Objective To study the relationship between urinary laminin(LN) and diabetic nephropathy.Methods Using the method for concentrating urinary laminin based on precipitation with PEG-4 000 and γ-globulin.The concentration of LN in the urine of 83 diabetics was measured by RIA.The concentration of micro-albumin and Ccr in the urine and the levels of plasma glucose and HbA1c were also measured.Results The choice of 150g/L of PEG-4 000 and 0.5 g/L of γ-globulin was the best method for concentrating urinary LN.The urinary LN in all groups was significantly higher than that in control group.Conclusion Urinary LN is an indicator in the diagnosis of the early diabetic nephropathy.

BACKGROUND: Compared to those original viruses systems, adeasy adenovirus, a recombinant adenoviral system widely used in recent years, based on viruses with a deletion of both El and E3, reported by T.C. He in 1998, is an improved one. It simplifies the generation and production of such viruses and expedite the process of generating and testing recombinant adenoviruses using homologous recombination in bacteria rather than in eukaryotic cells. Moreover, it can be conveniently followed with the aid of green fluorescent protein encoded by EGFP gene incorporated into the viral backbone.OBJECTIVE: To construct the recombinant adenovirus and to evaluate them by transfect them to mesenchymal stem cells (MSCs)and detect the expression of target gene hlGF-I at gene and protein levels.DESIGN: Repetitive measurement wail.SETTING: The Institute of Pediatric Research, Chongqing University of Medical Science.METHODS: The study was performed at the Institute of Pediatric Research, Chongqing University of Medical Science from November 2004 to March 2005. After the amplification of truncated hlGF-1 gene from pcDNA3.l-hlGF-I by polymerase chain reaction (PCR), the gene fragment was inserted into the shuttle plasmid pAdtrack-CMV for homologous recombination with backbone plasmid pAdeasy-I in bacteria BJ5183 to get adenovirus.Ad-hlGF-1. The high titer adenovirus supernatant was obtained by repeated transducing of HEK 293 cells by adenovirus harvested after confirmation of the adenovirus structure. As target cells,MSCs were infected with adenovirus earned target gene, hIGF-1, to determine the expression of hlGF-1 gene.MAIN OUTCOME MEASURES: ① The construction of recombinant adenovirus vector;② the expression of target gene hIGF-1 in HEK 293 cells and the proper multiplicity of infection (MOI); ③ hIGF-1 gene expression in MSCs.RESULTS: The adenovirus vector based on adeasy system was constructed successfully and the Ad-hlGF transducing was successfully or efficiently expressed in MSCs cells. The ideal expression of harvested recombinant adenovirus in MSCs was detected by fluorescence microscope, RT-PCR, immunocytochemistry, and Western Blot.CONCLUSION: Adenovirus vector is an effective vector tools for gene expression and wansfection of MSCs. MSCs transduced with Ad-hIGF-1 maybe another option to gene-modified seed cells for articular cartilage tissue engineering.

ObjectiveTo investigate the clinical significance of posterior spinal cord shifting after two different types of laminoplasty for multilevel cervical myelopathy.Methods From June 2004 to September 2007,65 patients with cervical spondylotic myelopathy were reviewed in this study,including 41 males and 24 females with an average age was 56.3 years(range,39-75).Among them,33 patients underwent selective laminoplasty and 32 patients underwent open-door laminoplasty.There were no significant differences of preoperative JOA scores and cervical curvature index between two groups.After one year follow -up,the posterior shifting of spinal cord,Japanese Orthopaedics Association (JOA) recovery rate,loss of curvature index and axial symptom for each patient were calculated.ResultsAll cases were followed up for average 34 months(range,26-47).There was significant difference of the postoperative posterior shifting between the two groups,which was(1.4±0.6) mm in selective laminoplasty group and(3.3±1.2) mm in opendoor laminoplasty group,respectively.The average JOA recovery rate was 60.5％±21.3％ and 61.1％±17.9％ in selective laminoplasty and open-door laminoplasty group,respectively.There was no significant difference of JOA recovery rate between the two groups.Loss of cervical curvature indices was 3.3％±1.7％ and 3.1％± 2.4％ in selective laminoplasty and open-door laminoplasty group,respectively,with no significant difference between the two groups.The rate of patients with evident axial symptoms was 18.2％ and 33.3％ in selective laminoplasty group and open-door laminoplasty group,and the difference of the scores of cervical axial symptom was statistically significant.ConclusionThe degree of the postoperative posterior shifting of the spinal cord in open-door laminoplasty group was greater than that in selective laminoplasty group.The posterior shifting of the spinal is correlated with cord,axial symptom but not the recovery rate and curvature index.

