Objective To evaluate the effect of anisodamine on endoplasmic reticulum stress during acute kidney injury (AKI) in rats.Methods Forty-two nale Sprague-Dawley rats,weighing 200-220 g,were randomly divided into 3 groups:control group (group C,n =6),AKI group (n =18) and anisodamine group (group AD,n =18).AKI was induced by intramuscular injection of 50％ (v/v) glycerol 10 ml/kg into bilateral hind limbs in groups AKI and AD.In addition,anisodamine 10 mg/kg was injected intraperitoneally 20 min before intramuscular injection of glycerol in group AD.In group C the rats received intramuscular injection of normal saline 10 ml/kg into bilateral hind limbs.Six rats were chosen immediately after injection of normal saline in group C or at 1,6 and 24 h after glycerol injection in groups AKI and AD and then anethetized with intraperitoneal pentobarbital.The animals were sacrificed and kidney specimens were obtained and cut into sections which were stained with hematoxylin and eosin for pathological examination.The pathological changes of the renal tubules were scored.The expression of glucose-regulated protein 78 (GRP78) and oxygen-regulated protein 150 (ORP150) in renal tissues was determined by immuno-histochemistry and Western blot.Results Compared with group C,the pathological scores were significantly increased and the expression of GRP78 and ORP150 was up-regulated at all time points in groups AKI and AD (P ＜ 0.01).Compared with group AKI,the pathological scores were significantly decreased and the expression of GRP78 and ORP150 was down-regulated at all time points in group AD (P ＜ 0.05 or 0.01).Conclusion Anisodamine can ameliorate AKI through inhibiting endoplasmic reticulum stress in renal tubular epithelial cells and decreasing endoplasmic reticulum stress-induced cell apoptosis in rats.

Aim To investigate the Yiqi Yangyin Qingre Oral Liquid(YYQ) on insulin resistance in hyperglycemic fatty rats.Methods The model of hyperglycemic fatty rats was induced by intravenous injection with a low dose of streptozotocin(STZ) and a fed high fat and high caloric diet.The YYQ was given via gavage for a period of 4 weeks.The fasting blood glucose(FBG),oral glucose tolerance(OGT),serum insulin(INS),triglyceride(TG),total cholesterol(TC),free fatty acid(FFA)levels,glucose infusion rate(GIR) by euglycemic hyperinsulinemic clamp technique as well as erythrocyte insulin receptors(INSR)of rats were tested when the experiment was finished.The weight of the epididymal fat pads and the perirenal fat pads were measured.The IR,IRS-1mRNA expressions in liver tissue were measured by RT-PCR technology.Results YYQ treatment decreased the levels of FBG,TC,TG,LDL-C,FFA and increased the level of HDL-C in hyperglycemic fatty rats.As compared with the model rats,glucose tolerance was improved and body weight was reduced in hyperglycemic fatty rats treated with YYQ.Moreover,YYQ treatment upregulated IR,IRS-1mRNA expressions in liver tissue and increased insulin receptor amounts on the erythrocytes surface as well as GIR in the hyperinsulinemic euglycemic clamp experiment.Conclusions YYQ ameliorated insulin resistance in hyperglycemic fatty rats,and its mechanism might be related to increment of INSR and stimulation of insulin pathway.

OBJECTIVETo explore clinical significance of rotational axis of distal femur on Chinese adults in total knee arthroplasty (TKA).

METHODSThere were 86 Chinese adults (106 normal knees) including 47 males (53 knees) and 39 females (53 knees), 54 knees were on left and 52 on right. The CT scan was employed in the distal femur. The scan direction was aligned to be on the plane perpendicular to the mechanical axis of the femoral. The CT images of cross sections across lateral and medial femoral epicondyle were moved to personal computer,lateral angle between anterior posterior line (APL) and surgical transepicondylar axis (STEA) (ATA),lateral angle between posterior condylar line (PCL) and APL (APA), angle between perpendicularity of APL and PCL (A-PA), posterior condylar angle (PCA), condylar twist angle (CTA), angle between clinical transepicondylar axis (CTEA) and STEA (CSA) were measured. These values were divided into different groups according to gender and side, the values of CTA, PCA, A-PA, angle PT (varus of tibia plateau), constant 3, ATA, APA and constant 90° were compared by statistically. A-PA and PCA, and CTA were analysed statistically with the liner regression, the relationship among CTEA, STEA ,PCL, APL and PLP were performed to assess by liner regression.

