The internal audit is important for the management system of laboratory .The article stated how to effectively implement internal audit in drug testing laboratories in terms of the planning , checklist, pattern, problem observation , CAPA of non-compliance and application of results in internal audit .

[Objective] To estimate the effects of ginsenoside Rb1 on melanogenesis in human melanocytes and underlying mechanisms.[Methods] Epidermal melanocytes were obtained from circumcision specimens of children,and subjected to primary culture.After 2 to 5 passages,the melanocytes were treated with different concentrations of ginsenoside Rb1,dimethyl sulfoxide (DMSO,vehicle control),forskolin at 10 μmol/L(positive control) or remained untreated (blank control).After additional culture for 72 hours,methyl thiazolyl tetrazolium (MTT) assay and NaOH lysis method were used to evaluate cell viability and melanin content in melanocytes respectively,spectrophotometer to determine dopa oxidase activity of tyrosinase,Western blot to quantify the protein level of tyrosinase,microphthalmia-associated transcription factor (MITF),phosphorylated and total cAMP response element binding protein (p-CREB and t-CREB) in melanocytes.[Results] After treatment with ginsenoside Rbl of 25,50 and 100 μmol/L for 72 hours,the melanocytes experienced no significant changes in viability (P ＞ 0.05 ),but a significant dose-dependent increase in melanin content (112.4％± 5.7％,155.7％ + 6.3％,217.2％ ± 11.7％ vs.100％,P＜ 0.05 or 0.01) and tyrosinase activity(117.9％ ± 5.7％,158.2％ ± 9.6％,182.6％ ± 10.0％ vs.100％,P＜ 0.05 or 0.01 ) compared with the vehicle control melanocytes.The protein expressions of tyrosinase,MITF and p-CREB were statistically higher in melanocytes treated with ginsenoside Rb1 of 100 μmol/L for 72 hours than in the vehicle control melanocytes (225.4％ ± 12.8％ vs.100％ ± 7.9％,313.5％ ± 16.7％ vs.100％ ± 9.8％,322.5％ ± 21.1％ vs.100％ ± 9.1％,all P＜ 0.01).The increase in MITF protein expression was inapparent in melanocytes at 8 hours after the treatment with ginsenoside Rb1 of 100 μmol/L,but statistically significant at 24 hours compared with the melanocytes at baseline (P＜ 0.01).The pretreatment with H-89 (a 8elective inhibitor of PKA) at 10 μmol/L,significantly suppressed the ginsenoside Rb1 (100 μmol/L for 72 hours) -induced phosphorylation of CREB,increase in MITF,tyrosinase expression,as well as tyrosinase activity and melanin content in melanocytes (all P ＜ 0.01 ).[Conclusion]s Ginsenoside Rb1could enhance the melanogenesis and tyrosinase activity in normal human melanocytes.The PKA/CREB/MITF/ tyrosinase signaling pathway may contribute to the pro-melanogenic effect of ginsenoside Rb1.

