Objective To investigate the application of PKH26 and molecular light imaging system in cartilage en?gineering. Methods Canine chondrocyte was labeled by fluorescent dye PKH26 and seeded into the porous cartilage acel?lular matrix scaffold. The cells/scaffold constructs were cultured in vitro for 1 week. Then the constructs were implanted into the dorsal pocket of nude mice. We utilized a molecular light imaging system to macroscopically observe cells/scaffold con?structs in vivo with fluorescence at the 4th weeks, and compared with X-rays taken at the same position. The fluorescence im?ages were compared with the immunohistochemical and immunofluorescent results of cartilage-like tissue in vivo. Results Luminescent images were acquired at the 4th weeks, a red color enhanced overlay of the luminescent image over X-ray photo?graphic image demonstrated the location of the implants and the cell viability and cell growth on porous CACM scaffold in vivo were very well. Histological results show that the safranin O, anti-collagenⅡimmunohistochemistry and toluidine blue stain of cartilage-like tissue is positive. Immunofluorescence examination demonstrated chondrocytes in the constructs whitch is showen red fluorescence, and anti-collagenⅡimmunofluorescent staining was showen in green while the overlap?ping image is showen in yellow. Conclusion This study outlines an applicable non-destructive method to evaluate cell growth in tissue engineering constructs in vivo using PKH26 and molecular light imaging system.

Increasing the production and secretion of endogenous opioid peptide by immune cell can significantly induce myocardial protective effects against ischemia-reperfusion injury. Gene therapy is promising to increase endogenous enkephalin (ENK). However, classical viral and plasmid vectors for gene delivery are hampered by immunogenicity, gene recombination, oncogene activation, the production of antibacterial antibody and changes in physiological gene expression. Minimalistic immunologically defined gene expression (MIDGE) can overcome all the deficients of viral and plasmid vectors. The exon of rat's preproenkephalin (PPENK) gene was amplified by PCR and the fragments were cloned into pEGFP-N1 plasmids. The recombined plasmids were digested with enzymes to obtain a linear vector contained promoter, preproenkephalin gene, RNA stable sequences and oligodesoxy nucleotides (ODNs) added to both ends of the gene vector to protect gene vector from exonuclease degradation. A nuclear localization sequence (NLS) was attached to an ODN to ensure the effective transport to the nucleus and transgene expression. Flow cytometry, laser confocal microscopy and Western blotting demonstrated that PPENK-MIDGE-NLS can transfect leukocyte of rat in vivo, increase the expression of proenkephalin (PENK) in tissue, and the transfection efficiency depends on gene vector's dosage. These results indicate that PPENK-MIDGE-NLS could be an innovative method to protect and treatment of myocardial ischemia-reperfusion injury.
Animals ; Cloning, Molecular ; Enkephalins ; genetics ; Gene Expression ; Genetic Therapy ; Genetic Vectors ; Leukocytes ; Plasmids ; Promoter Regions, Genetic ; Protein Precursors ; genetics ; Rats ; Transfection ; Transgenes

