This study was purposed to investigate the angiogenesis-promoting activities of human mesenchymal stem cells (hMSCs) modified by hepatocyte growth factor (HGF) and the underlying mechanisms. The hMSCs were transfected by recombinant adenoviral vector carrying human HGF gene and seeded onto the chicken chorioallantoic membrane. Three days later, the number of blood vessels was counted and their angiogenic response was compared with those of hMSCs of same generation, recombinant basic fibroblast growth factor (bFGF) and alpha-MEM as control. The expression levels of bFGF, VEGF, angiopoietin-1 and angiopoietin-2 were evaluated by RT-PCR assay. The results showed that gene-modified hMSCs exhibited greatest activity to promote angiogenesis while the angiogenic response was nearly same between groups treated by hMSCs and bFGF, all of which were significantly higher than that observed in control (p < 0.01). RT-PCR analysis revealed that hMSCs constitutively expressed multiple angiogenesis-associated growth factors and their levels seemed up-regulated by HGF gene transfer. It is concluded that HGF gene-modified hMSCs show a potent angiogenesis-promoting function and may be useful in the treatment of ischemic disorders.
Animals ; Cells, Cultured ; Chick Embryo ; Chickens ; Hepatocyte Growth Factor ; genetics ; Humans ; Mesenchymal Stromal Cells ; Neovascularization, Physiologic ; genetics ; Transfection

Female ; Hepatitis C ; complications ; Humans ; Interferon-alpha ; therapeutic use ; Liver Cirrhosis ; complications ; drug therapy ; etiology ; Middle Aged ; Recombinant Proteins ; therapeutic use ; Vitiligo ; complications ; drug therapy

OBJECTIVE: To investigate the effect of oxidative stress on the accumulation of heat shock protein 70 (HSP70) within C2C12 myogenic cells. METHODS: Heat shock response (42 degrees C for 1 h and recovery for 12 h at 37 degrees C) was used to induce the expression of heat shock protein 70. We constructed a recombinant plasmid of HSP70 with enhanced green fluorescent protein (EGFP). After being transfected transiently into C2C12 cells, immunoblotting was used to detect the expression of HSP70 induced by heat shock response and transfection. Immunocytochemistry, fluorescent microscopy and immunoblotting were used to detect the translocation of HSP70. RESULTS: Immunoblotting showed that the overexpression of HSP70 was induced by heat shock response and transient transfenction. HSP70 localized within the cytoplasm of the normal cells, but HSP70 translocated from the cytoplasm to the nucleus and the nucleolus at 1 h after the treatment of oxidative stress (0.5 mmol/L H2O2) by using immunocytochemistry, fluorescent microscopy and immunoblotting for cellular partial proteins. CONCLUSION Oxidative stress may induce the accumulation of heat shock protein 70 within the nucleolus.
Cell Nucleolus ; metabolism ; Cells, Cultured ; HSP70 Heat-Shock Proteins ; metabolism ; Humans ; Myoblasts ; cytology ; metabolism ; Myocytes, Cardiac ; cytology ; metabolism ; Oxidative Stress ; physiology

OBJECTIVE: To investigate the expression of c-fos in rats' skin during wound healing. METHODS: Immunohistochemistry was conducted on paraffin section from incised wounding model of rat skin. RESULTS: Fos protein improved from the time of 10 min after wounding in the wound edge, then it reached peak at 3 h. 24 h after injury, the quantity of Fos expression had no difference with that of normal skin. CONCLUSION Fos is sensitive after wound, but should be used with other criteria in wounding interval estimation as it's unstediness.
Animals ; Genes, Immediate-Early ; Immunohistochemistry ; Male ; Proto-Oncogene Proteins c-fos/biosynthesis* ; Rats ; Rats, Sprague-Dawley ; Skin/metabolism* ; Time Factors ; Wounds and Injuries/metabolism*

Adolescent ; Adult ; Drug Combinations ; Humans ; Injections ; Male ; Pyridoxine ; administration & dosage ; pharmacokinetics ; Pyrrolidonecarboxylic Acid ; administration & dosage ; pharmacokinetics