Objective To investigate the effects of human amniotic membrane on the survival of the crossboundary perforator flap in rats through distal subcutaneous implantation.Methods From February,2016 to December,2016,38 SD rats weighing 270-300 g were randomly divided into experimental group and control group (n=19).A three-territory perforator flap (3 cm× 10 cm) with the iliac artery as the pedicel elevated on the right dorsum of the rats.Experiment group,a section of amnion was sutured to the distal subcutaneous of the flap and then the flap was sutured back to its donor site.Control group,after the flap elevated,directly sutured back to its donor site.The blood flow of the pedicel were detected by laser Doppler flowmetry at the day before surgery,1 day after surgery,3 and 7 days after surgery,with 3 rats in every group.When the rats were anesthetized,biopsies were taken from the choke area Ⅱ at the day before surgery,3 and 7 days after surgery,with 9 rats in every group.When the rats were anesthetized,HE was used to compare the diameter size of the artery and vein at the same site.At day 7,measure the survival area of the flap,and 3 of them observe the vessel of the flap by lead oxide-gelatine technique.Results The survival rate of the experimental group and the control group after 7 days were (89.09±4.23)％ and (74.56-±5.59)％ respectively,the experimental group was significantly higher than that of the control group (t=5.48,P=0.00).X-ray showed that 7 days after operation,the pedicel of iliac artery in the experimental group was bigger than that in the control group.The blood flow detection showed that the blood flow of the experimental group was higher than that of the control group (t=2.39,3.06;P=0.03,0.00,respectively) at day 1 and day 3 after surgery.Hematoxylin eosin staining showed that arteriovenous tube diameter at the Choke Ⅱ were gradually increased on the day3 and day7 after surgery in both group,but the experimental group diameter expanded multiple was higher than that of control group (t=3.52,3.50;P=0.02,0.02,respectively).Conclusion Human Amnion subcutaneous embedding may improve the blood flow of the vascular pedicle,expand the microvascular at Choke area Ⅱ,improve microcirculation,as a result to promote the survival of cross perforator flap,while the mechanism is needed to understanding.

Objective To investigate the effects of DMOG on the microcirculation of the choke-area and the survival of the cross-boundary flap in rats via tail vein injection.Methods Rats with ischemic three-territory perforator flaps on the dorsum were treated with DMOG at a dosage of 40 mg/kg body weight via tail vein injection at 1 day before surgery(day-1),the time of surgery(day 0),1 day after surgery(day 1),2 days after surgery(day 2) and 3 days after surgery(day 3).Control group received sterile saline at the same time points and same dosage via tail vein injection.① Draw materials from the choke-area at day 1,day 3 and day 7,HE stain was used to compare the diameter size of the artery and vein at the same site.② Western blotting to check the expression of PCNA and HIF-1α,ELISA to detect the content of PCNA,HIF-1α,SDF-1α and VEGF at day 7.③At day 7,measure the survival area of the flap and observe the vessel of the flap by lead oxide-gelatine technique.Results ① There was a greater survival rate of (96.3 ± 5.1)％ in the treatment group than in the control group with (73.9 ± 5.8)％ at day 7 (P ＜ 0.05).② The diameter size of the arterioles and venules were dilated in both groups until postoperative days 7.But the treatment group was more expanded than the control group at day 3(2.20 ± 0.26 vs.1.50 ± 0.20,P ＜ 0.05) and day 7(3.67 ± 0.35 vs.2.03 ± 0.15,P ＜ 0.05).③ The skin expression of PCNA and HIF-1α in the treatment group were greater than the control group(P ＜ 0.05) at day 7.④ The content of skin PCNA in the treatment group and control group were(8.95 ± 0.71) ng/mg and (4.15 ± 0.72) ng/mg,HIF-1α were(5.04 ± 0.50)ng/mg and (2.98 ± 0.29) ng/mg,SDF-1α were (2.91 ± 0.61) ng/mg and (1.39 ± 0.62) ng/mg,and VEGF were(2.17 ± 0.41) ng/mg and (0.95 ± 0.44) ng/mg,respectively.The treatment group was greater than the control group (P ＜ 0.05).Conclusion DMOG can improve the microcirculation of the choke area,and then increase the survival of the perforator skin flaps in rats via tail vein injection.

