ABSTRACT
Objective:To observe the clinical efficacy of micro-needle therapy combined with Biotrisse BTS Lumin OX and Biotrisse BTS Brightline in the treatment of melasma.Methods:From September 2019 to June 2021, 80 patients with facial chloasma, aged 28-48 (37.3±4.9) years, were selected from Nanjing Jiangning Guze Clinic. The micro-needle therapy was combined with lumin OX and brightline for 6 times, and the observation time was 120 days. The mMASI score and VISIA photos before and after treatment were used to improve the results.Results:Eighty patients with refractory melasma on the face were treated with micro-needle therapy for 7 times (1 time per week for the first 5 times, and twice a week for the 9th and 10th weeks). The photos before and after treatment were compared and the mMASI of the patients' facial chloasma was compared. The scores were analyzed statistically, and the melasma were improved to varying degrees. Before treatment, the mMASI score was 5.4±3.22; 90 days later, the mMASI was 3.22±2.16, and the score decreased significantly ( t=5.9, P<0.05); after 120 days, mMASI score was 1.6±0.68, and the score decreased significantly ( t=7.55, P<0.05). Pigmentation occurred in one patient after treatment, and hypopigmentation after repair treatment; none of the patients had adverse reactions such as hypopigmentation. All the 80 patients had different degrees of improvement in pores and skin texture. Conclusions:The combination of micro-needling with lumin OX and brightline in the treatment of chloasma has a definite effect without obvious side effects. It provides a new method for the treatment of chloasma.
ABSTRACT
Objective:To determine the protective effect of fasudil on acute lung injury in septic mice.Methods:Forty-five 4-6-week-old male C57BL mice were randomly(random number) assigned to three groups ( n=15 each group): control group, lipopolysaccharide (LPS) group and Fasudil intervention group (FAS+LPS). Acute lung injury model of septic mice was established with an intraperitoneal injection and intratracheal infusion of LPS. The mice in the FAS+LPS group were injected with fasudil hydrochloride intraperitoneally 30 min before intraperitoneal LPS injection and 1 h after intratracheal LPS infusion, respectively. All mice were sacrificed at 4 h after modeling, and lung tissues were collected. Hematoxylin-eosin staining was preformed to observe the morphological changes in the lung tissue. The wet /dry weight (W/D) ratio, malondialdehyde (MDA) content and the activity of myeloperoxidase (MPO) in the lung tissues were detected. Caspase-3 expression was examined by immunohistochemical (IHC) staining. Western blot was employed to detect the expression of RhoA, ROCK1, endothelial nitric oxide synthase (eNOS), and p-eNOS. Results:Inflammatory cell infiltration and erythrocyte exudation were significantly reduced, and the degree of interstitial oedema and derangement of alveolar structure appeared in a decreasing degree after FAS intervention. Compared with the LPS group, the W/D ratio, MDA content, MPO activity and the expression of Caspase-3 in the FAS+LPS group were significantly reduced (all P<0.01). Meanwhile, the expression of RhoA and ROCK1 of the LPS group were obviously higher than those in the control group ( P<0.05), and p-eNOS was obviously lower than that in the control group ( P<0.05). Furthermore, the expression of RhoA and ROCK1 of the FAS+LPS group were obviously lower than those in the LPS group, and p-eNOS was obviously higher than that in the LPS group. There was no significant difference on the expression of eNOS among the three groups. Conclusions:Fasudil can alleviate the degree of inflammatory cell infiltration, reduce apoptosis in lung tissue, inhibit the RhoA/ROCK1 signaling activity, and promote the phosphorylation expression of eNOS in septic mice.