RESULTSATA was (89.79 ± 1.22)°, APA was (84.84 ± 1.83)°, A-PA was (5.16 ± 1.83)°, PCA was (4.80 ± 1.23)°, CTA was (8.23 ± 1.40), CSA was (3.45 ± 0.68)°. All the parameters had no differences on sex and side,but CSA had difference on male and female. There was no difference among angle PT, PCA, A-PA. There was significant difference in CAT, constant 30 and angle PT, PCA,A-PA. There was no difference between ATA and constant 90°, but there was difference between APA and constant 90°. There was relativity between PCA and CTA, and also PCA and A-PA, CTA and A-PA. There was significant relativity between STEA and CTEA, between STEA and APL, between STEA and PCL, and also between APLP, APL and PCL, but there was no significant relativity between PCL and CTEA.

CONCLUSIONTKA for Chinese, the section of femoral posterior condyle should be external 5° to obtain the optimum rotational orientation. The property is different entirely between STEA and CTEA, the rotational alignment is not performed according to parallel to the CTEA in distal femur. Among STEA, APL, PCL, the STEA is the most reliable mark ofrotational alignment of the distal femur, and the PCL is the less reliable mark.

Adult ; Arthroplasty, Replacement, Knee ; Female ; Femur ; anatomy & histology ; diagnostic imaging ; Humans ; Male ; Middle Aged ; Rotation ; Tomography, X-Ray Computed

Adolescent ; Child ; Child, Preschool ; Cytomegalovirus Infections ; complications ; Female ; Humans ; Kidney Diseases ; etiology ; Kidney Glomerulus ; pathology ; Male

(1)H-NMR and (13)C-NMR assignments of 12-oleanene-3,11-dione (compound 1) were completely described for the first time through conventional 1D NMR and 2D shift-correlated NMR experiments using (1)H-(1)HCOSY, HMQC, HMBC techniques. Based on its NMR data, the assignments of 28-hydroxyolean-12-ene-3,11-dione (compound 2) were partially revised.
Euonymus ; metabolism ; Magnetic Resonance Spectroscopy ; Molecular Conformation ; Oleanolic Acid ; analogs & derivatives ; analysis ; chemistry ; Triterpenes ; analysis ; chemistry

Calcium-Binding Proteins ; metabolism ; Diagnosis, Differential ; Female ; Fibroma ; metabolism ; pathology ; surgery ; Humans ; Inclusion Bodies ; pathology ; Infant ; Microfilament Proteins ; metabolism ; Skin Neoplasms ; metabolism ; pathology ; surgery ; Soft Tissue Neoplasms ; pathology ; Tendons ; pathology ; Toes ; Vimentin ; metabolism

BACKGROUND: Recently biodegradable hydrogel has been extensively used to delivery anticancer drug and bioactive macromolecule. However, to protect the activity of the bioactive macromolecule, we need to obtain series of hydrogel which have milder hydrogelation conditions and shorter hydroglation time.OBJECTIVE: To prepare enantiomeric poly(L-Lactic acid) (PLLA)-poly(ethylene glycol (PEG)-PLLA/ poly(D-Lactic acid) (PDLA)-PEG-PDLA stereocomplex hydrogel which has shorter hydroglation time, to physically encapsulate a model drug-lysozyme and sustained release it from the hydrogel. METHODS: Triblock copolymers of PLLA-PEG-PLLA and PDLA-PEG-PDLA were synthesized by ring-opening polymerization of L(D)-lactide using PEG as the initiator and Sn(Oct)2 as the catalyst. The triblock copolymers were characterized by 1H nuclear magnetic resonance, FT-IR and X-Ray diffractometry. A hydrogel was prepared from an aqueous mixture of PLLA20-PEG227-PLLA20 and PDLA21-PEG227-PDLA21 (10 wt% concentration) at room temperature for 12 hours. X-Ray diffractometry test was used to research the gelation mechanism. The release profile of the lysozyme as a model drug from the hydrogel was tested. The morphology of the freeze-dried hydrogel was investigated by scanning electron microscope. The cytotoxicity of the hydrogel was evaluated by 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyl tetrazolium bromide) assay.RESULTS AND CONCLUSION: Triblock copolymers of PLLA-PEG-PLLA and PDLA-PEG-PDLA were obtained. Both the PEG and PLA blocks in the copolymers could crystallize, but the crystallization of the PEG block was predominant. The stereocomplex formation between the PLLA and PDLA blocks within the hydrogel was confirmed by the X-Ray diffractometry analysis. The release profile of the lysozyme from the hydrogel exhibited a sustained-release pattern with a duration period of 7 days. The hydrogel exhibited a 3D interconnected porous structure with 50-100 μm pore size after being freeze-dried. The mouse fibroblast cell viability percentage was 99.3% after the cells contacted with the 100% extracted liquid for 72 hours.

