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.

OBJECTIVETo observe the renal protection effects of Compound Shenhua Tablet (CST) on diabetic nephropathy (DN) rats.

METHODSDN rats were given a normal diet for 9 months after they were induced by intraperitoneal injection of STZ at the dose of 65 mg/kg after uninephrectomized. They were randomly divided into 4 groups, i. e., the normal control group, the model control group, the CST group, and the Irbesartan group. The intervention was given by gastrogavage for 6 weeks. The general state, 24 h urine protein, urine micro-albumin (mAlb), serum creatinine (SCr), blood urea nitrogen (BUN), glucose (GLU), triglyceride (TG), total cholesterol (TC), total protein (TP), and albumin (ALB) levels were observed before and after intervention. Renal pathological changes were observed by PAS staining and transmission electron microscope.

RESULTSAfter 6 weeks of drug intervention, when compared with the model control group, the general state was improved in the CST group and the Irbesartan group. The levels of 24 h urine protein, urine mAlb, SCr, BUN, GLU, TG, and TC were obviously lower in the CST group and the Irbesartan group than in the model group as well as in the same group before treatment (P<0.05, P<0.01). There was no statistical difference between the two treatment groups (P>0.05). The renal pathological changes and the renal ultrastructure were improved to some degree in the two groups when compared with those in the model control group.

CONCLUSIONSCST could attenuate the renal damage of diabetes and delay renal deterioration process. Its effectiveness was equivalent to that of Irbesartan.

Animals ; Diabetes Mellitus, Experimental ; drug therapy ; Diabetic Nephropathies ; drug therapy ; Drugs, Chinese Herbal ; pharmacology ; Kidney ; Male ; Phytotherapy ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo establish a culture method for a large amount of highly purified osteoclast-like cells in vitro. To investigate the gene expression of some osteoclast marker enzymes. To lay the foundation for the further study of the signal path on the differentiation and formation of osteoclast-like cells.

METHODSThe bone marrow mononuclear cells of rat were treated with 30 ng/mL macrophagecolony-stimulating factor (M-CSF) and 50 ng/mL receptor activator of NF-kappaB ligand (RANKL) and cultured for 6 days. After culturing, cells were evaluated by morphology observation, tartrate-resistant acid phosphatase (TRAP) staining, Giemsa staining, pit staining, and the gene expression of some osteoclast marker enzymes.

RESULTSThe TRAP-positive mononuclear cells were more frequently observed than the multinucleated cells and pit staining could be seen on the dentine slice. The transcription expression of TRAP, matrix metalloproteinase-9 (MMP-9), membrane-type1-matrix metalloproteinase (MT1-MMP) and cathepsin K were detected by RT-PCR.

CONCLUSIONThe cooperation of M-CSF and RANKL could induce a large amount of highly purified osteoclast-like cells formation in rat bone marrow culture. The typical characteristics of osteoclast-like cells were demonstrated and the enriched osteoclast-like cells expressed TRAP, MMP-9, MT1-MMP and cathepsin K.

Animals ; Bone Marrow Cells ; Cathepsin K ; Cell Differentiation ; Cell Line ; In Vitro Techniques ; Matrix Metalloproteinase 9 ; Osteoclasts ; RANK Ligand ; RNA, Messenger ; Rats ; Receptor Activator of Nuclear Factor-kappa B

OBJECTIVETo study molecular epidemiology of norovirus (NV) infections, stool specimens collected from children with acute diarrhea were tested by TaqMan real-time reverse transcription polymerase chain reaction (RT-PCR) for the viral specific nucleic acid segments.

METHODSFecal samples from a total of 1260 children who had watery diarrhea seen from December 2006 to December 2007 in Guangzhou were analyzed by real-time RT-PCR. The primers and probes used for rapid detection and typing of NV strain target NV sequences were at the ORF1-ORF2 junction, a highly conserved region of the NoV genome. The positive specimens were determined by nested PCR and sequenced.

RESULTSTotally 257 specimens were positive for NV with a positive rate of 20.40%. Shedding of NV type GI was detected in 6.90%, type GII in 16.98% respectively, while the positive number of mixed infection with GI and GII was 44. Of the NV strains that were cloned and sequenced, GI was GI-3, GI-2 and GI-4 detected in positive specimens respectively; meanwhile, GII-4 was most commonly seen in genome II, followed by GII-3 and GII-7. In addition, the average age of children infected with NV was less than 2 years. An epidemic occurred during the winter and early spring (December through the next March).