ABSTRACT
Objective:To investigate the clinical characteristics and pathogen spectrum of acquired immunodeficiency syndrome (AIDS) patients complicated with pulmonary filamentous fungal infection in Guangdong Province, so as to provide evidences for improving the diagnosis and treatment.Methods:A total of 143 AIDS patients with pulmonary filamentous fungal infection hospitalized in Guangzhou Eighth People′s Hospital, Guangzhou Medical University from January 2016 to December 2018 were included. The filamentous fungi cultured in bronchoalveolar lavage fluid of these patients were identified with morphological and molecular biological methods. And their clinical characteristics were analyzed. Nonparametric Kruskal-Wallis H test and chi-square test were used for statistical analysis. Results:Among the 143 patients, 116(81.1%) had fever, 104(72.7%) had cough, 83(58.0%) had expectoration, and 59(41.3%) had anhelation. The CD4 + T lymphocyte count was 22.0(9.3, 60.8) cells/μL and 118(82.5%) cases were below 100.0 cells/μL. The white blood cell counts decreased in 52(36.4%) cases and increased in 18(12.6%) cases, anemia was found in 109(76.2%) cases, platelet count decreased in 29(20.3%) cases. Sixty-four (44.8%) cases were positive for galactomannan test. Chest computed tomography showed diffuse infection of both lungs in 114(79.7%) cases, miliary changes in 12(8.4%) cases, pleural effusion in 44(30.8%) cases, and enlargement of pleural and (or) mediastinal lymph nodes in 45(31.5%) cases. After receiving antifungal therapy, 124 (86.7%) cases were cured or improved, and 19 (13.3%) cases were discharged automatically or died of disease deterioration. Among the 143 strains of filamentous fungi, there were 56 strains of Aspergillus species pluralis (39.2%, including 24 strains of Aspergillus fumigatus), 37 strains of Talaromyces marneffei ( T. marneffei) (25.9%), 22 strains of Penicilium species pluralis (15.4%), and 28 strains of other genera of filamentous fungi (19.6%). The median CD4 + T lymphocyte counts in patients infected with Aspergillus species pluralis, T. marneffei, Penicilium species pluralis and other genera were 24.5, 15.0, 53.5 and 22.0 cells/μL, respectively, and the difference was statistically significant ( H=11.282, P=0.010). The proportions of AIDS patients with different pulmonary filamentous fungal infection of CD4 + T lymphocyte count ≤50.0 cells/μL in descending order were T. marneffei group (89.2%(33/37)), Aspergillus species pluralis group and other genera group (67.9%(38/56), 67.9%(19/28)), and Penicillium species pluralis group (54.5%(12/22)), and the difference was statistically significant ( χ2=9.296, P=0.026). Conclusions:The clinical manifestations of pulmonary filamentous fungal infection in AIDS patients in Guangdong Province are not specific. The pathogenic spectrum contains various genera, and T. marneffei and Aspergillus fumigatus are dominant, which could be correlated with CD4 + T lymphocyte count.
ABSTRACT
Objective To study the effects VEGF small interfering RNA (siRNA) on chemosinsitivity of human BxPC3 cell and its mechanism. Methods BxPC3 cells were divided into single BxPC3 cell group,lipofection group, scrambled siRNA transfection (200 nmol/L) group, VEGF siRNA transfection group.VEGF siRNA (5, 10, 20, 100,200 nmol/L) was used to transfect BxPC3 cells. Expressions of VEGF mRNA and protein were determined by real-time PGR and ELISA assay, respectively. MTT was performed to examine the inhibitory effects of gemcitabine on BxPC3 cells of each group. The phosphorylated-Akt protein was evaluated by Western blotting. Results After VEGF siRNA transfection, the expression of VEGF mRNA and protein in BxPC3 cells was down-regulated in a dose-and time-dependent manner, but there was no effect on BxPC3 cells proliferation. After 0. 2 μmol/L of gemcitabine treatment for 48 h, the inhibitory rates were ( 16.9 ±0.3)%, (17.3 ±0.3)%, (28.8 ±0.4)%, (52.2 ±0.3)%, (75.4 ±0.4)% in BxPC3 cell group,lipofection group, 5,10,20 nmol/L VEGF siRNA transfection group, and the inhibitory effects were correlated with siRNA concentration ( r = 0. 928 ). The phosphorylated-Akt protein was reduced significantly in siRNA transfected cells. Conclusions VEGF gene plays an important role in BxPC3 cells resistence to chemotherapy through inhibiting Akt phosphorylation.