BACKGROUND:Previous studies have found that miR-21 expression is increased during osteogenic differentiation of bone marrow mesenchymal stem cells, but the action and molecular mechanism of miR-21 are stil unclear. OBJECTIVE:To verify the target gene of miR-21, Spry1, and to explore the role of Spry1 in osteogenic differentiation of human bone marrow mesenchymal stem cells. METHODS:Luciferase report was used to verify Spry1 gene targeted by miR-21, and western blot assay was used to detect the expression of Spry1 in the osteogenesis of human bone marrow mesenchymal stem cells. Spry1 expression vector was established and transfected into human bone marrow mesenchymal stem cells. Osteogenesis ability of human bone marrow mesenchymal stem cells was analyzed after Spry1 high expression by alkaline phosphatase, alizarin red staining, RT-PCR and western blot. RESULTS AND CONCLUSION:Luciferase report suggested that Spry1 was a target gene of miR-21. The expression level of Spry1 was decreased in the osteogenesis of human bone marrow mesenchymal stem cells. Increasing expression of Spry1 could inhibit osteogenic differentiation of human bone marrow mesenchymal stem cells. These results indicate that Spry1 as a target gene of miR-21 negatively regulates osteogenic differentiation of human bone marrow mesenchymal stem cells, and plays an important role in bone formation process.

Objective To investigate the situation of food contamination by lead,cadmium,mercury,copper and aluminum in Ningxia Region.Methods Ten categories of food samples up to 712 copies including dairy,edible mushrooms,eggs,preserved egg,meat,organs,aquatic products,vegetables,grains and pasta from supermarkets of five administrative cities of Ningxia Autonomous Region were selected in 2010.The contents of lead,cadmium and mercury in food were detected in 646 samples of 9 categories of food,and copper content of the 30 meat samples were detected.Aluminum content in 66 samples of flour food was tested.According to the methods of national food safety standards (GB 5009.12-2010) and national food hygiene standards (GB/T 5009.13-2003,GB/ T 5009.15-2003 and GB/T 5009.17-2003),lead,copper,cadmium and mercury in various foods were tested.Aluminum was determined using the method for determination of spectrophotometry.Results The detection rates of lead,cadmium,mercury in the nine categories of food were 52.8％ (341/646),62.1％ (401/646),51.7％ (334/646);the detection rates of copper in the meat was 90.0％ (27/30);the aluminum in flour food was 84.8％ (56/66),respectively.Compared with the national standards,the eligible rates of lead,cadmium,mercury,copper and aluminum were 97.1％ (627/646),98.2％ (539/549),100.0％ (646/646),100.0％ (30/30) and 75.8％ (50/66),respectively.Food contaminated by lead and cadmium were mainly distributed in preserved egg [lead detection rate 9.5％ (2/21)],organs [lead detection rate 6.7％ (2/30),cadmium detection rate 30.0％ (9/30)],vegetables [lead detection rate 9.9％ (8/90)] and a small amount of food products [lead detection rate 4.7％ (7/150)].The levels of lead in fried flour food and fine dried noodles were higher [40.9％ (9/22),36.4％ (4/11)].Conclusions Food contamination by lead,cadmium,mercury and copper is not serious in Ningxia.The monitoring results of lead,cadmium,mercury and copper in food are satisfying in Ningxia.But there is aluminum contamination in flour food.It is important to do the risk evaluation of aluminum contamination in food as quickly as possible.

OBJECTIVE:To provide reference for rational use of anti-tumor essential medicine in the clinic. METHODS:The application of anti-tumor essential medicine in 39 hospitals from Chongqing area during 2012-2014 was analyzed statistically in re-spects of consumption sum,DDDs,DDC and B/A. RESULTS:The total consumption sum of anti-tumor essential medicine in 39 hospitals from Chongqing area during 2012-2014 increased from 10 083.65 ten thousand yuan to 15 186.65 ten thousand yuan,with annual increase rate of 22.72%. 3-year total consumption sum was 37 952.45 ten thousand yuan,and the sum of consumption sum of top 10 medicines was 36 742.24 ten thousand yuan,accounting for 96.81%;top 3 medicines were paclitaxel,oxaliplatin and flu-orouracil. During 2012-2014,DDDs had changed to certain extent,and the front medicines contained tegafur,paclitaxel and cyclo-phosphamide,etc;the most increment medicine was paclitaxel (oral),whose annual growth rate was 59.63%;that of tamoxifen decreased year by year,with annual decrease rate of 5.14%. DDC of paclitaxel,oxaliplatin and etoposide were the higher others. Annual B/A of oxaliplatin,fluorouracil and cytosine arabinoside were all lower than 1.00. CONCLUSIONS:The lonsumption sum and DDDs of anti-tumor essential medicine had increased reasonably in 39 hospitals from Chongqing area during 2012-2014,while their DDC had kept stable. Main medicines used in the clinic are suitable for numerous indications,and are expensive.

