Background The visual evoked potential (VEP) is an important functional index to assess visual cortex plasticity.Monocular form deprivation(MD) of rat is a classic model to study visual cortex plasticity change.Utilizing VEP technique record the shift of ocular dominance in adult rats model is of an important significance for explore the treating opportunities of amblyopia.Objective The present study was to observe the pattern VEP changes in different ages of Long-Evens rats and the adult rats after binocular form deprivation.Methods Thirty-six healthy SPF Long-Evens rats were divided into PW3,PW4,PW5,PW6 groups according to the postnatal weeks,and PW7 rats included 36 SPF Long-Evens rats.The left eye lids of the rats were sutured in PW3,PW6 and PW7 rats for 3,5,7 days respectively,and PVEP in both left and right eye were recorded to assess the rat age that visual cortex plasticity ended.Bilateral PVEP were recorded in PW7 rats,and the bilateral eyelids were sutured to establish the bilateral form deprivation models.The right eyes were opened in 7,10,14 days,and the left eyes were opened in the next 3,5,7 days respectively and the PVEPs were recorded again to find the shift of ocular dominance and whether binocular form deprivation induce the visual cortex plasticity in adult rats.Results The P100amplitudes of the left eyes were gradually declined and those of the right eyes were raised from 3 to 7 days after MD of the left eyes in rats of PW3 group in comparison with before MD ( P＜0.01 ).In PW6 groups,no significant changes in P100amplitudes of both the right and left eyes were found in the third day after MD,but significant raise in the right eyes and lowing in the left eyes were seen in the 5 and 7 days after MD in comparison with before MD(P＜0.01 ).No any P100changes in both eyes were found in 3-7 days after MD compared with before MD in PW7 rats (P＞0.05).The C/I ratio (contralateral VEP amplitude to ipsilateral VEP amplitude in occluded eyes) in the rats of PW3,PW4 and PW5 groups were 1.07±0.15,1.16±0.16 and 1.14±0.15 respectively in 3 days after MD,showing the significant lowing in comparison with before MD (2.69±0.45,2.58±0.4,2.62±0.32) (P＜0.01),but those of PW6 and PW7 were unchanged (2.80±0.48 vs 2.90±0.46,2.59 ±0.36 vs 2.90±0.46,P＞0.05 ),indicating the absence of ocular dominance plasticity in the adult rats.In rats of PW7 with binocular form deprivation for 14 days,a significant decrease was observed in the C/I ratio in rats with next MD for 3 days,demonstrating that the visual cortex plasticity was reactivated after 14 days of binocular form deprivation in PW7 rats( 1.33±0.18 vs 2.70±0.45,P＜0.01 ).Conclusions PVEP can be recorded in Long-Evens rats.It is a major index for identifying the shift of ocular dominance in the Long-Evens rats.Binocular form deprivation can reflect the visual cortex plasticity in the adult rats.

10.3969/j.issn.1007-3969.2013.05.004

BACKGROUND:Distal femur is a predilection site for primary bone tumors, most of which are treated with salvage treatment. There are many reports on the early-stage, mid-term and prognosis of the distal femur tumor-type artificial knee joint prosthesis, but reports on long-term function and prognosis are less.
OBJECTIVE:To study the long-term prognosis and limb function after treatment with rotating-hinge knee prosthesis.
METHODS:Thirty-four patients with distal femoral bone tumors admitted to the Department of Orthopedics, the First Affiliated Hospital of Anhui Medical University between January 2000 and June 2008 were retrospectively analyzed, including 19 males and 15 females. Al patients were subjected to rotating hinge knee prosthesis replacement.
RESULTS AND CONCLUSION:The mean fol ow-up period was 62.4 months (range 11 to 126 months). At the final of fol ow-up, 23 patients were event-free alive and 11 patients were dead. Seven of 23 survival patients suffered from prosthesis related complications, including four cases of prosthesis revision and three cases of amputation. The event-free survival rate of 23 patients undergoing replacement of domestic tumor prosthesis around the knee was 67.6%in 5 years. The mean functional score of affected limb was 19.30 (7 to 27 points):excellent in 6 cases, good in 16, fine in 5 and poor in 2. The excellent-good rate was 75.9%. The clinical outcomes suggested that the satisfactory limb function and survival rate could be achieved with the application of domestic tumor prosthesis replacement for treatment of malignant bone tumors around the knee.