ABSTRACT
Objective To study the effects of midkine(MK)gene small interfering RNA(siRNA)on adhesion and invasion of human breast cancer cells.Methods Real time PCR was used to evaluate MK mRNA expression in 7 human breast cancer cell lines Bcap-37,LCCI,MCF-7,MDA-MB-231,MDA-MB-435,MDA-MB-468,and ZR75-1.The cell line in which MK expression was the highest was transfected with different doses of MK siRNA.The expression of MK mRNA and protein was determined by real-time quantitative PCR and immunoflurescence staining.The cell adhesion was evaluated by MTT assay and invasion was examined by Boyden chamber method.Results Cell line MCF-7 expressed the highestlevel of MK mRNA in the 7 tested breast cancer cell lines.After being transfected with MK siRNA,MK mRNA and protein level of MCF-7 decreased in timeand dose-dependent manners.The adhesive and invasive ability of MCF-7 cell transfected with MK siRNA decreased in a dose dependent manner(P<0.01,P<0.01).Conclusions MK gene might play an important role in adhesion and invasion of human breast cancer cells.siRNA transfection could effectively inhibit adhesion,migration,and invasion of human breast cancer cell.
ABSTRACT
Objective To study effects of polo-like kinase-1(PLK1)small interfering RNA(siRNA)on proliferation and apoptosis of undifferentiated human thyroid cancer cells.Methods 5 PLK1 siRNA(S1,S2,S3,S4 and S5)were constructed and used to transfect human thyroid cancer cell line ARO.RT-PCR was employed to pick out the most effective siRNA,which was then used to transfect ARO cell.RT-PCR and western blot were used to detect PLK1 expression in thyroid cancer cells,which were divided into different groups.MTT assay was performed to examine the effects of PLK1 siRNA on thyroid cancer cells in all groups.Apoptosis of thyroid cancer cells was observed by caspase-3 activity and TUNEL.Results All the 5 siRNA down-regulated PLK1 mRNA expression.among which S4 showed the best effect.S4 transfection could obviously inhibit proliferation of thyroid cancer cells in dose and time dependent manner.Compared with control groups,caspase-3 activity of cancer cells in s4 transfeeted group increased significantly.The effect of S4 transfection was dose and time dependent.TUNEL results showed apoptosis of cancer cells transfected by S4 siRNA was obvious and apoptosis of cells was dose-dependent.Conclusions PLK1 may play an important role in proliferation of undifferentiated thyroid carcinoma.PLK1 siRNA transfection can inhibit proliferation of throid cancer cell through apoptosis induction.
ABSTRACT
ObjectiveTo study the effects of tumor-associated calcium signal transducer-2 (TROP-2) gene small interfering RNA(siRNA) on adhesion and invasion of human breast cancer cell. MethodsReal time PCR was used to evaluate the TROP-2 mRNA of seven human breast cancer cell lines Bcap-37 ,LCC1 ,MCF-7 ,MDA-MB-231,MDA-MB-435, MDA-MB-468 ,and ZR75-1. The cell line of TROP-2 highest expression was transfected with different dose of TROP-2 siRNA. The expression of TROP-2 mRNA and protein were determined by Real-time quantitative PCR and immumoflurescence method. The cell adhesion was evaluated by MTT assay,and invasion was exmined by hoyden chamber,respectively. Results Cell line MCF-7 showed the highest elevation of TROP-2 mRNA in seven breast cancer cell lines. The results from real-time quantitative PCR and immumoflurescence method showed that TROP-2 mRNA and protein reduced in time-and dose-dependent manners( P ＜ 0.01 ;P ＜ 0.01 ). The adhesive rate of siRNA groups(5 nM,10 nM,and 20 nM)was(52.9 +2.5)％ ,(25.6 ±2.3)％, ( 12.8 +2.2)％ (P ＜0.01 ) ,respectively.The transwell results showed that the invasion cells was(78 ± 17), (39 ± 15), ( 19 ± 16), ( 136 +25 ) and( 139 ±21 )in different groups(5,10,20 nM siRNA,and controls) ,respectively(P ＜0.01). ConclusionTROP-2 gene might play an important role in adhesion and invasion of human breast cancer cell. siRNA targeted TROP-2 could effectively inhibit adhesion and invasion of human breast cancer cell.
