ObjectiveTo explore the effects of heat treatment and ultraviolet B (UVB) radiation alone or in combination on the expression of heat shock protein (HSP) 72 in human epidermal melanocytes.Methods Melanocytes were obtained from human foreskin,and subjected to primary culture.After 3 to 5 passages,the melanocytes were classified into 4 groups:control group (receiving no treatment),heat treatment group (treated with heat at 42 ℃ for 1 hour every day for 3 days),UVB group(irradiated with UVB at 50 mJ/cm2 daily for 3days),combination group(treated with heat at 42 ℃ for 1 hour followed by irradiation with UVB at 50 mJ/cm2daily for 3 days).After another 2- to 6-hour culture following the last treatment,melanocytes were collected and subjected to real time PCR and Western blot for the detection of HSP72 mRNA and protein expression,respectively.ResultsThe mRNA and protein expressions of HSP72 were significantly higher in the heat treatment group and combination group than in the control group (mRNA:6.584 ± 0.871 and 7.269 ± 0.454 vs.0.975 ± 0.089,both P ＜ 0.001; protein:2.022 ± 0.058 and 2.080 ± 0.045 vs.0.532 ± 0.033,both P ＜ 0.001 ),but was similar between the UVB group and control group (mRNA:0.832 ± 0.084 vs.0.975 ± 0.089,P ＞ 0.05;protein:0.546±0.021 vs.0.532 ± 0.033,P ＞ 0.05).The ANOVA of factorial design showed that neither heat treatment nor UVB irradiation had interaction effect on the mRNA or protein expression of HSP72 (F =2.106,1.399 respectively,both P ＜ 0.05).ConclusionsHeat treatment can cause an increase in the expression of HSP72,which may enhance the function of melanocytes and protect melanocytes from UVB induced damage.

ObjectiveTo investigate the effect of heat treatment combined with narrow band ultraviolet B(NB-UVB) on cultured normal human melanocytes in vitro.MethodsMelanocytes were isolated from the foreskin of normal human,cullured in vitro,and irradiated with NB-UVB of different doses(20,30,50,70,90,120 and 180 mJ/cm2).Then,MTT assay was performed to evaluate the proliferation and activity of melanocytes to determine the optimal dose of UVB for the next experiment.Melanocytes were classified into 3 groups to be treated with heat at 42 ℃ for 1 hour (heat group),irradiated with UVB at 50 mJ/cm2 (UVB group),or irradiated with UVB at 50 mJ/cm2 followed by heat treatment at 42 ℃ for 1 hour (combination group),daily for 3 successive days; those receiving no treatment served as the control.After 24-hour culture following the last treatment,tyrosinase activity was evaluated with L-dopa as the substrate,melanin content was detected by NaOH assay,and cell cycle stages were determined by flow cytometry.ResultsNB-UVB irradiation decreased the viability of melanocytes in a dose-dependent manner,and the optimum dose of UVB was 50 mJ/cm2.The tyrosinase activity of melanocytes was 0.244 ± 0.018 and 0.310 ± 0.015 respectively in the UVB group and combination group,and increased by 3.8％ (P ＜ 0.05) and 31.9％ (P ＜ 0.05) respectively compared with the control group (0.235 ± 0.018); the melanin content was 0.201 ± 0.016 and 0.286 ± 0.019,respectively in the UVB group and combination group,and increased by 17.5％ (P ＜ 0.05 ) and 67.3％ (P ＜ 0.05) compared with the control group (0.171 ± 0.016).In comparison with the control group,the percentage of melanocytes in G1 phase was decreased by 23.94％ in the UVB group(P＜ 0.05) and 33.51％ in the combination group(P ＜ 0.05),while that in S phase and G2 phase increased by 15.35％ (P ＜ 0.05 ) and 11.93％ (P ＜ 0.05),respectively in the UVB group,and 17.76％ (P ＞ 0.05) and 16.08％ (P ＞ 0.05),respectively in the heat group.ConclusionHeat treatment and NB-UVB can synergistically enhance the tyrosinase activity and accelerate melanogenesis,proliferation and differentiation,of melanocytes.

