levels are elevated in patients with NAFL and correlated with HOMA-IR, suggesting RBP4 may participate in the development of IR and NAFL.

Objective To observe the clinical effect and safety of umbilical cord blood mesenchymal stem cell(UCB-MSC) transplantation in the treatment of diabetic foot. Methods UCB-MSC suspension (cell concentration (3 -7) × 107/L,0.3 -0.5 ml per point) was injected into multiple spots on affected lower limb with a 3 cm × 3 cm istance among each point. Demixing injection could be performed in regions with multilayer muscles. Clinical symptoms and related index were routinely observed from the first day to three months after operation. Results After three months of UCB-MSC transplantation, pain of patients was relieved, skin temperature increased, intermittent claudication ameliorated, ulcer healed, ankle-brachial index and transcutaneous partial pressure of oxygen increased. The lower extremity lesions showed an abundant collateral vessel formation after the treatment in 2 patients by angiography. Both patients had no severe complications and adverse reactions, none underwent amputation. Conclusions Umbilical cord blood mesenchymal stem cell transplantation is a safe and effective treatment of diabetic lower limb ischemic disease, which can exempt the patient from amputation and improve their quality of life.

BACKGROUND: Many animal and clinical studies have reported that the safe and effective usage of umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) transplantation for treatment of neurological genetic diseases.OBJECTIVE: To investigate the feasibility and effect of UCB-MSCs transplantation in the treatment of spinal muscular atrophy (SMA).METHODS: A child admitted at January 2010 had been confirmed as having SMA, and drug and rehabilitation therapies were invalid. Then, the child received UCB-MSCs transplantation via the first intravenous infusion and three times of subarachnoid injection, once a week, (4-6)×107 cells once and four times as a course. Neurological physical examination, biochemical test, muscle enzymes detection, FIM scoring and electromyography (EMG) examination were conducted. RESULTS AND CONCLUSION: Compared with prior to transplantation, the level of muscle enzymes decreased, FIM scores were increased from 68 to 93 points, EMG results showed that the motor units with re-contraction in each 10.0 ms were increased that the motor function was improved, the lower extremity muscle strength elevated, and the self-care ability was improved in the SMA child at 6 months after transplantation. During the 10-month follow-up, the child had no adverse effects. It is indicated that UCB-MSCs transplantation is effective to treat SMA, and the neurological function has a remarkable restoration.

Objective: To investigate the status of tuberculin skin test (TST) results and the influencing factors among student close contacts with pulmonary tuberculosis, so as to provide the evidence for developing prevention and control strategies for pulmonary tuberculosis among students. Methods: The students aged 15 years and above who had close contact with pulmonary tuberculosis cases in Yuecheng District, Shaoxing City, Zhejiang Province, from October 2016 to December 2023 were recruited and investigated using questionnaires and TST to collect demographic information, contact history, and TST results. The influencing factors for TST positivity among student close contacts with pulmonary tuberculosis were analyzed using a multivariable logistic regression model. Results: A total of 5 507 student close contacts were investigated, including 2 982 males and 2 525 females, with a male-to-female ratio of 1.18∶1. The mean age was (19.10±1.71) years. Among them, 397 (7.21%) were technical secondary school students, 766 (13.91%) were senior high school students, 2 556 (46.41%) were junior college students, and 1 788 (32.47%) were college students or above. A total of 958 students tested positive for TST, with a positivity rate of 17.40%. The rates of general positivity, moderate positivity, and strong positivity were 10.53%, 4.98% and 1.89%, respectively. Multivariable logistic regression analysis showed that senior high school students (OR=1.473, 95%CI: 1.009-2.152) and junior college students (OR=1.467, 95%CI: 1.074-2.005), as well as those with an exposure-to-screening interval of ≥46 days (46-<61 days, OR=2.043, 95%CI: 1.478-2.826; ≥61 days, OR=1.291, 95%CI: 1.018-1.637) had a higher risk of TST positivity. Female student close contacts had a lower risk of TST positivity (OR=0.753, 95%CI: 0.649-0.873). Conclusion The TST positivity rate was relatively high, and gender, school type, and exposure-to-screening interval were influencing factors for TST positivity among student close contacts with pulmonary tuberculosis.