ABSTRACT
Objective To investigate the effects of S100A6 gene on invasion of human pancreatic cancer cell and possible mechanism. Methods Human pancreatic cancer BxPC3 cell line was transfected with small interfering RNA (siRNA) targeting S1006 gene, the mRNA and protein levels of S100A6 were determined by real time RT-PCR and Western blotting respectively. The invasion ability was evaluated by Transwell chamber. The matrix metalloproteinase-2 (MMP-9) activity of cancer cells was examined by gelatin zymography. Results The levels of mRNA and protein of S100A6 were greatly reduced in a dose and time dependent manner, the number of penetrating cells was greatly reduced in a dose dependent manner. The expression of S100A6 mRNA in 12.5 nmol/L of S100A6 siRNA transfected group decreased from ( 100 ±0.3)％ in control group to (15.3 ±0.2)％ ; while the expression of S100A6 protein decreased from (83.2 ±0. 18 ) ％ to ( 13.5 ± 0. 12) ％ ; the number of penetrating cells decreased from 44.5 ± 2.2 to 7.6 + 1.5 ( P ＜0. 01 ). The MMP-9 activity of siRNA group reduced significantly. Conclusions S100A6 siRNA can inhibit the invasion of pancreatic cancer cells through down-regulation of MMP-9.
ABSTRACT
Objective To study the effects of midkine (MK) gene-targeting small interfering RNA (siRNA)on the invasion of melanoma cells.Methods Three MK gene-targeting siRNAs (S1,S2 and S3)were designed,constructed,and transfected into human A375 melanoma cells.Real-time PCR was performed to measure the expression of MK gene and to screen the siRNA with best efficacy.Then,A375 cells were transfected with the optimal siRNA of various doses (3.125,6.25 and 12.5 nmol/L)followed by additional culture of various durations(24,48,72 hours).Some A375 cells remaining untreated served as the blank control group,and some transfected only with liposomes served as the vector control group.Reverse transcription (RT) -PCR and Western blot were conducted to detect the mRNA and protein expression of MK,respectively,MTT assay to observe the adhesion of A375 cells,and Boyden chamber was used to evaluate cell invasion.Results The expression of MK mRNA was downregulated by all the three siRNAs,especially by the siRNA S3,which was used in the following transfection experiment.Real-time quantitative PCR revealed that the MK mRNA expression was reduced by the siRNA in a dose- (r24hours=-0.906,r4Bhours=-0.922,r72hours=-0.939,all P<0.01)and time-dependent(r3.125nmol/L=-0.889,r625nmol/L=-0.935,r125nmol/L=-0.928,all P<0.01)manner.MTT assay showed that the percentage of adhesing cells was 73.66%±2.25%,49.36%±2.16%and 28.35%±1.68%in A375 cells transfected with the siRNA of 3.125,6.25 and 12.5 nmol/L,respectively.The number of cells migrating across the chamber filter was 23.9±1.6,12.1±1.5,5.6±1.2 among A375 cells transfected with the siRNA of 3.125,6.25 and 12.5 nmol/L,respectively,significantly lower than that in the blank control group(36.8±1.5).The percentage of adhesing cells and number of migrating cells decreased with the dose of siRNA(r=-0.936,-0.915,P<0.01,0.05,respectively).Conclusions MK gene might play an important role in the adhesion and invasion of melanonla cells.To down-regulate the expression of MK gene by siRNA may suppress the adhesion and invasion of melanoma cells.