ObjectiveTo observe the increasing effect of infrared irradiation on tyrosinase activity and melanin content in cultured normal human epidermal melanocytes in vitro and to explore the optimal dose of infrared irradiation.MethodsEpidermal melanocytes were isolated from normal human foreskin tissue,and subjected to primary culture.Methyl thiazolyl tetrazolium(MTT) assay was performed to evaluate the effect of different doses(0,20,60,80,100,140,240,320 J/cm2)of infrared light on the proliferation of melanocytes and to select the optimal irradiation dose.Then,melanocytes were irradiated with infrared light at the optimal dose for 3 consecutive days followed by the determination of tyrosinase activity,melanin content,and cell cycle via dopa oxidation assay,NaOH solubilization method and flow cytometry,respectively.ResultsThe best intervention dose of infrared light was 80 J/cm2.The tyrosinase activity(A492 nm) and melanin content(A492 nm)were 0.3601 ± 0.0301 and 0.2748 ± 0.0243 respectively in melanocytes after irradiation with infrared light of 80 J/cm2 for 3 days,significantly higher than those in unirradiated melanocytes(0.3114 ± 0.0341,0.2325 ±0.0254,respectively,both P ＜ 0.05),with an increase rate of 15.64％ and 18.19％ respectively.Cell cycle analysis revealed a decline in cell percentage in G1 phase(P ＜ 0.01 ) but a concomitant increase in cell percentage in G2 and S phase (both P ＜ 0.05) in irradiated melanocytes compared with unirradiated melanocytes.ConclusionsThe optimal dose of infrared light is 80 J/cm2 for the irradiation of melanocytes,and this dose of infrared light can increase melanin content,tyrosinase activity,differentiation and proliferation of melanocytes.

Objective To explore the effects of WeChat-based peer supports upon medication adherence and quality-of-life in liver transplant recipients .Methods A total of 63 patients with liver transplantation were conveniently divided into intervention group (n=32) and control group (n=31) depending upon their different follow-up periods .In control group ,routine outpatient health guidance was offered while intervention group received 6-week WeChat-based peer supports . Medication compliance and quality-of-life of two groups were evaluated at Month 3/6/12 post-intervention . Results At Month 3 post-intervention ,as compared with control group ,only non-punctual medication improved significantly in intervention group (P<0 .01);at Month 6 post-intervention ,all aspects of drug adherence improved in intervention group (P< 0 .05) ,at Month 12 post-intervention ,drug adherence ,non-punctual medication and missed dosing improved in intervention group ( P> 0 .05 ) . However , inter-group quality-of-life was not statistically significant at Month 3/6/12 post-intervention .Conclusions WeChat-based peer supports may partially improve the immediate compliance of patients with liver transplantation .However ,long-term outcomes and effects on quality-of-life are worth further researches .

Objective To analyze the clinicopathological features and prognosis of immunoglobulin G (IgG)4-related diseases with interstitial nephritis. Methods Forty cases of IgG4-related diseases diagnosed by pathology in our hospital from 2014 to 2018 were collected and their clinicopathological features were analyzed. Four patients with IgG4-related disease with interstitial nephritis were analyzed, including clinical laboratory tests and histopathological features, and immunohistochemical analysis of the type and proportion of renal interstitial infiltrating cells. At the same time, the treatment and prognosis of the patients were analyzed. Results Among 40 cases of IgG4-related diseases, 11 cases had parotid submandibular gland involvement (accounted for 28％), labial gland involvement (7 cases, 18％) and lymph node enlargement (6 cases, 15％). Patients with kidney involvement (4 cases, 10％) all presented with IgG4-related tubulo-interstitial nephritis (IgG4-TIN). All of them were elderly males, and 2 had glomerulopathy. One of them had anti-neutrophil cytoplasmic antibodies (ANCA)-related vasculitis renal damage. The number of CD4-positive cells in renal interstitium was more than CD8-positive cells. Another case complicated with IgA nephropathy. Renal dysfunction occurred in all 4 cases, and serum IgG4 level ranged from 4.65 g/L to 23.8 g/L. All 4 patients received glucocorticosteroid and symptomatic treatment, and the prognosis was good. Conclusion IgG4-related diseases may involve mul-tiple organs, renal dysfunction may occur when the kidney is involved. Interstitial nephritis is the major clinical manifestation. Glomerular lesions may accelerate the progress of IgG4-TIN. Corticosteroid therapy is effective.