BACKGROUND: The dynamic characteristics of electroencephalogram (EEG) include a decrease in the chaotic dimension, the correlation dimen sion, the Lyapunov exponent, the chaotic complexity, the freedom of EEG and an enhanced synchronization and periodicity of the EEG from several minutes to tens of minutes before epileptic seizures. All these characteristics prefigure the forthcoming seizures. Some studies have proven that the non linear dynamical system can be used as a feasible approach to explore the potential variables for describing the chaos portrait of EEG. OBJECTIVE: To analyze the electric characteristics of EEG signal in the epileptic seizures in rat model by investigating the nonlinear dynamical variables, such as the approximate entropy (ApEn) and correlation dimen sion. DESIGN: Observational and experimental study based on animals. SETTING: Department of Medical Engineering, Department of Gastroen terology, Second Artilleryman General Hospital of Chinese PLA; Department of Physics, Faulty of Biomedical Engineering, Fourth Military Medical Uni versity of Chinese PLA. MATERIALS: From September 2001 to January 2002, this study was conducted at the Complexity Laboratory of the Biomedical Department of the Fourth Military Medical University of Chinese PLA. Six male SD rats,weighing 150- 200 g, were selected.INTERVENTIONS: After intraperitoneal injection of chloral hydrate (0. 5 mL), the male SD rats were deeply anesthetized. When their EEG signal became stable, bemegride injection was diluted at 1:1 with saline and was given on a volume of 0.5 mL to the rats intraperitoneally. After a while,the epileptic seizures started marked by a spasm with a deep roar. The entire epileptic seizures were recorded. According to the shape of EEG waves and the corresponding symptoms of the rats during their seizures, data of the four phases, referring to normal condition, preictal phase, ictal phase and postictal phases of epileptic seizures, were selected for nonlinear analysis. The variations of the ApEn and the correlation dimension were calculated.MAIN OUTCOME MEASURES: In the four phases of the seizures, before seizures, preictal phase, ictal phase and postictal phases, the changes in the ApEn and correlation dimension were observed.RESULTS: All the 6 rats entered the statistical procedure. During epilepsy, the ApEn and correlation dimension of the EEG signal in ictal phases (0. 447 ±0. 126, 2. 166 ±0. 377) decreased significantly while those in preictal phases(0. 807 ±0. 182, 4. 773 ±0. 319) and postictal phases (1. 241 ±0. 125, 6. 042 ±0. 373) (t = -3. 984to 17. 902, P ＜0. 01). The ApEn and the correlation dimension of the EEG signal in preictal and ictal phases had significant difference with those observed under normal conditions (1.313 ± 0. 090, 6. 405 ± 0. 694) (t = -5. 228 to 12. 740, P ＜ 0. 01 ).CONCLUSION: The changes in ApEn and correlation dimension showed by nonlinear dynamical approach in this study reflect the characteristics of EEG signals in preictal time, ictal time and postictal timeof the epileptic seizures and the differences among them. Additionally, they also reveal the laws in the changes of the complex ictal EEG signal.

Objective:To investigate the clinicopathological characteristics and influencing factors of kidney prognosis in primary IgA nephropathy (IgAN) patients.Methods:The data of primary IgAN patients diagnosed with renal biopsy in the First Affiliated Hospital of Anhui Medical University from January 2015 to September 2019 were retrospective analyzed. According to the level of baseline estimated glomerular filtration rate (eGFR) when performing renal biopsy, the patients were divided into group A[eGFR≥90 ml·min -1·(1.73 m 2) -1], group B[eGFR 61-89 ml·min -1·(1.73 m 2) -1] and group C[eGFR≤60 ml·min -1·(1.73 m 2) -1]. The clinical and pathological data were collected and compared among the three groups. Kaplan-Meier method was conducted for renal results, whereas the Cox proportional-hazards regression model was exploited to analyze the influencing factors of kidney prognosis in IgAN patients. Results:A total of 742 patients were included in the study, including 394 cases (53.1%) in group A, 203 cases (27.4%) in group B, and 145 cases (19.5%) in group C. There were 325 males (43.8%) and 417 females (56.2%). The median duration of renal biopsy was 6 (1, 24) months, and the median age was 36 years old (18-68 years old). As the baseline level of renal function decreased, the proportion of patients with nephrotic syndrome, hypertension, anemia and hyperuricemia and the levels of 24 h urinary protein, serum triglyceride and total cholesterol increased significantly (all P<0.05), while the proportion of gross hematuria episodes and the ratio of serum albumin to globulin significantly decreased (all P<0.05). For the aspect of pathological manifestations, the proportions of cell proliferation in capillaries (E1), segmental sclerosis or adhesion (S1), renal tubular atrophy or interstitial fibrosis (T1/2), globular sclerosis, renal arteriole wall thickening and vitreous degeneration, Lee's grade Ⅳ and Ⅴ increased with the decrease of baseline renal function (all P<0.05). Kaplan-Meier analysis showed that the cumulative renal survival rate decreased with the decline of baseline renal function (Log-rank χ2=88.510, P<0.001). As a result of multivariate Cox regression analysis, nephrotic syndrome ( HR=2.399, 95% CI 1.054-5.459, P=0.037), hypertension ( HR=1.806, 95% CI 1.071-3.048, P=0.027), low baseline eGFR (taking group A as the reference, group B: HR=2.383, 95% CI 1.053-5.392, P=0.037; group C: HR=6.878, 95% CI 3.074-15.393, P<0.001), IgG deposition ( HR=2.224, 95% CI 1.384-3.574, P=0.001) and globular sclerosis ( HR=2.075, 95% CI 1.230-3.501, P=0.006) were the independent influencing factors for renal progression in primary IgAN patients. Conclusions:The level of baseline renal function in primary IgAN patients can be used to predict the extent of clinic-pathological damage. Nephrotic syndrome, hypertension, low baseline eGFR, IgG deposition and globular sclerosis are the independent influencing factors for renal progression in primary IgAN patients.