OBJECTIVETo investigate the expression of high mobility group box 1 (HMGB1) in gingival tissues of chronic periodontitis.

METHODSHuman peripheral blood mononuclear cells(PBMC) were stimulated with 1 microg x mL(-1) lipopolysaccharide (LPS) for 24 h or 48 h. Expression and release of HMGB1 were checked by immunofluorescence and enzyme-linked immunosorbent assay (ELISA), respectively. PBMC were stimulated with 100 ng x mL(-1) HMGB1 or 50 ng x mL(-1) tumor necrosis factor-alpha (TNF-alpha), the expressions of TNF-alpha and HMGB1 in the supernatant were studied by ELISA. Gingival tissues and gingival crevicular fluids (GCF) were collected from patients and healthy people. Expression of HMGB1 in gingival tissues and GCF was studied using immunofluorescence and ELISA, respectively.

RESULTSHMGB1 was translocated from nucleus to cytosol in PBMC after LPS stimulation for 24 h. The content of HMGB1 in the supernatant from stimulated cells was significantly higher than that from unstimulated cells after 48 h (P < 0.01). HMGB1 was released by PBMC in response to TNF-alpha stimulation, it also stimulated PBMC to release TNF-alpha (P < 0.01). Translocation of HMGB1 from nucleus to cytosol was also found in infiltrated cells in gingival tissues from patients, and HMGB1 in GCF from patients was significantly higher than that from healthy people P < 0.01).

CONCLUSIONThe results suggest that HMGB1 may play an important role in the pathological progress of chronic periodontitis.

Enhanced green fluorescent protein( EGFP), myc epitope and polyhistidine metal-binding tag are often used as a marker for recombinant fusion protein in many gene expression vectors, each marker has its own function, EGFP emits green fluorescence for direct detection, myc epitope facilitates recombinant fusion protein detection using its antibodies, polyhistidine tag allows purification of recombinant fusion protein using resin.Hitherto, no a plasmid vector can integrate all of these functions. In this study we constructed a novel eukaryotic expressive plasmid, designated as pcDNA6/myc-his-EGFP B, which integrated the functions of EGFP, myc epitope and polyhistidine tag. Importantly, a linker octo - peptide in N terminal of EGFP was designed using LINKER program. A DNA fragment encoding a putative protein containing a signal peptide of human interleukin 2(IL-2) in N terminal was cloned into pcDNA6/myc-his-EGFP B in frame with the C-terminal peptide to construct pMHES. 2.2.15 Cells were transfected with pcDNA6/myc-his-EGFP B and pMHES, and Balb/c mice were intravenously injected with pcDNA6/myc-his-EGFP B by tails, results revealed that both of the plasmids worked in 2.2.15 Cells and livers of Balb/c mice. Assuming gene of the IL-2 was inserted into pcDNA6/myc-his -EGFP B in frame with EGFP, myc and 6 × His, three-dimensional structure for this putative expression product was simulated using Modeller8V2, results revealed that IL-2, EGFP, myc and 6 × his did not interfere each other and octo- peptide linker owned certain flexibility. The results suggest that pcDNA6/myc-his-EGFP B may be useful as a genetic tool for mammalian cells and a vector for gene therapy.

Objective To investigate the use and effect of dorsal dartos flap wrapping technique in complicated hypospadias fistulas repairs. Methods In the experimental group, the closed fistulas were wrapped with transferred dorsal dartos flap in the 32 cases of complicated hypospadias fistulas from March 2004 to January 2008. In the control group, the routine fistulas repairs were preformed in the 50 cases of complicated hypospadias fistulas from January 1998 to February 2004 . Results All cases had been followed up for 3 - 12 months, there were no fistula in the experimental group. There were 6 fistulas in the control group. There was significant difference between two groups (P<0.05). Conclusions The use of the technique can prevent urethrocutaneous fistulas after complicated hypospadias fistulas repairs, the fistulas rate are reduced for all cases combined. The technique would be the optimal repair for complicated hypospa-dias fistulas.