Arabidopsis AtPRMT10 is a plant-specific type I protein arginine methyltransferase that can asymmetrically dimethylate arginine 3 of histone H4 with auto-methylation activity. Mutations of AtPRMT10 derepress FLOWERING LOCUS C (FLC) expression resulting in a late-flowering phenotype. Here, to further investigate the biochemical characteristics of AtPRMT10, we analyzed a series of mutated forms of the AtPRMT10 protein. We demonstrate that the conserved "VLD" residues and "double-E loop" are essential for enzymatic activity of AtPRMT10. In addition, we show that Arg54 and Cys259 of AtPRMT10, two residues unreported in animals, are also important for its enzymatic activity. We find that Arg13 of AtPRMT10 is the auto-methylation site. However, substitution of Arg13 to Lys13 does not affect its enzymatic activity. In vivo complementation assays reveal that plants expressing AtPRMT10 with VLD-AAA, E143Q or E152Q mutations retain high levels of FLC expression and fail to rescue the late-flowering phenotype of atprmt10 plants. Taken together, we conclude that the methyltransferase activity of AtPRMT10 is essential for repressing FLC expression and promoting flowering in Arabidopsis.
Arabidopsis ; enzymology ; Arabidopsis Proteins ; biosynthesis ; genetics ; metabolism ; Enzyme Activation ; Flowers ; genetics ; growth & development ; metabolism ; Genetic Loci ; genetics ; MADS Domain Proteins ; biosynthesis ; genetics ; metabolism ; Methyltransferases ; genetics ; metabolism ; Phenotype ; Recombinant Proteins ; genetics ; metabolism ; Time Factors

Genome editing is a novel targeted genome modification biotechnology, which could successfully mutate specific loci as well as generate gene replacement and insertion in various organisms. So far, genome editing technology has been widely applied in investigating gene function and developing valuable traits in both model plants and major crops. In this review, we briefly survey the historical development of genome editing technology, summarize recent progress using the CRISPR/Cas9 system for plant genome editing and explore the potential of the CRISPR/Cas technology in improving forage legumes.

Objective:To investigate the efficacy of concurrent chemoradiotherapy combined with icotinib targeted therapy for patients with epidermal growth factor receptor (EGFR)-mutated non-small cell lung cancer (NSCLC).Methods:A total of 89 EGFR-mutated NSCLC patients who were admitted to Shanxi Province Cancer Hospital from January 2017 to January 2019 were selected and divided into control group (45 cases) and observation group (44 cases) by random number table method. The control group received cisplatin combined with docetaxel concurrent chemoradiotherapy, the observation group received cisplatin combined with docetaxel concurrent chemoradiotherapy and oral icotinib targeted therapy. The blood coagulation function, immune function and levels of tumor markers were compared between the two groups.Results:There was no statistical difference in blood coagulation function, immune function and levels of tumor markers between the two groups before treatment (all P > 0.05). After treatment, the levels of fibrinogen [(13±4) g/L vs. (16±6) g/L], D-dimer [(1.0±0.8) mg/L vs. (1.4±1.0) mg/L], squamous cell carcinoma antigen [(0.97±0.23) μg/L vs. (1.11±0.21) μg/L], carbohydrate antigen 125 [(21±7) U/ml vs. (35±11) U/ml] and carcinoembryonic antigen [(2.2±0.3) ng/ml vs. (6.0±1.1) ng/ml] in the observation group were lower than those in the control group, and the differences were statistically significant ( t values were 2.84, 2.11, 3.08, 7.40 and 23.08, all P < 0.05). After treatment, the ratios of NK cells [(18±7)% vs. (15±4)%], cytotoxic T cells [(17.2±6.1)% vs. (14.7±3.6)%] and helper T cells [(31.03±0.11)% to (25.88±0.39)%] in the observation group were higher than those in the control group, and the differences were statistically significant ( t values were -2.91, -2.59 and 2.79, all P < 0.05). Conclusions:Concurrent chemoradiotherapy combined with icotinib targeted therapy can better improve the hypercoagulable state and levels of tumor markers in patients with EGFR-mutated NSCLC than simple concurrent chemoradiotherapy, and can improve the immune function of patients, which has good therapeutic efficacy.