BACKGROUND:Osteogenic differentiation is a complex process involving transcriptional and post-transcriptional regulation by multiple signaling pathways, and the specific mechanisms remain unclear. It is of great significance to study the role of critical miRNAs in the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in the treatment of osteoporosis and bone defects. OBJECTIVE: To explore the regulatory ability of miR-21/Sprouty1 function axis in the osteogenic differentiation of BMSCs in patients with postmenopausal osteoporosis. METHODS:BMSCs were isolated from healthy people (H-hBMSCs) and patients with postmenopausal osteoporosis (PMOP-hBMSCs), and their osteogenic ability was compared. Expression of miR-21 and Spry1 at gene and protein levels was detected by real-time RT-PCR and western blot assay, respectively. miR-21 expression was upregulated via transfection in PMOP-hBMSCs, and the osteogenic ability and Spry1 expression of the cells were detected, while real-time RT-PCR and western blot were used to detect the expression of osteogenic marker genes, Runx2 and Osterix. RESULTS AND CONCLUSION:Compared with H-hBMSCs, PMOP-hBMSCs osteogenic ability was weakened significantly, miR-21 expression decreased, and Spry1 expression increased, indicating an inhibition to the miR-21-Spry1 function axis. Through the transfection of miR-21 and down-regulation of Spry1, the expression levels of Runx2 and Osterix were increased, and PMOP-hBMSCs osteogenic ability was partially restored.

Objective:To explore the predictive value of computed tomography (CT) texture analysis for the recurrence in children with acute pancreatitis (AP).Methods:The clinical diagnostic test was conducted. The clinical data of 56 children with primary AP who were admitted to Wuhan Fourth Hospital from January 2016 to January 2018 were collected. There were 13 males and 43 females, aged from 3.5 to 13.0 years, with a median age of 5.5 years. Based on follow-up in other hospitals, 20 children with recurrence of AP were allocated into recurrence group, and 36 children without recurrence were allocated into non-recurrence group. All the 56 children underwent abdomen plain and enhanced CT scan within 24 hours after first admission. Observation indicators: (1) comparison of clinicopathological features between two groups of children with AP. (2) comparison of CT texture parameters between two groups of children with AP. (3) diagnostic efficacy of clinical features and CT texture parameters. Follow-up using outpatient reexamination and telephone interview was conducted to detect recurrence of AP up to February 2020. The duration of follow-up required more than or equal to 24 months. The Shapiro Wilk test was used to analyze normality of measurement data. Measurement data with normal distribution were repre-sented as Mean± SD, and comparison between groups was conducted using the t test. Measurement data with skewed distribution were represented as M (range) or M ( P25, P75), and comparison between groups was conducted using the Mann-Whitney U test. Count data were described as absolute numbers, and comparison between groups was analyzed using the chi-square test. Comparison of ordinal data was conducted using the Mann-Whitney U test. Clinical parameters and CT texture parameters with statistical differences were multivariate analyzed using the Logistic regression model. Receiver operating characteristic curve (ROC) analysis was used to evaluate the predictive efficacy of parameters for recurrence of AP. Results:(1) Comparison of clinicopatholo-gical features between two groups of children with AP: cases with or without complications were 14 and 6 for the recurrence group, versus 7 and 29 for the non-recurrence group, showing a significant difference between the two groups ( χ2=14.021, P<0.05). Cases with minimal, moderately severe or severe disease (severity of disease) were 2, 5, 13 for the recurrence group, versus 19, 11, 6 for the non-recurrence group, showing a significant difference between the two groups ( Z=5.414, P<0.05). (2) Comparison of CT texture parameters between two groups of children with AP: the energy value in the arterial phase on CT examination was 0.186(0.174,0.206)for the recurrence group and 0.413(0.405,0.425) for the non-recurrence group, showing a significant difference between the two groups ( Z=9.413, P<0.05). The energy value and entropy value in the venous phase on CT examination were 0.084(0.078,0.092) and 0.961(0.210,1.720) for the recurrence group, versus 0.135(0.124,0.156) and 0.372(0.210,0.535) for the non-recurrence group, showing significant differences between the two groups ( Z=4.763, 7.243, P<0.05). (3) Diagnostic efficacy of clinical parameters and CT texture parameters: results of multivariate analysis showed the complications, severity of disease, energy value in the arterial phase on CT examination were related factors for recurrence in children with AP, energy value and entropy value in the venous phase on CT examination were related factor for recurrence in children with AP ( odds ratio=0.874, 0.765, 0.837, 0.902, 0.813, 95% confidence interval as 0.802?0.985, 0.581?0.914, 0.753?0.897, 0.862?0.948, 0.765?0.873, P<0.05). Results of ROC analysis showed that that areas under curve (AUC) of complications, severity of disease in the clinical parameters were 0.734 and 0.832, the AUC of single CT texture parameter was 0.811?0.867, the AUC of clinico-pathological parameters combined with CT texture parameters was 0.882. Conclusion:CT texture analysis can early and non-invasively predict the recurrence of AP in children, and the combination of clinicopathological parameters with CT texture parameter has a better predictive efficacy.