Objective To explore the value of post processing technique of dual source CT in diagnosing peripheral PE.Methods One hundred and fifty patients with suspected PE were underwent dual source CT scan,and 20 of the all patients met the criterion. The raw CT data were reconstructed by the pulmonary embolism detection (PED)software,double energy perfusion imaging (DEPI)and CT pulmonary artery angiography (CTPA).The PED,DEPI and CTPA map were analyzed by 2 senior doctors in cardiovascular diagnosis profession.According to the diagnosis standard of PE,the location,number of the emboli in segmental and sub-segmental pulmonary arteries were respectively recorded on the PED and DEPI map.We calculated the detection rate,evaluated the significant difference and evaluated the diagnosis consistency between the DEPI and CTPA map.Results Emboli were found in 30 segmental and 40 sub-segmental arteries on the CTPA map with the detection rate of 7.50% and 5.00%,respectively.Emboli were found in 48 segmental and 62 sub-segmental arteries on PED map with the detection rate of 12.00% and 7.75%,respectively.There was significant difference between the CTPA and PED map (χ2 =4.60,5.06,P<0.05).The DEPI and PED map had higher consistency in diagnosing PE.The Kappa coefficient was 0.94 if the PED map was regarded as the reference standard,Among 48 cases with segmental PE detected by the PED map,there were 13 cases of complete filling defects and 35 cases of partial filling defects.Among 13 cases of complete filling defects,there were perfusion defect in 10, and perfusion thin in 3 on the DEPI map.Among 35 cases of partial filling defects,there were perfusion defect in 2,perfusion thin in 29,and perfusion normal in 4 on the DEPI map.Conclusion The PED map combined the DEPI map of dual source CT can significantly improve the diagnosis rate of peripheral PE and has a high clinical value.

Objective To observe the effect of pituitary adenylate cyclase activiting polypeptide (PACAP) on traumatic brain injury (TBI) and on tbeCD4+/CD8+ T cell number in blood and spleen of rats.MethodsThe male SD rats were randomly (random number) divided into sham operation group ( n =6),normal saline + TBI group ( n =6) and PACAP + TBI group ( n =6).Right parietal cortical contusion was produced by a weight-dropping method.PACAP was administered intra-cerebroventricularly in a dose of 1 μg/5 μl 20 min before TBI.Brain tissue samples were taken 24 h after trauma.The injured brain tissue of rats was examined by HE stain.The numbers of CD4+/CD8+ T cells in blood and spleen were deteced with flow cytometry.Results Edema,hemorrhage,inflammatory cell infiltration,swollen,degenerated neurons and neurons arrayed disorderly around the injured cortex in hippocampus were found under light microscope.The average numbers of CD4 + T lymphocytes counts in blood and spleen were lower ( P =0.000,P =0.005 ),and number of CD8 + T lymphocytes was higher ( P =0.01 ) in TBI rats group than those in the sham operation group.Micro-injection of PACAP into cerebroventricular attenuated the injury,significantly increased the number of CD4 + T lymphocytes in blood and spleen ( P =0.019,P =0.839),and decreased the number of CD8 + T lymphocytes.ConclusionsPretreatment with PACAP may protect against TBI via influencing periphery T cellular immune function.

Objective To examine the effect of voice cues on nonfluent aphasia patients'performance in reading idioms.Methods Very familiar idioms were presented to aphasia patients on a computer screen either with or without an accompanying voice cue.The patients were asked to say the last two words of each idiom.The number of errors with the last two words of the idioms wag used to compare the patients'performance under the two conditions.Results The mean correct score was 9.46±13.08 when the idioms were presented without a voice cue,but 13.61±6.06 with the voice cue.The difference was statistically significant.Conclusion Voice cues can improve the speaking of familiar idioms for nonfluent aphasia patients.

Keeping an enzyme in its native form with high catalytic activity is of great significance. In the present study, thermal stabilizers of horseradish peroxidase (HRP) were screened. The results indicated that thermal stability of HRP was enhanced by magnesium sulphate and gelatin. A synergic effect of magnesium sulphate and gelatin was observed. In the presence of the stabilizer, the enzymatic activity of HRP remained 89% after kept for 80 h at 50 degrees C and 57% for 90 days at room temperature. Thermal alterations of HRP structure in the absence and presence of the stabilizers were explored by using UV absorption spectra at 402 nm (Soret band), intrinsic fluorescence and 8-anilinonaphthalene-1-sulfonic acid (ANS) fluorescence. The results suggested that magnesium sulphate and gelatin attenuated the extent of unfolding of HRP and therefore the native enzyme structure was stabilized.
Drug Synergism ; Enzyme Stability ; drug effects ; Gelatin ; pharmacology ; Horseradish Peroxidase ; metabolism ; Hot Temperature ; Magnesium Sulfate ; pharmacology

OBJECTIVETo evaluate the efficacy and safety of the HAA regimen (homoharringtonine,cytarabine and aclarubicin)as salvage chemotherapy in the treatment of refractory/relapsed acute myeloid leukemia (AML).