CONCLUSIONNV was one of the important pathogens for acute diarrhea among children in Guangzhou, which suggested GII-4 was the prevalent strain.

Caliciviridae Infections ; epidemiology ; Child, Preschool ; China ; epidemiology ; Diarrhea ; epidemiology ; etiology ; virology ; Feces ; virology ; Humans ; Infant ; Molecular Epidemiology ; Norovirus ; classification ; genetics ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

Objective To study the effects of ribosomal protein L41 (RPL41) on the proliferation and apoptosis of human retinoblastoma Y79 cells and its underlying mechanisms.Methods Y79 cells were seeded in RPMI 1640 medium containing 10％ fetal bovine serum for passage culture.Then the cells were divided into control group,with cells left untreatment,(40 μmol · L-1,80 μmol · L 1 and 120 μmol · L-1) RPL41 treatment group according to the concentration.Next CellTiter-Glo fluorescence cell viability testing system was used to observe the viability of Y79 cells in all groups,and flow cytometry was applied to measure the cell apoptotic rate in 100 μmol · L 1 RPL41 treatment group,with Hoechst staining for the observation of nuclear morphometry of apoptotic cells,and finally,Western blot was used to determine the expression of activating transcription factor 4 (ATF4) of each group.Results Compared with the control group,the viability of Y79 cells in the 40 μmol · L-1 RPL41 treatment group was (97.9 ± 1.5) ％,with no significant difference (P =0.055);and the viability in the 80 μmol · L-1 and 120 μmol · L-1 RPL41 treatment group was (87.6 ± 1.8)％ and (63.9 ± 2.0) ％,respectively,both of which were significantly different from the control group (both P ＜ 0.05),so RPL41 inhibited the viability of Y79 cells,and 100 μmol · L-1 RPL41 promoted the apoptosis of Y79 cells,with the apoptotic rate of (17.33 ± 2.47)％.Compared with normal cells,the apoptotic cells in the 100 μmol · L 1 RPL41 treatment group showed bright color and smaller cell volume by Hoechst staining.Western blot showed that PRL41 significantly decreased the expression of ATF4 protein and the expression of ATF4 protein in the 40 μmol · L 1,80 μmol · L-1 and 120 μmol · L 1 treatment group were 0.76 ± 0.04,0.29 ± 0.04,0.29 ± 0.05,respectively,all of which were significantly different from the control group (all P ＜ 0.01).Conclusion RPL41 can inhibit the proliferation and promote the apoptosis of human retinoblastoma Y79 cell,and its mechanism may be related to the expression of ATF4.

OBJECTIVETo study the prevention and treatment of postoperative diabetes insipidus after removal of pituitary tumor through transsphenoidal operation, to decrease the incidence of postoperative complications and improve the treatment of pituitary tumor.

METHODSThe clinical data of 86 cases of transsphenoidal resection of pituitary tumor in recent 8 years were retrospectively reviewed, including 35 endoscopic operation and 51 microscopic operation. The incidence, prevention and treatment of diabetes insipidus were statistically analysed.

RESULTSThere were 18 cases of postoperative diabetes insipidus in total of 86 operations, including 15 acute cases, 3 delayed cases. Twelve were temporary , which recovered within 1 week. After prompt treatment, 14 recovered within 1 week, 4 recovered within 2 weeks. No persistent diabetes insipidus was found.

CONCLUSIONSThe key points to prevent postoperative diabetes insipidus lay in the improvement of operative skills, careful protection during operation and avoidance of unnecessary injury. In case of diabetes insipidus occurred, rational use of antidiuretics and correction of electrolyte balance were effective in the treatment of postoperative diabetes insipidus.

Adult ; Diabetes Insipidus ; etiology ; Endoscopy ; Female ; Humans ; Male ; Middle Aged ; Pituitary Neoplasms ; surgery ; Postoperative Complications ; etiology ; Retrospective Studies ; Sphenoid Sinus ; surgery

Ginkgo terpene lactones, as an important active ingredient from Ginkgo leaves, has high medicinal values and has been widely used in clinics. This article would review the researches both at home and abroad, including chemical composition, structure-activity relationship, analytical methods, pharmacological effects, pharmacokinetic characteristics, and so on, providing a reference for further development and utilization of ginkgo terpene lactones.