Objective To evaluate the clinical value of 18F-FDG PET-CT in monitoring response to therapy in patients with follicular lymphoma (FL) staging Ⅲ-Ⅳ.Methods The clinical data of 20 patients with FL were analyzed retrospectively.All patients underwent PET-CT at three times:baseline,after 3-4 cycles of chemotherapy,and end of treatment.Kaplan-Meier method was used to analyze the survival of patients.Results 14 patients achieved CR at the interim PET-CT,all patients achieved CR at post therapy PET-CT:all patients were alive with no evidence of disease in the follow-up.2 patiens achieved PR at the interim PET-CT,and 1 patient achieved CR,and 1 patient was PD,at post therapy PET-CT,all patients were relapsed in the follow-up.4 patients achieved PD,and all patients were changed the chemotherapy plan.At post therapy PET-CT,2 patients achieved CR,1 patient achieved PR,and 1 patient was PD,1 patient was no evidence of disease,2 patients were relapsed,and another was died in the follow-up.The negative predictive values (NPV) of the interim and the post therapy PET-CT were 100 ％ (14/14),88.2 ％ (15/17) respectively.The interim PET-CT was no significantly correlated with PFS (P =0.135),while the post therapy PET-CT was significantly correlated with PFS (P =0.0006).Conclusion 18F-FDG PET-CT is useful to monitor response to chemotherapy in patients with FL staging Ⅲ-Ⅳ,and set personalized treatment plan.

Objective Subarachnoid hemorrhage ( SAH) is a devastating disease with high fatality and morbidity and micro-glia/macrophage ( M/M) plays a vital role in SAH brain injury with complicated pathophysiological mechanism .This study was to ob-serve the effect of Nrf2 deficiency on M/M activation and M1 polarization after subarachnoid hemorrhage in mice . Meth ods We col-lected 70 wild-type ( WT) ICR mice and 35 Nrf2-knockout ( KO) mice to establish the SAH model by injecting fresh autologous blood into pre-chiasmatic cistern.WT mice were arranged into four groups: sham operation group, post operative day 1 (POD1) group, POD3 group and POD5 group.Then WT mice and Nrf2 Nrf2-knockout mice were divided into sham operation WT group , sham opera-tion KO group, SAH WT group and SAH KO group.Western blotting (WB) and immunohistochemistry (IHC) were applied to observe the activation and proliferation of M/M after SAH on WT mice .Difference in activation and M 1 polarization were observed by detecting Iba1 expression in WB and CD 16/32 +Iba1 +cells in immunofluorescence between WT and KO mice . Results Gray scale values of Iba1 expression by WB in WT mice are 0.491 ±0.039, 0.657 ± 0.069, 0.930 ±0.046 and 0.926 ±0.046;average optical intensity values of Iba1 expression by IHC in WT mice are 0.412 ±0.122, 0.625 ±0.135, 0.963 ±0.213 and 0.978 ±0.224.The data indica-ted that Iba1 expression increased in SAH KO group in comparison to SAH WT group on 1, 3, 5 day after SAH (P<0.05).Moreover, Nrf2 deficiency promoted the activation and polarization of M /M by increased Iba1 protein expression and CD16/32 +Iba1 +cells after SAH ( P<0.05). Conclusion SAH induces M/M activation and proliferation in mice, and Nrf2 deficiency promotes the activa-tion, proliferation and M1 polarization after SAH .