Objective Increased renal mast cells have been found in significant correlation with clinicopathological features of diabetic nephropathy ( DN) .However, the exact pathological significance of mast cells still remained to be elucidated.As one of the most abundant serine proteases, tryptase is specifically expressed by mast cells.The study was to observe the expression of tryptase in the urine of patients with diabetic nephropathy and realize the pathological significance of urinary tryptase and renal mast cells in the development of diabetic nephropathy. Methods Urine samples from 103 patients with diabetic nephropathy who were hospitalized at the clinical unit of the nephrology centre of Jingling Hospital from January 2012 to June 2013 were collected.According to concentra-tion of urinary albumin excretion rate, the patients were conducted to early-stage DN group (microalbuminuria,n=10), middle-stage DN group (proteinuria stage, n=31) and end-stage DN group (renal insufficiency stage,n=62).Meanwhile, urine samples from 20 normal persons were taken as the normal control group.Tryptase level in each urine sample was measured by using an ELISA kit.The average tryptase levels were compared among different groups and the correlation between urinary tryptase level and clinical indices was analyzed. Results ①In most cases, tryptase was not detected in the urine samples from normal persons(0.7 ±1.4 ng/mg creati-nine).However, the urinary trypatse level increased significantly with the development of diabetic nephropathy (11.6 ±10.5 ng/mg creatinine for early stage, 29.7 ±19.2 ng/mg creatinine for middle stage, and 44.6 ±43.4 ng/mg creatinine for late stage group).②The urinary tryptase level was found in significant correlation with ser-um creatinine (r=0.325, P<0.01), serum cystatin C (r=0.391, P<0.01), serum urea nitrogen (r=0.27, P<0.01), 24 hour uri-nary protein (r=0.389, P<0.01), urine C3 (r=0.276, P<0.05), urine retinol-binding protein (r=0.365, P<0.01), urine lysozyme (r=0.461, P<0.01), urine N-acetyl-β-glucosaminidase level (r=0.568, P<0.01), urinary kidney injury molecule-1 (r=0.434, P<0.05) and interleukin-18 level (r=0.375, P<0.05).No significant correlation was found between urinary tryptase level and age, gender, fasting blood sugar, postprandial blood sugar, HbA1c, triglyceride, high density lipoprotein or low density lipo-protein. Conclusion Mast cells play an important role in the development of diabetic nephropathy and might be a novel and effective target for the treatment of DN.

Objectives To explore the relationship between serum leve ls of hepatocyte growth factor (HGF) and matrix metalloproteinase-9 (MMP-9) and the disease activity of systemic lupus erythematosus (SLE), and study the mechan isms of these two factors in the pathogenesis of SLE. Methods The serum levels of HGF and MMP-9 were measured by ELISA. The growth curve of normal ECV304 cell line was obtained, and the action concentration of recombinant human hepatocyte growth factor (rhHGF) was determined. MMP-9 expression level in cells was detec ted by flow cytometric analysis. Results The serum level of HGF increased sign ificantly in SLE patients as compared with that in healthy controls (P 0.05). The area of ROC curve was 0.707 and the sensitivity was 66 .7% when using the serum level of HGF as diagnostic standard. The area under ROC curve was 0.984 and the sensitivity was 97.2% when using the serum level of MMP-9 as diagnostic standard. The sensitivity was 66.7% (24/36) when two markers (H GF and MMP-9) were examined simultaneously. Additionally, the action concentrati on of rhHGF was 8 ng/mL, and the expression level of MMP-9 was 39.74% in normal ECV304 cells and increased to 40.32% after rhHGF stimulation. Conclusions It i s suggested that HGF and MMP-9 may be involved in the pathogenesis of SLE, and s erum levels of HGF and MMP-9 might be used as markers for monitoring the disease activity, renal damage, disease progression and improvement in SLE. The sensiti vity might be higher when serum level of MMP-9 is used as diagnostic standard, a nd rhHGF can enhance MMP-9 expression in ECV304 cell line.

Objective To investigate the relationship between serum level of MMP-9(matrix metalloproteinase-9) and the disease activity of systemic lupus erythematosus(SLE).Methods Serum level of MMP-9 of 30 controls and 36 SLE patients were measured by Enzyme Linked Immuno Sorbent Assay.Results Significantly decreased serum level of MMP-9 was found in SLE patients as compared to that in healthy controls(108?113) ng/ml vs (352?155) ng/ml (P0.05).Conclusion The present data suggests that MMP-9 may be involved in the pathogenesis of SLE,and serum MMP-9 level may be a marker of disease activity,renal damage,disease progression and amelioration in SLE.

Objective Our previous study showed that the expression of C 3aR was increased in renal tubular epithelial cells (RTEC).However, the role of C3aR in diabetic nephropathy remains unknown, and so does the exact physiological and pathological signifi-cance of C3aR in relevant renal tissues.In the present study, we investigated the physiological and pathological significance of C3aR in RTECs using a lentivirus expression vector and anRTEC strain overexpressing C3aR. Methods Based on the sequence of C3aR mRNA, the human C3aR gene was synthesized and cloned into the multi-clonal site of the lentivirus expression vector pLenti6.3-MCS-IRES2-EGFP to construct a C3aR expression vector pLenti6.3-C3aR-IRES2-EGFP.After identification by sequencing, the recombinant lentivirus expressing C3aR was packaged by cotransfecting293T cellswith the recombinant lentivirusexpression vector and packaged plasmid .Then,the recombinant lentivirus was used to infect the human RTECline HK 2.After screening in the medium with blasticidin, blasticidinr-esistant cell clones were obtained, followed by identification ofthe human RTECline stably overexpressing C 3aR by real-time PCR and immunochemical staining. Results TheC3aR expression vector pLenti6.3-C3aR-IRES2-EGFP was successfully constructed and the sequence was proved to be correct. C3aR expression recombinant lentivirus was successfully packaged with a titer of 5×108/mL.Blasticidin-resistant HK2 cell clones were ob-tained and the expression of HK2-C3aR mRNA was significantly higherin the HK2-C3aR cells than in the non-transfected HK2 cells (2.33± 0.45 sv 1.00±0.09, P<0.05). Conclusion We successfully constructed a C3aR expression lentivirus vector pLenti6.3-C3aR-IRES 2-EGFP and a C3aR overexpression renal tubular epithelial cell strain HK2-C3aR, which is very contributive to further studies of the roles of C3aR in renal tubular epithelial cells and other types of cells.