Objective To analyze videourodynamic characteristics of female bladder outlet obstruction (BOO). Methods Retrospective analysis of videourodynamic characteristics were performed in 42 women with BOO. On the basis of the videourodynamic findings, women with BOO were categorized into five groups: bladder neck obstruction, middle urethra (urethral sphincter) obstruction, distal urethra obstruction, external orifice of urethra obstruction, high-grade pelvic organ prolapse. Data of five groups were compared in Qmax, maximum cystometric capacity, Pdet at Qmax, residual urine, detrusor instability and hydronephrosis. Data of all patients were analyzed by SPSS13 statistical software, measurement data by t-test and enumeration data by x2-test. Results Five groups had not significant deviation in the maximum cystometric capacity. however patients with bladder neck obstruction had significant deviation with others in Pdet at Qmax (78. 0 ± 23. 6)cm H2O, residual urine (120. 6±115.2)ml, Qmax (7.0±2.3)ml/s (P<0.05). There was significant deviation between middle urethra (urethral sphincter) obstruction(76.9%)and others in detrusor instability(P<0. 05). In all patients with hydronephrosis, bladder neck obstruction(57. 1 %) had significant deviation with others (P<0. 05). Conclusions Videourodynamic tests could assess lower urinary tract function, and may guide clinic management. The bladder neck obstruction was most serious and harmful to upper urinary tract.

Objective: To observe the different effects between Mahuang (Herba Ephedra Sinicae) and Wuweizi (Fructus Schisandrae Chinensis) on the pathological changes of rats with bleomycin A(5)-induced pulmonary fibrosis. Methods: Ninety Wistar rats were randomly divided into sham operation group, model group, hydrocortisone group, Herba Ephedra Sinicae group, Fructus Schisandrae Chinensis group and Herba Ephedra Sinicae plus Fructus Schisandrae Chinensis group. There were 16 rats in each group except the sham operation group (10 rats). Pulmonary fibrosis was induced by a single intratracheal injection of bleomycin A5. Hematoxylin and eosin straining and immunohistochecical method were used after 7- and 28-day treatment to observe the pathology of lung injury, measure the inner diameter of pulmonary arterioles and the density of nuclear membrane. Results: Compared with the sham operation group at 7 and 28 d, alveolar inflammation level was significantly increased in the model group (P<0.01). Alveolar inflammation level was decreased obviously in the hydrocortisone group (P<0.05) after 7- and 28-day treatment as compared with the model group, and that in Herba Ephedra Sinicae plus Fructus Schisandrae Chinensis group was also decreased obviously (P<0.05) at 28 d. Compared with the sham operation group, nuclear density of the model group was increased, while its inner diameter was decreased (P<0.05). In the Fructus Schisandrae Chinensis group, Herba Ephedra Sinicae plus Fructus Schisandrae Chinensis group and hydrocortisone group, the nuclear density was decreased (P<0.05) as compared with the model group. Inner diameter in the Herba Ephedra Sinicae group, Herba Ephedra Sinicae plus Fructus Schisandrae Chinensis group and hydrocortisone group was higher than that in the model group (P<0.05). Microvessel density of the model group was obviously higher than that of the sham operation group (P<0.05). Compared with the model group, Herba Ephedra Sinicae plus Fructus Schisandrae Chinensis group and hydrocortisone group had lower microvessel density (P<0.05). Conclusion: Herba Ephedra Sinicae combined with Fructus Schisandrae Chinensis can restrain pulmonary artery injury. The nuclear density and microvessel density can be reduced by Fructus Schisandrae Chinensis, while Herba Ephedra Sinicae can increase the inner diameter.

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