METHODSWe retrospectively analyzed 64 patients with refractory/relapsed AML who received the HAA regimen as salvage chemotherapy. The complete remission (CR)rate was analyzed. Kaplan-Meier method was used to estimate overall survival (OS) and relapse free survival (RFS), and the differences were compared by Log-rank test.

RESULTSThe overall CR rate was 70.1%, and 67.1% of the patients attained CR after the first induction course. The early death rate was 0. The median follow-up time was 61 (range:6-120) months. The estimated 3-year OS rate was 46.8% and the estimated 3-year RFS rate was 42.8%. The CR rates of patients with favorable/intermediate and unfavorable cytogenetics were 76.4% and 33.3%, respectively. The 3-year OS of favorable/intermediate and unfavorable group were 53.7% and 10.0%, respectively. The median survival time of unfavorable group was only 8 months. The side effects associated with the HAA regimen were tolerable, in which the most common toxicities were myelosuppression and infection.

CONCLUSIONHAA regimen is associated with a higher rate of CR and longer-term survival and its toxicity can be tolerated. The regimen is suitable for refractory/relapsed AML patients with favorable or intermediate risk .

Aclarubicin ; analogs & derivatives ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; Cytarabine ; therapeutic use ; Harringtonines ; therapeutic use ; Humans ; Leukemia, Myeloid, Acute ; drug therapy ; Recurrence ; Remission Induction ; Retrospective Studies ; Salvage Therapy ; Survival Rate

Objective To investigate the effect of apatinib on the migration ability of nasopharyngeal carcinoma NPC cells after X-ray irradiation and involved protein expressions. Methods The migration abilities of human immortalized nasopharyngeal epithelial cells ( NP69) and nasopharyngeal carcinoma cells ( CNE-1, CNE-2 ) treated with different concentrations of apatinib ( 0, 5, 10 and 15 μmol/L) were compared by wound healing assay. The effect of apatinib on the activity of NPC cells was detected by CCK-8 for determining the suitable intervention concentration of apatinib. Then NPC cells were divided into control group, apatinib group (15 μmol/L), X-ray irradiation group and apatinib combined with X-ray irradiation group, and the migration ability of each group was compared by wound healing assay. The expressions of pVEGFR2, pSTAT3, STAT3, MMP-9 and EMT related proteins were detected by western blot. Results Compared with the NP69, the migration abilities of CNE-1 and CNE-2 were significantly enhanced ( t=-5. 759, -16. 578, P<0. 05) . Compared with the control group ( 0 μmol/L) , the migration ability of NPC cells after treatment with apatinib(5, 10 and 15 μmol/L) was significantly decreased in a concentration dependent manner ( t=2. 804-13. 362, P<0. 05) . Compared with the X-ray irradiation group, the wound healing rate of NPC cells in the apatinib combined with X-ray irradiation group was decreased ( t=5. 932, 2. 791, P<0. 05) , indicating that apatinib can significantly inhibit the migration of NPC cells after X-ray irradiation. Western blot assay showed that the expressions of pVEGFR2 and pSTAT3 were significantly decreased in NPC cells treated with apatinib, meanwhile, the expression of MMP-9 protein was significantly decreased, and the EMT-related protein was changed. Conclusions Apatinib inhibits migration of X-ray irradiated NPC cells by inhibiting EMT through down-regulating VEGFR2/STAT3/MMP-9 signaling pathway.