OBJECTIVETo provide scientific basis data for revising the national hygiene criteria of "Classification of hazard conditions of productive dust" (GB5817-86).

METHODSThe data of the retrospective study and the field survey data were analyzed with correlation and regression analysis. The product of total dust concentration of respiratory exposure (mg/m(3)), total ventilation during exposure (m(3)/d per psrson), and level of free SiO(2) in dust (%) was the respiratory exposure dose of free SiO(2) (mg per day per person) which was used as dose criteria value of classification of hazard degree of dust.

RESULTSUsing free SiO(2) exposure dose and the dose-effect relationship, the hazard degrees of the dust were divided into 5 grades: 0, I, II, III, IV (0 - 8.0, 8.1 - 12.0, 12.1 - 16.0, > 24.1 mg per day per person).

CONCLUSIONThe exposure dose of free SiO(2) is closely related to the pathogenesis of silicosis. Using the exposure dose of free SiO(2) as the classification indicator of hazard degree of dust is reliable, simple and easy to execute.

China ; Dust ; analysis ; Hazardous Substances ; analysis ; Humans ; Lung ; diagnostic imaging ; pathology ; Occupational Exposure ; standards ; Radiography ; Retrospective Studies ; Safety Management ; standards

BACKGROUNDCisplatin (DDP) is one of most effective and most commonly used therapeutic agent in treating tumors, it can accumulate in the kidney and lead to acute renal failure. MicroRNA-181a can induce cell apoptosis by suppressing the expression of Bcl-2 family. In the present study, we investigated the role of microRNA-181a in the apoptosis of tubular epithelial cell induced by DDP.

METHODSHK-2 cells were cultured, transfected with microRNA-181a inhibitor for 48 hours, and stimulated with 50 µmol/L cisplatin for 24 hours. MicroRNA-181a expression was analyzed by real time PCR, and cell apoptosis was detected by flow cytometry. Moreover, Bcl-2 and Bcl-2-associated X protein (Bax) expression were measured by Western blotting.

RESULTSMicroRNA-181a expression significantly down-regulated in cells transfected with microRNA-181a inhibitor, compared with that in untransfectd cells (21.19 ± 2.01 vs. 38.87 ± 1.97, P < 0.05). Cell apoptosis induced by DDP significantly decreased in cells transfected with MicroRNA-181a inhibitor. Compared with DDP treated cells alone, Bcl-2 expression strikingly was up-regulated and Bax expression was down-regulated in cells transfected with microRNA-181a inhibitor.

CONCLUSIONOne pathway of DDP induces apoptosis of tubular epithelial cell by suppressing Bcl-2 expression is achieved by regulating the target gene of MicroRNA-181a.

Apoptosis ; drug effects ; genetics ; Blotting, Western ; Cell Line ; Cisplatin ; pharmacology ; Flow Cytometry ; Humans ; MicroRNAs ; genetics ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Small Interfering ; genetics ; Real-Time Polymerase Chain Reaction ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; bcl-2-Associated X Protein ; metabolism

Objective To establish the systemic lupus erythematosus (SLE) mouse model through pristaneip injection and validate the model comprehensively.Methods Female BALB/c mice of 6-8 weeks were randomly divided into two groups.Animals in model group were injected with 0.5 mL pristane by ip injection while in control group with 0.5 mL normal saline.Anti-systemic lupus erythematosus antibodies (anti-SLE) and anti-double strand DNA antibodies (anti-dsDNA) were checked before injection and monthly thereafter.Proteinuria was detected before injection and every month thereafter.All mice were bled to death 6 months after injection.Kidneys were excised to observe the histopathologic evidence of glomerulonephritis.Results The concentration of anti-dsDNA and anti-SLE antibody in sera was higher of model group than that of control group two months after pristane injection,and the concentration of antibody gradually increased within 6 months.At the sixth months,the protein concentration of urine in most model group mice was ++++.The histopathology and imunoflorescence of kidney sections indicated typical evidence of glomerulonephritis in model group.Conclusion The murine lupus model can be successfully established in female BALB/e mouse with a single ip injection of 0.5mL ofpristane.