Objective To estimate effects of AG490, a selective inhibitor of JAK2/STAT3 signaling pathway, on the biological behavior of human keloid?derived fibroblasts (HKFs), and to explore their possible mechanisms. Methods In vitro cultured human skin fibroblasts(HSFs)and HKFs were both divided into several groups to be treated with AG490 at different concentrations (12.5, 25, 50, 75, 100 μmol/L), with those receiving no treatment serving as the control group. Then, cell counting kit?8(CCK?8)assay was performed to evaluate cellular proliferative activity of HSFs and HKFs after 24?, 48?and 72?hour treatment, flow cytometry to estimate cell cycle distribution and apoptosis rate in HKFs after 24?hour treatment, reverse transcription(RT)?PCR to measure STAT3 and cyclin D1 mRNA expressions in treated HKFs as well as STAT3 mRNA expression in untreated HSFs and HKFs after 24?hour culture, and Western blot analysis to measure the protein expressions of STAT3 and p?STAT3 in HSFs and HKFs after 24?hour treatment. Results CCK?8 assay showed that the proliferation inhibition rates of both HSFs and HKFs gradually increased along with the increase in AG490 concentrations and treatment duration, and the inhibitory effects increased in both dose?and time?dependent manners(all P<0.05). Besides, when cells were treated with the same concentrations of AG490 for same durations, the proliferation of HKFs were inhibited to a greater extent than that of HSFs(all P<0.05). As flow cytometry revealed, along with the increase of AG490 concentrations, the proportion of HKFs in G1 phase and the apoptosis rate in HKFs both increased gradually(all P < 0.01), while the proportion of HKFs in G2 phase gradually decreased(all P < 0.01), and the proportion of HKFs in S phase remained insignificantly changed. RT?PCR showed that the mRNA expression of STAT3 was significantly higher in untreated HKFs than in untreated HSFs after 24?hour normal culture(P < 0.05). After 24?hour treatment with AG490, the mRNA expressions of STAT3 and cyclin D1 in HKFs gradually decreased with the increase of AG490 concentrations. Correlation analysis revealed that the mRNA expression of cyclin D1 was positively correlated with that of STAT3 in AG490?treated HKFs (r = 0.855, P < 0.01). Western blot analysis showed that the protein expressions of both STAT3 and p ? STAT3 gradually decreased in HKFs and HSFs along with the increase of AG490 concentrations(all P < 0.05), and were significantly lower in HKFs than in HSFs (both P < 0.05). Conclusion AG490 can effectively inhibit HKF proliferation by selectively blocking the JAK2/STAT3 signaling pathway.

BACKGROUND: Apoptosis secondary to ischemia and hypoxia is the main cause of spinal cord dysfunction. Because of the decrease in atmospheric pressure, patients living on the Qinghai-Tibet Plateau are in a hypoxic environment, which is very unfavorable for the recovery of spinal cord injury. Hyperbaric oxygen therapy can improve the postoperative function of patients with incomplete spinal cord injury, and its effect is better on the plateau than at normal altitudes.OBJECTIVE: To observe the effect of hyperbaric oxygen therapy on traumatic spinal cord injury in patients living on the Qinghai–Tibet Plateau.METHODS: This prospective, open-label, randomized controlled clinical trial was performed at the Department of Spine Surgery, Affiliated Hospital of Qinghai University, China. In total, 164 patients with incomplete traumatic spinal cord injury were equally and randomly assigned to a control group and a hyperbaric oxygen therapy group. Patients in the control group were treated with pedicle screw fixation and decompressive laminectomy. In addition to the surgical treatment performed in the control group, patients in the hyperbaric oxygen group underwent hyperbaric oxygen therapy at 0.2 MPa once a day for four treatment courses. Ten treatment sessions constituted one course, and each course was separated by a 5- to 7-day rest interval. The primary outcome was the modified Barthel index to assess activities of daily living. The secondary outcomes were the American Spinal Injury Association (ASIA) impairment scale grade, sensory score, and motor score. The study protocol was approved by the Ethics Committee of the Affiliated Hospital of Qinghai University, China (Approval number: QHC011K). Written informed consent was provided by a relative or legal representative of each patient after they had indicated that they fully understood the treatment plan. RESULTS AND CONCLUSION: The partial results demonstrated that after four treatment courses (55-61 days), the modified Barthel index and ASIA tactile, pain, and motor scores were higher in the hyperbaric oxygen group than in the control group. The ASIA grades were significantly different between the hyperbaric oxygen group and control group. The proportion of patients with ASIA grades D and E was higher in the hyperbaric oxygen group than in the control group. In this trial, we aim to determine the efficacy of hyperbaric oxygen therapy on the treatment of incomplete traumatic spinal cord injury in patients living on the plateau and to provide clinical evidence for treating incomplete traumatic spinal cord injury in these patients.