OBJECTIVE: To study the clinical features of severe type 7 adenovirus pneumonia in children. METHODS: A retrospective analysis was performed for the clinical data of children who were diagnosed with severe type 7 adenovirus pneumonia from February to June, 2019. RESULTS: Among the 45 children, the male/female ratio was 3:2 and the median age was 14 months. All children had repeated fever, cough, and pulmonary moist rales, and the mean duration of fever was 14±4 days. The median time from fever to dyspnea was 8 days, and the time from fever to mechanical ventilation was 11.6±2.5 d. There was no significant increase in white blood cell count, with neutrophils as the main type. There were slight reductions in hemoglobin and albumin, while platelet and fibrinogen remained normal. There were increases in aspartate aminotransferase, lactate dehydrogenase, procalcitonin, and C-reaction protein. The detection rate of mixed pathogens was 84%. Effusion in both lungs was the major change on chest imaging (64%). Bronchoscopic manifestations were endobronchitis, tracheomalacia, and plastic bronchitis. The incidence rate of respiratory complications was 100%, and extrapulmonary complications mainly involved the circulatory system (47%), digestive system (36%), and nervous system (31%). Among the 45 children, 16 were administered with 400 mg/kg intravenous immunoglobulin (IVIG) daily for 5 days, with a mean duration of fever of 16±5 days, and 29 were administered with 1 g/kg IVIG daily for 2 days, with a mean duration of fever of 13±4 days; there was a significant difference in the mean duration of fever between the two groups (P=0.046). The overall mortality rate was 11%. CONCLUSIONS Severe type 7 adenovirus pneumonia in children has severe conditions, with a high incidence rate of complications and a high mortality rate, so it should be diagnosed and treated as early as possible.
Adenoviridae ; Bronchitis ; Female ; Fever ; Humans ; Infant ; Male ; Pneumonia, Viral ; Retrospective Studies

BACKGROUNDThere are different materials used for anterior cruciate ligament (ACL) reconstruction. It has been reported that both autologous grafts and allografts used in ACL reconstruction can cause bone tunnel enlargement. This study aimed to observe the characteristics of bone tunnel changes and possible causative factors following ACL reconstruction using Ligament Advanced Reinforcement System (LARS) artificial ligament.

METHODSForty-three patients underwent ACL reconstruction using LARS artificial ligament and were followed up for 3 years. X-ray and CT examinations were performed at 1, 3, 6, 12, 24, and 36 months after surgery, to measure the width of tibial and femoral tunnels. Knee function was evaluated according to the Lysholm scoring system. The anterior and posterior stability of the knee was measured using the KT-1000 arthrometer.

RESULTSAccording to the Peyrache grading method, grade 1 femoral bone tunnel enlargement was observed in three cases six months after surgery. No grade 2 or grade 3 bone tunnel enlargement was found. The bone tunnel enlargement in the three cases was close to the articular surface with an average tunnel enlargement of (2.5 ± 0.3) mm. Forty cases were evaluated as grade 0. The average tibial and femoral tunnel enlargements at the last follow-up were (0.8 ± 0.3) and (1.1 ± 0.3) mm, respectively. There was no statistically significant difference in bone tunnel width changes at different time points (P > 0.05). X-ray and CT measurements were consistent.

CONCLUSIONSThere was no marked bone tunnel enlargement immediately following ACL reconstruction using LARS artificial ligament. Such enlargement may, however, result from varying grafting factors involving the LARS artificial ligament or from different fixation methods.

Adult ; Anterior Cruciate Ligament ; diagnostic imaging ; surgery ; Anterior Cruciate Ligament Reconstruction ; methods ; Female ; Humans ; Male ; Middle Aged ; Radiography ; Reconstructive Surgical Procedures ; methods ; Transplantation, Autologous ; Transplantation, Homologous ; Young Adult

BACKGROUNDA new fluroquinolone antibacterial agent, antofloxacin hydrochloride, developed in China, is an 8-NH(2) derivant of levofloxacin. The purpose of the study was to evaluate the pharmacokinetic characteristics of single and multiple oral doses of antofloxacin hydrochloride in Chinese healthy male volunteers.

METHODSAn open-label, non-randomized, single and multiple dose clinical trial was conducted. In single dose study, 12 subjects took 200 mg antofloxacin hydrochloride. In multiple dose study, 12 subjects took antofloxacin hydrochloride 400 mg once on day 1 and 200 mg once daily from day 2 to day 7. HPLC was used to assay the serum and urinary concentrations of antofloxacin.