Objective To compare the incidence of cardiopulmonary complications between treatment of video assisted thoracoscopic operation and traditional thoracotomy operation in elderly patients with non-small cell lung cancer.Methods 122 cases of elderly patients with non small cell lung cancer were treated in our hospital from Mar 2010 to Jun 2014,and randomly divided into observation group (therapeutic lobectomy by video assisted thoracoscopic operation) and control group (therapeutic lobectomy by traditional thoracotomy operation) (n=61,each).The postoperative cardiopulmonary complications and relevant indexes of cardiopulmonary function were analyzed and compared between the two groups.Results Before operation,there was no significant difference in the relevant indexes of cardiopulmonary function between the two groups (P＞0.05).After operation,the forced expiratory volume in one second (FEV1) and maximal ventilatory volume (MMV) were higher and the heart rate was lower in observation group than in control group (t=3.682,2.243,5.677,all P＜0.05).The incidence of cardiopulmonary complication was lower in observation group than in control group (13.1％ vs.27.9％,x2 =4.075,P=0.044).Conclusions Video assisted thoracoscopic operation is better for the recovery of cardiopulmonary function,and can reduce cardiopulmonary complication in elderly patients with non-small cell lung cancer.

BACKGROUND:Human umbilical cord blood-derived mesenchymal stem cels can be induced through the co-culture to differentiate into other cels, but how to get more seed cels for tissue engineering is one of the most difficult problems. OBJECTIVE: To investigate the role of the different concentrations of thyroxine in chondrogenic differentiation of human umbilical cord blood-derived mesenchymal stem cels by co-culture with rabbit chondrocytes. METHODS:Human umbilical cord blood-derived mesenchymal stem cels were co-cultured with rabbit chondrocytes at 2:1, and stimulated by medium containing different concentrations of thyroxine (0, 0.01, 0.1 and 1, 10 μmol/L). Co-cultured cels with no thyroxine served as control group. After 14 days of co-culture, the cel RNA and protein were extracted, mRNA expressions of aggrecan and colagen type II were detected by real-time PCR, and protein expression of aggrecan and colagen type II were detected by western blot assay. RESULTS AND CONCLUSION:After intervention with 0.01, 0.1, 1, 10 μmol/L thyroxine, the mRNA and protein expressions of aggrecan and colagen type II were enhanced with the increase of thyroxine concentration, which were significantly different from those in the control group (P < 0.05). Experimental findings indicate that high levels of thyroxine can enhance the chondrogenic ability of human umbilical cord blood-derived mesenchymal stem cels co-cultured with rabbit chondrocytes.

Objective To investigate the infection status of human papillomavirus (HPV) of females in Chongqing city ,in or‐der to provide scientific references for prevention of HPV infection and control of early cervical lesions .Methods The detection re‐sults of cervical cytology screening for 19 860 females in the University‐Town Hospital of Chongqing Medical University ,the First Affiliated Hospital of Chongqing Medical University and a county‐level hospital in Chongqing from January 2012 to January 2015 were collected ,and the HPV typing data were analysed .Results Among the 19 860 females ,5 681 females were infected with HPV ,the infection rate was 28 .61% .Due to the presence of multiple infection ,the actual infection frequency of HPV subtypes in 5 681 females was 8 906 .The infection frequency of high‐risk subtypes ,including multiple infections ,was 6 166 (accounted for 69 .23% ) .According to the probability of infection ,the top eight rate of subtype infection was HPV 52 ,16 ,58 ,6 ,CP8304(81) ,33 , 53 and 18 .The positive rate of single HPV infection was 19 .38% ,the positive rate of multiple infection was 8 .74% ,in which doub‐le infection was the main infection model .The infection rate of females aged 20 years old or less(43 .89% ) was the highest ,while have the infection rate of females aged 36-50 years old was the lowest(27 .26% ) .Conclusion The HPV infection rate of females in Chongqing is high ,the most of females infected by high‐risk subtype HPV .It is necessary to lay emphasis on preventing HPV 52 , 16 ,58 ,33 ,18 infection for females in Chongqing city .

