Decoction extract (MFXDE) on ischemia/reperfusion induced atrioventricular (A-V) block in rabbits model.

METHODSNew Zealand rabbits were randomly divided into six groups, three test groups (Group TA, TB, TC) and three control groups (Group CA, CB, CC), 10 in each. MFXDE 2 mL/kg was given twice a day by gastrogavage to the test groups, while to the control groups, equal volume of normal saline was given instead, for 3 successive days. Twenty minutes after the last gastrogavage, right coronary artery ligation was performed in rabbits under anesthesia with 1.2 g/kg of 20% urethane via ear marginal vein injection, and lasted for 15 min (Group TA and CA), 60 min (Group TB and CB), and 120 min (Group TC and CC), respectively. ECG lead-II and His Bundle ECG were recorded at different time points to observe P-R interval and A-H interval.

RESULTSP-R interval and A-H interval in the test groups were shorter than in the control groups significantly (all P < 0.05) at the time point of 5 min after ischemia; and at the reperfusion stage, a re-extending phenomenon of P-R and A-H could be found in the CC group after 60 min reperfusion (P < 0.05), but it didn't occure in all the other test groups.

CONCLUSIONMFXDE could improve the ischemia/reperfusion induced A-V conductive function.

Animals ; Atrioventricular Block ; drug therapy ; etiology ; physiopathology ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Male ; Myocardial Reperfusion Injury ; complications ; Rabbits ; Random Allocation

Abstract?AlM:To evaluate the accuracy and practicability of three - dimensionally preformed Arbeitsgemeinschaft Osteosynthese AO titanium mesh plates for orbital fractures.?METHODS:Forty-seven patients with isolated blow-out orbital fractures were included in this study. Fracture locations were as follows: floor/medial wall ( n = 26, 55%), medial wall ( n = 12, 26%), and floor ( n = 9, 19%) . The floor fractures were exposed by a standard transconjunctival approach, whereas a combined transcaruncular transconjunctival approach was used in patients with medial wall fractures with temporary dissection of inferior oblique muscle. A three -dimensionally preformed AO titanium mesh plate was selected according to the size of the defect previously measured on the preoperative computed tomographic scan examination and fixed at the inferior orbital rim with 2 screws. The accuracy of plate positioning of the reconstructed orbit was assessed on the postoperative computed tomography ( CT) scan. The practicability of clinical use of AO three - dimensionally preformed titanium mesh plates was assessed on the preoperative and postoperative clinical data.?RESULTS: Postoperative orbital CT scan showed an anatomic three- dimensional placement of the orbital mesh plates in all of the patients. All patients had a successful treatment outcome without clinical complications. 40 patients ( 87%) had a successful enophthalmos correction. 25 patients ( 86%) had a successful recovery from diplopia.?CONCLUSlON: Three- dimensionally preformed AO titanium mesh plates for orbital fracture reconstruction results in an accurate anatomic restoration of the bony orbital contour with a high rate of success to correctenophthalmos and diplopia.

Background Pupillary light reflex has been widely used in the diagnosis and evaluation of visual system and nervous system diseases.However,in animal experiments,functional evaluation of the visual system and nervous system needs more advanced technology and are affected by many factors.Objective This study was to explore the use of the dynamic pupillometer in evaluating pupillary light reflex and to discuss the influence of brightness of stimulate on relevant curve parameters in C57BL/6 mouse.Methods Ten healthy SPF male C57BL/6 mice were collected in this experiment.White light of five luminance levels (2,8,32,128,256 cd/m2) was used to stimulate the mice following a 2-hour dark adaptation.The stimulation was given at the 60-second intervals,for a duration of 100 ms at every stimulation.An infrared camera and video capture card were used to capture digital images during the measuring process in a scotopic environment,at a speed of 60 frames per second.Measuring outcome was saved in the*.AVI format.A software that was developed by our group was used to determine pupil diameter and output pupillary light reflex curve offline.Pupil initial diameter (R1),constriction amplitude (CA),constriction velocity (CV),latency (T1),time for maximum velocity (T2),time for maximum constriction (T3),time for maximun con-striction to 10.1％ R1 re-dilation (RT)and re-dilation velocity (RV)were assessed,and the correlations between luminosity and measuring parameters were analyzed using the Spearman rank correlation.The use of animals followed the Regulations for thd Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results R1 values showed no statistically significant difference among the 5 different luminosity groups(F=1.117,P=0.361).A positive linear correlation was found between stimulating luminosity and CA(r=0.508,P＜ 0.01),but negative correlations were seen between stimulating luminosity and CV or RV (r=-0.625,-0.609,P＜0.01).T1 and T2 values in the 5 different luminosity groups were not statistically significant (F =0.202,P =0.936 ; F =1.584,P =0.195).The different levels of stimulating luminosity showed positive linear correlations with T3 and RT values (r =0.791,0.609,P＜ 0.01).Conclusions The dynamic pupillometer can quantitatively measure the pupillary light reflex of C57BL/6 mice.The pupillary light reflex dynamic curve parameters of mouse were affected by stimulus luminosity levels.These outcomes offer a basis for the application of the dynamic pupillometer system for measuring pupillary light reflex in animal models.

Objective To observe whether calreticulin-induced mitochondrial dysfunction is involved in cardiomyocyte hypertrophy induced by angiotensin Ⅱ (AngⅡ).Methods Primary culture of neonatal rat cardiomyocytes was further confirmed by immunocytochemistry.The cardiomyocytes were randomly divided into model group,valsartan group and control group.The model group was subdivided into three groups which were separately treated with 1 0-8 mmol/L, 1 0-7 mmol/L, and 1 0-6 mmol/L Ang Ⅱ. Calreticulin expression, mitochondrial membrane potential level, enzyme activities, cell surface area and protein synthesis rate were observed.Results Cell surface area and protein synthesis rate were both increased in model groups compared with control group.Mitochondrial membrane potential level and enzyme activities were lower in model groups than in control group,while calreticulin expression was up-regulated.Pretreatment with valsartan partially reversed all the above changes.Conclusion Mitochondrial dysfunction induced by calreticulin may be an important mechanism of myocardial hypertrophy.

Objective To evaluate the forecasting capability of serum N-terminal pro-brain natriuretic peptide ( NT-proBNP) level for Left ventricular ( LV) remodeling of patients with acute myocardial infarction (AMI) through observation of the relationship between serum NT-proBNP on the 3rd day and LV remodeling following onset of AMI. Methods Electrochemilumin-escence was adopted to determine the serum NT-proBNP level on the 3rd day after AMI attack for 106 cases of patients with anterior, anteroseptal and anterolateral AMI, with echocardiography performed to detect LV end-diastolic diameter (LVDd) and leftventricular ejection fraction (LVEF) respectively on the 3rd day and 3 months after the attack. Results The serum NT-proBNP level of AMI patients on the 3rd day averaged 1039. 28 (241. 50-1 184. 25) ng/L For AMI patients on the 3rd day and 3 months after, LVDd rose to (53?7) mm (P 0.05) from (53?8) %. NT-proBNP concentration on the 3rd day after AMI onset was positively correlated with△LVDd significantly, r =0. 403 (P 5mm during the 3 months and LVEF at the 3rd month≤40 % , AUC was 0. 893. Conclusion The test results showed serum NT-proBNP level for AMI patients on the 3rd day may be used as one of forecasting indexes of LV remodeling for AMI of advanced stage.