RESULTSIn single dose study, the maximum concentration of drug in serum (C(max)), the time to reach C(max) (T(max)), and the area under the serum concentration-time curve (AUC (0-∞)) of antofloxacin were (1.89 ± 0.65) mg/L, (1.29 ± 0.26) hours, and (25.24 ± 7.26) mg×h(-1)×L(-1), respectively. Accumulating elimination rate of antoflocaxin from urine within 120 hours was 39.1%. In multiple dose study, blood concentration of antofloxiacin achieved stable state on day 2 after dosing. The minimum concentration drug in serum (C(min)), AUCss, mean concentration of drug in serum (C(av)), and degree of fluctuation (DF) were (0.73 ± 0.18) mg/L, (47.59 ± 7.85) mg×h(-1)×L(-1), (1.98 ± 0.33) mg/L, and 1.74 ± 0.60, respectively. On day 7 after dosing, T(max), C(max), and AUC (0-∞) was (1.14 ± 0.50) hours, (2.52 ± 0.38) mg/L, and (48.77 ± 8.44) mg×h(-1)×L(-1), respectively. Accumulating elimination rate of antofloxaxin from urine within 120 hours after the last dosing was 60.06%.

CONCLUSIONSThe regimen of 400 mg loading dose given on the first treatment day and then 200 mg dose once daily results in satisfactory serum drug concentration.

Administration, Oral ; Adolescent ; Adult ; Anti-Bacterial Agents ; administration & dosage ; blood ; pharmacokinetics ; urine ; Chromatography, High Pressure Liquid ; Humans ; Levofloxacin ; Male ; Ofloxacin ; administration & dosage ; analogs & derivatives ; blood ; pharmacokinetics ; urine ; Young Adult

·AIM:To explore the effectiveness of a new non-inverted pedicle internal limiting membrane ( ILM ) flap transposition technique in the treatment of large macular holes (MH). ·METHODS: This was a prospective pilot study which included 9 patients with 10 eyes in Jiangsu Province People's Hospital from December 2016 to February 2017. All patients was diagnosed with large MH (size >400μ m) by the spectra- domain optical coherence tomography (SD-OCT) and underwent the non-inverted pedicle ILM flap transposition surgery. Best-corrected visual acuity (BCVA), SD-OCT images, and MP-1 microperimetry tests were performed pre-operation, 1d, 1wk, 1, 3, and 6mo post-operation. ·RESULTS:The macular hole closure rate after 6mo was 100%. The averaged BCVA improved from 1. 19 ± 0. 54 (LogMAR) pre-operation to 0.70 ± 0.50 (LogMAR) post-operation (P=0.005). The mean retinal sensitivity within 8° and 2° improved from 3.14±4.52dB to 8.91±5.53dB(P=0.008), and 1.46 ± 2.94dB to 6.33 ± 4.90dB (P=0.008) respectively. Preoperative unstable fixation in seven eyes resolved at the last postoperative follow-up.·CONCLUSION: Our non-inverted pedicle internal ILM flap transposition technique shows effectiveness in the treatment of large macular holes with high MH closure rate and improving visual function.

Objective To construct the pEGFP-C1-CXCL1 eukaryotic expression vector and to investigate the effect of CXCL1 on the proliferation of HepG2 cells under endoplasmic reticulum stress ( ERS).Methods Fragments of CXCL1 were obtained from the cDNA library of HepG2 cells before CXCL1 was cloned into a pEGFP-C1 vector for a recombinant plasmid pEGFP-C1-CXCL1 which was screened and identified by PCR and sequence alignment .Then,the recombinant plas-mid of pEGFP-C1-CXCL1 was transfected into human 293 T cell line and the expression of CXCL 1 was detected by fluores-cence microscopy and Western blotting.pEGFP-C1-CXCL1was furhter transfected into HepG2 cells, and CCK8 was used to detect the inhibitory effect of CXCL1 on tumor proliferation induced by TM in hepatocellular carcinoma .Results pEGFP-C1-CXCL1 was vertified by sequencing analysis .Fluorescence microscopy showed that pEGFP-C1-CXCL1 was transfected into 293T.CXCL1 expression was detected by Western blotting .CCK8 showed that TM inhibited tumor proliferation , while overexpression of CXCL1 decreased the inhabitory rate on cell proliferation of HepG 2 cells under ER stress compared to pEGFP-C1 group and the control group .Conclusion A recombinant pEGFP-C1-CXCL1 plasmid is successfully constructed that can be expressed stably in human 293T cells.Overexpression of CXCL1 can effectively reduce the inhabitory rate of HCC cells induced by the ER stress.