Objective To understand the clinical features of acute renal failure(ARF)as the initial presentation of systemic lupus erythematosus(SLE).Methods Eight cases of ARF in SLE from Jan 1995 to Apt 2006 were investigated,descriptive analysis and literature review were performed.Results①The symp- tom of ARF in SLE was mainly oliguria,with severe accompany symptoms and complications.②The level of leucocyte and hemoglobin was low in laboratory tests,also the complement level decreased significantly.The most frequent renal pathology was typeⅣ,Ⅳ+ⅤLN.③Large dose steroid and CTX were the mainstay of treatment.In addition,SCUF,CVVHDF and hemodialysis could be used for lethal conditions.Conclusion ARF can be the first manifestation of SLE and it usually represents more severe disease and more complica- tions.Large dose steroid and CTX can improve prognois.In cases refractory to steroid and if the effect is obso- lete,CTX treatment SCUF,CVVHDF and hemodialysis can be use.

OBJECTIVETo explore the clinical application value of Provider-Initiated HIV Testing and Counseling (PITC) by analyzing the positive rate of HIV tests for people in need of PITC and that of routine HIV tests.

METHODSWe retrospectively analyzed the demographic and epidemiologic data about the patients seeking PITC services or undergoing routine HIV tests in Nanjing Drum Tower Hospital between January and December 2013.

RESULTSThe positive rate of initial HIV screening was 1.98% in the PITC group and 0.24% in the routine test group, while that of confirmed HIV was 0. 40% in the former and 0.07% in the latter, both with statistically significant differences between the two groups (P < 0.01). The positive rate of HIV was markedly higher in males than in females, particularly in the PITC group.

CONCLUSIONPITC has a high clinical value in HIV detection for targeted subjects and therefore deserves general application in dermatology.

Counseling ; Dermatology ; Female ; HIV Seropositivity ; diagnosis ; epidemiology ; Humans ; Male ; Mass Screening ; methods ; Retrospective Studies ; Sex Factors

Objective To investigate the inhibitory effects of rosiglitazone on the synthesis of reactive oxygen species (ROS) and the expression of monocyte chemoattractant protein 1 (MCP-1) induced by high glucose in rat mesangial cells. Methods The mesangial cells were divided into six groups: control group ( C, 5.6 mmol/L glucose), mannitol group (M, 24.2 mmol/L mannitol+group C), high glucose group( H, 30 mmol/L glucose), R1 group(R1, group H+10 μmol/ L rosiglitazone), R2 group (R2, group H+20 μmol/L rosiglitazone), N-acetylcysteine (NAC) group (N, group H+5 mmol/L NAC, NAC was added 1 h before the stimulation of high glucose). The level of ROS was measured by confocal laser scanning microscopy. The mRNA and the protein expression of MCP-1 were semi-quantitatively determined with reverse transcription-polymerase chain reaction and ELISA respectively. Results No significant differences of ROS and MCP-1 were found between control group and mannitol group. The intracellular ROS induced by high DOI:10.3760/cma.j.issn. 1001-7097.2009.01.011glucose increased by 4.1-fold compared to control group (P<0.01), which was prevented by rosiglitazone (20 μmol/L) and NAC respectively. The MCP-1 mRNA expression in group R2 and group N was significantly lower than that in group H (P<0.01). The MCP-1 protein level in group H [(940.9±20.3) ng/L] was higher than that in group C [(403.0±8.1) ng/L] (P<0.01), and the expression of MCP-1 protein in group R2 [(562.5±15.3) ng/L] and group N [(539.8±8.3) ng/L] was lower than that in group H (P<0.01). Conclusion Rosiglitazone may suppress high glucose-induced MCP-1 expression by reducing the level of ROS, which may be one of the mechanisms that rosiglitazone plays a direct role in the protection of kidney.

Objective To study the effect of paeonol on the proliferation and apoptosis of A375 human melanoma cells and its mechanism.Methods Cell counting kit-8 (CCK8) was used to evaluate the proliferative activity of A375 cells treated with paeonol of 0.5,1,2,4,8 mmol/L for 24,48 and 72 hours respectively.Subsequently,A375 cells were treated with paeonol of 1.25,2.5 and 5 mmol/L for 24 hours followed by double staining with annexin V and propidium iodide for the detection of cell apoptosis,fluorometric assay for the estimation of caspase 3,caspase 8 and caspase 9 activity,and Western blot for the determination of the levels of p53,nuclear factor-κB proteins and some of their target proteins.The A375 cells receiving no treatment served as the blank control group.Statistical analysis was carried out by t test.Results Within the investigated concentration and time ranges,paeonol significantly inhibited the proliferative activity of A375 cells in a concentration-and timedependent manner.Compared with the blank control group,a significant increase was observed in the early apoptosis rate in A375 cells treated with paeonol of 1.25,2.5 and 5 mmol/L for 24 hours (13.74％-± 1.73％,25.95％ ± 0.57％ and 46.44％ ± 0.81％ vs.3.11％ ± 0.53％,P ＜ 0.05 or 0.01),as well as in the activity of caspase 3,8 and 9 in A375 cells treated with paeonol of 2.5 and 5 mmol/L for 24 hours (P ＜ 0.05 or 0.01).After 24-hour treatment,the protein levels of p53 and Bax were elevated,but those of nuclear factor-κB,Bcl-2 and Bcl-XL were decreased in A375 cells with the increase of paeonol concentration.Conclusions Paeonol can inhibit the proliferation but induce the apoptosis of A375 cells,and the apoptosis-inducing effect may be realized through intrinsic and extrinsic pathways by modulating nuclear factor-κB and p53 genes.