Objective To investigate the differences in sex hormone levels between hypospadias and circumcision groups.Methods Fifty cases of circumcision and 137 cases of hypospadias the dihydmtestosterone (DHT) value was tested with radioimmunoassay,and testosterone was tested with lightimmunoassay.Results DHT value was (64.51 ±32.10)pg/ml in circumcision group,and (46.72 ±28.94)pg/ml in hypospadias group (P ＜0.05).DHT value in hypospadias type Ⅰ,Ⅱ,Ⅲ,and Ⅳ were (50.20 ±32.90)pg/ml,(46.63 ±25.67)pg/rnl,(51.60 ±32.16)pg/ral,and (39.02 ±26.32)pg/ml,respectively (P =0.29).The differences between circumcision and hypospadias groups were statistically significant (P =0.00).Luteinizing hormone (LH) and testosterone (T) in children with hypospadias were significantly lower than those in children in the circumcision group (P ＜ 0.05).No statistically significant difference was found between two groups in follicle-stimulating hormone (FSH),estradiol (F2),and prolactin (PRL) (P ＞ 0.05).No statistically significant differences were found in FSH among all types of hypospadias (P ＞ 0.05).Conclusions Inadequate secretion of T or activity insufficiency and functional deficiency of 5 alpha reductase (SRD5A) are likely found in children with hypospadia.Inadequate secretion and low T value might be found in LH-T shaft in children with hypospadia.The normal T value in some children with hypospadia does not show that androgens produced during pregnancy are normal.

Objectives To explore the perceived stigma among family members of people with schizophrenia and to identify the influential factors related to perceived stigma in socio-demographic characters of family members and patient characters.Methods Total 127 family members of people with schizophrenia in a psychiatric hospital in Beijing were surveyed by the Perceived Devaluation-Discrimination Scale developed by Link.Results The mean score of the perceived devaluation-discrimination scale is 2.60±0.44.It was significantly higher than the 2.5 midpoint(t=2.53,P<0.05).Family member's perceived stigma was significantly different between different groups of residence area,education level,occupation and family income (P<0.05).Perceived stigma were not affected by patient characters (P>0.05).Family members'education level was the influencing factors of stigma,and the R~2 Square is 0.213.Conclusions The family members of people with schizophrenia suffered from high stigma.Education level of family member may be one of the influential factors.

Objective To investigate the effect of histone deacetylase inhibitor on Her2 positive breast cancer cell line BT474 and SKBR3 in apoptosis and metastasis.Methods Histone deacetylase inhibitor MS-275,suberoylanilide hydroxamic acid (SAHA)(4 μmol/L,and 50 μmol/iL,respectively) treated the cell lines BT474 and SKBR3 cells.Flow cytometer examined the apoptosis ratio.Transwell tested their metastatic activity.Western blot assay was performed to detect the associated proteins.Results SAHA and MS-275 inhibited the cell survival.The BT474 cell survival was (39 ± 11) ％,(54 ± 8) ％,and the SKBR3 survival was (62 ± 6) ％,(71 ± 9) ％,according to the fluorescence-activated cell sorting (FACS) result.SAHA and MS-275 induced the BT474 cell apoptosis 8.46± 0.28 (P ＜0.01),4.15 ± 0.71 (P ＜0.01) fold change,respectively;and upregulated the SKBR3 cell apoptosis ratio 5.51 ± 1.24 (P ＜0.01),4.04 ±0.69 (P ＜0.01) fold.The Transwell result showed that SAHA,MS-275 inhibited the Transwell ability of BT474 from the control 184.7 ± 18.8 to 104.3 ± 7.1,131.3 ±9.1 per view,and the SKBR3 from control 60 ± 16.7 per view to 14.3 ± 6.5,34.3 ± 8.7 per view.The Western blot result showed that SAHA,MS-275 inhibited the protein level of vimentin,Her2,β-catenin,histone deacetylase inhibitor (HDACi),and upregulated the acetylation level of histone 3.The E-caherin protein was regulated in BT474 and SKBR3 cells.Conclusions MS-275,SAHA can induce BT474 and SKBR3 apoptosis significantly,also inhibit their metastatic activity.