Objective To establish the ischemic precondition ([PC) model of PC12 cell line in vitro, and to explore the effect of nitric oxide (NO) on the IPC cerebral protection. Method PC12 cells were cultured and used for producing the model of ischemie precondition by the way of oxygen-glucose deprivation. Twenty dishes of cells were randomly divided into four groups (5 dishes for each group): control group, ischemic precondition group (IPC),non-ischemic precondition group (NIPC) and L-NAME treatment group (L-NAME). In control group, the cells were in-cubated with low glucose (<1 g/L) and2% FBS medium in normal oxygen; in IPC group, the cells were administrated with oxygen-glucose deprivation (OGD) for 6 hours, and then subjected with reperfuaion before OGD 15 hours; in NIPC group, the cells were treated the same as control group for 6 hours, and then subjected with reperfusion before OGD 15 hours; in L-NAME group, the cells received L-NAME (1 mmol/L) and cocultured for 30 minutes before OGD 6 hours, and then received the same treatment as the IPC group. To test whether the model was established, metabolic rate of MIT, LDH release were measured and the apoptosis rate was detected by flow cytometry following oxygen-glucose deprivation 15 hours. The activity of nitric oxide synthases (NOS) was as-sessed by biochemical assay. One-way ANOVA and LSD multiple comparison test were used to analyze differences among different groups, and P<0.05 was considered different. Results Compared with NIPC group, the metabolic rate of MTT increased (94.9%±35.1%, P<0.05), while LDH release and the cell apoptotic rate decreased significantly in IPC group (279.1%±28.1%, P<0.03). Compared with control group(100.0%± 13.5%),the activities of NOS increased both in NIPC and IPC groups (390.0%±14.6%, P<0.01;126.3% ±10.6%, P<0.01). Moreover, the apoptosis rates in each group (control group, IPC group, NIPC group and L-NAME group) were 5.90, 8.73, 38.62 and 11.73%,respectively. Conclusions IPC reduces the death and apoptosis rate of PC12 cell after oxygen-glucose deprivation injury. NO might be involved, but it is not the only factor.

Objective: To prepare a nanoprobe, anti-human melanoma ganglioside single chain variable fragment (GD/ScFvMEL) antibody conjugated with CdTe quantum dot, and to observe its ability to specifically bind human malignant melanoma cells. Methods: The GD/ScFvMEL gene was cloned into pET32a (+), and the plasmid was then transformed into E. coli BL21 (DE3) for GD/ScFvMEL protein antibody expression. The expressed GD/ScFvMEL antibody was purified by denaturing method and further refolded by modified dialysis method. The purified GD/ScFvMEL antibody was analyzed by SDS-PAGE. The GD/ScFvMEL-QDs nanoprobe was prepared by conjugating GD/ScFvMEL antibody with CdTe quantum dot, and its specificity was observed by incubating with MGC-803 cells and melanoma A375 cells. Results: The recombinant pET32a-GD/ScFvMEL was constructed and confirmed by PCR, restriction endonuclease analysis and DNA sequencing. The proportion of expressed GD/ScFvMEL antibody in total bacteria proteins was about 40% as detected by SDS-PAGE. The purified- and refolded-GD/ScFvMEL antibody was effectively conjugated with CdTe quantum dot, and the resulting GD/ScFvMEL-QDs nanoprobe was successfully prepared. The GD/ScFvMEL-QDs nanoprobe could specifically bind melanoma A375 cells, but could not bind stomach cancer MGC-803 cells. Conclusion: We have successfully prepared an anti-human melanoma ganglioside single-chain antibody-CdTe quantum dot nanoprobe, which can specifically bind melanoma cells.

Objective To investigate the effects of high spermatic vessel dissection on testicular morphological alteration of SD rats in prepuberty,puberty and sexual maturity phases.Methods Thirty-day-old SD rats were divided into 2 groups underwent sham operation and left high spermatic vessel dissection as a simulation of Palomo′s maneuver.Detailed morphological investigations were made at 3 different postoperative intervals among the 3rd day,30th day and 56th day.Results High spermatic vessel dissection in prepubertal rats induced acute testicular ischemia in the operated testes on the 3rd day.Most of the operated testes on the 30th day showed testicular atrophy.And all the operated testes showed testicular atrophy and sperm disappearance in epididymis on the 56th day.Conclusion High dissection of spermatic vessel in prepubertal rats induced testicular ischemia in prepuberty and testicular growth failure in puberty,testicular atrophy completely and sperm production losing in sexual maturity phase.