AIM: To construct prokaryotic expression vector of His-tagged human IP-10 for further study of its biological function in the inflammatory response. METHODS: The coding sequence of IP-10 lacking signal peptide was amplified from human lung cDNA library by polymerase chain reaction (PCR) and the fragment was cloned into pET-14b plasmid for the construction of His-tagged fusion protein expressing vector, pET-14b/IP-10. After being identified by enzyme digestion and sequencing, the recombinant vector was transformed into a strain of E. coli, BL21 (DE_3). The expression of His-tagged fusion protein was induced with IPTG and purified with Ni+-NTA affinity chromatography. Then the chemotactic activity of IP-10 was determined by transwell migration assay on THP-1 cells. RESULTS: The construction of pET-14b/IP-10 recombinant vector was proved by enzyme digestion and sequencing. The fusion protein IP-10, which was purified by a routine Ni+ affinity method, had an activity on the induction of cell migration of THP-1. CONCLUSION: We successfully construct IP-10 fusion protein expressing vector and get the fusion protein with high bioactivity, which provides essential materials for the future studies on IP-10.

Acute Kidney Injury ; epidemiology ; pathology ; Adolescent ; Adult ; Aged ; Biopsy, Needle ; methods ; China ; epidemiology ; Female ; Glomerulonephritis, Membranoproliferative ; epidemiology ; pathology ; Humans ; Kidney ; pathology ; Kidney Diseases ; epidemiology ; pathology ; Male ; Middle Aged ; Nephrotic Syndrome ; epidemiology ; pathology ; Ultrasonography, Interventional

Objective To determine the effect of NEL-like type 1 gene (NELL-1) transfection in vivo in the repair of traumatic femoral head necrosis.Methods Twenty-four SD rats were randomly divided into three groups (8 rats per group) according to the lottery method,ie,sham group (served as normal control),NELL-1 treatment group (injected NELL-1 gene by recombinant adenovirus vectors around the hip one week after osteonecrosis model induced surgically) and placebo group (given an equal volume of saline solution at the same time after the induction of osteonecrosis).Femurs were taken from the animals 5 weeks after surgery.Gross observation was performed for morphology changes,X-ray assessment for femoral head height and length ratio (H/L),Micro-CT measure for bone parameters of femoral head including total volume (TV),bone volume (BV),total mineralized content (TMC),trabecular thickness (Tb.Th) and trabecular space (Tb.SP),and histological study for osteocytes,osteoblasts and osteoclasts.Results Preserved femoral head shape was noted in NELL-1 treatment group compared to the obvious flattening of the femoral head in placebo group.No heterotopic osteogenesis was observed in any group.Femoral head H/L ratio for 0.753 2 ± 0.040 2 in NELL-1 treatment group was higher than 0.598 4 ± 0.037 0 in placebo group (P ＜ 0.05),but lower than 0.920 2 ± 0.037 0 in sham group (P＜0.05).TV,BV,TMC and BMD between NELL-1 treatment and sham groups did not differ significantly (P ＞ 0.05),but all were increased compared to placebo group (P ＜ 0.05).There was no significant differences in Tb.Th and Tb.SP among three groups (P ＞ 0.05).Most osteocytes were alive in NELL-1 treatment group.More active osteoblasts and osteoclasts were noted in NELL-1 treatment group than those in placebo group.Conclusion NELL-1 gene transfection can preserve femoral head shape and bone content,promote osteoblast activity and neovascularization and hence is an effective treatment for rat traumatic osteonecrosis.However,the activity of osteoclasts is stimulated simultaneously.