OBJECTIVETo observe anti-cancer effects of Jianpi Jiedu Recipe (JJR) on liver cancer (LC) rats with Pi deficiency syndrome (PDS) and its relation with the third complementary-determining region gene spectratyping of TCRVβ-chain (TCRVβCDR3).

METHODSRats were divided into 8 groups according to random digit table, i.e., the blank control group (normal), the PDS group, the LC model group, the LC-PDS group, high, middle, and low dose JJR groups (75.00, 37.50, 18.75 g/kg, respectively by gastrogavage, once per day), the thymus pentapeptide group (5 mg/kg, intramuscular injection, twice per week), 8 in each group. Rats in the normal group were administered with physiological saline by gastrogavage once per day. PDS rat model was prepared by bitter-cold purgation. LC model was prepared by orthotopic transplantation method. Twenty gene subfamilies of TCRβCDR3 in the thymus, liver, and LC tissues were detected by Gene Scan.

RESULTSHigh and middle dose JJR could postpone the growth of LC volume (P < 0.05), with equivalent liver index and thymus index to those of the normal group (P > 0.05). In thymus and liver tissue of the normal group, the number of clones (20 and 19), gene fragment number (220 and 113), Quasi-Gaussian distribution ratio of TCRVβCDR3 gene repertoire (100.0% and 42.1%), and fragment fluorescence peak area (6,539 ± 2,325 and 1,238 ± 439) were at the highest level among the 8 groups. TCRVβCDR3 expressions in thymus and liver tissue of high and middle dose JJR groups were approximate to those of the normal group. They were in the middle of the thymus pentapeptide group, the PDS group, the LC model group, and poorest in the LC-PDS group. TCRVβCDR3 in liver tissue expressed the best in the thymus pentapeptide group.

CONCLUSIONJJR might inhibit the growth of LC cells, and its mechanism might be related to enhancing TCRVβCDR3 spectratype expression.

Animals ; Complementarity Determining Regions ; genetics ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation, Neoplastic ; Genes, T-Cell Receptor beta ; Liver Neoplasms ; drug therapy ; genetics ; Random Allocation ; Rats

Objective To evaluate the effect of electric vagal stimulation on postoperative cognitive dysfunction in aged rats.Methods Thirty healthy Sprague-Dawley rats of both sexes,aged 18-20 months,weighing 390-550 g,were randomly divided into 3 groups (n=10 each) using a random number table:control group (group C),surgery group (group S),and electric vagal stimulation group (group V).Incision of the right carotid sheath was performed,and the vagal nerve was exposed after anesthesia in S and V groups.The right vagus nerve was stimulated for 30 min with continuous electric rectangular pulses (1 ms,10 Hz,1-2 V) after surgery in group V.The Morris water maze place navigation test was performed at 4,3,2 and 1 days before surgery and 2 days after surgery.The cognitive function was assessed using spatial probe and open field tests conducted at 2 days after surgery.The escape latency,the number of crossing the platform,the number of crossing the grid,the number of standing on the back legs,and the time animals spent in the central square were recorded.After the end of behavioral testing,venous blood samples were collected from the jugular vein for determination of concentrations of serum tumor necrosis factor-alpha (TNF-α),interleukin (IL)-1β and IL-6 using enzyme-linked immunosorbent assay.Results Compared with group C,the number of crossing the platform,the number of crossing the grid,and the number of standing on the back legs were significantly decreased,the escape latency and the time animals spent in the central square were significantly prolonged,and the concentrations of serum TNF-α,IL-1β and IL-6 were significantly increased at 2 days after surgery in group S (P＜0.05).Compared with group S,the number of crossing the platform,the number of crossing the grid,and the number of standing on the back legs were significantly increased,the escape latency and the time animals spent in the central square were significantly shortened,and the concentrations of serum TNF-α,IL-1β and IL-6 were significantly decreased at 2 days after surgery in group V (P＜0.05).Conclusion Electric vagal stimulation can improve the postoperative cognitive dysfunction in aged rats.

Objective To evaluate the value of reconstruction of three-dimentional CT in development dislocation of hip(DDH)in children.Methods Twelve cases of DDH concluded 4 bilateral and 8 unilateral cases.To sum up,16 sick hips were operated and 8 normal hips were also obtained by three-dimensional CT(Hip speed Fi/x,GE Co).Results 3D reconstruction were used to show femoral head,(acetabulum) and relationship of acetabulum and femoral head respectively.The difference between FNA measurement of sick hips and those of normal hips were significant(P