Objective:To analyze the distribution and drug resistance of wound pathogenic microorganisms in outpatients of wound healing center so as to provide a basis for the standardized construction of wound healing centers.Methods:A retrospective case series study was used to analyzed the data of 365 outpatients treated at Ruijin Hospital, Shanghai Jiaotong University School of Medicine from December 2017 to October 2019. There were 220 males and 145 females, aged (58.8±18.9)years (range, 18-98 years). The patients included 92 first-visit patients and 273 re-visit patients. The culture results (positive rate of pathogenic microorganisms, bacterial species, bacterial distribution) and drug sensitivity results of the wound secretions were compared and analyzed.Results:(1) Among 365 samples of wound secretions, 198 patients were positive for pathogenic microorganisms with a positive rate of 54.3%. A total of 107 strains (51.0%) of Gram-positive bacteria were detected, mainly Staphylococcus aureus (70 strains, 33.3%); 95 strains (45.2%) of Gram-negative bacteria were detected, mainly Escherichia coli (20 strains, 9.5%), followed by Pseudomonas aeruginosa (17 strains, 8.1%); 8 strains (3.8%) of fungi were detected. (2) A total of 26 (28.3%) first-visit patients were positive for pathogenic microorganisms, and 172 (63.0%) re-visit patients were positive for pathogenic microorganisms. The rate of positive microorganism detection had significant differences between first-visit and re-visit patients ( P<0.05). (3) A total of 29 strains were detected in first-visit patients, including 16 strains (55.2%) of Gram-positive bacteria, 11 strains (37.9%) of Gram-negative bacteria and 2 strains (6.9%) of fungi. A total of 181 strains were detected in re-visit patients, including 91 strains (50.3%) of Gram-positive bacteria, 84 strains (46.4%) of Gram-negative bacteria and 6 strains (3.3%) of fungi. The microbial distribution was significantly different between first-visit and re-visit patients ( P<0.05). (4) Compared with first-visit patients, the resistance of Staphylococcus aureus isolated from the re-visit patients to spenicillin, oxacillin, ciprofloxacin, tetracycline, clindamycin, moxifloxacin, erythromycin, and levofloxacin were increased variably. No vancomycin-resistant Staphylococcus aureus was detected, indicating that the staphylococcus aureus presented in the wound was highly sensitive to vancomycin. Conclusions:Staphylococcus aureus is the most common microorganism in wound secretions in outpatients of wound healing center. The rate of positive pathogenic microorganisms in wound secretions of re-visit patients is significantly higher than that of first-visit patients, and the distribution of pathogenic microorganisms of first-visited and revisited patients differs significantly. The Staphylococcus aureus detected in re-visit patients has a higher resistance to common antibiotics compared with first-visit patients. It is suggested that timely detection of pathogenic microorganisms in outpatients and effective control and supervision of outpatient infections are important contents that cannot be ignored in the construction of wound healing center.

ObjectiveTo explore the association between short-term exposure to atmospheric fine particulate matter （PM_2.5） and ozone （O₃） and systemic inflammatory indicators in patients with pneumonia， and to identify the susceptible populations. MethodsFrom September 2018 to April 2020， data of 1 480 patients admitted for pneumonia was collected from a tertiary hospital in Taiyuan City. Generalized additive models （GAMs） were used to explore the associations between PM_2.5 and O₃ exposure and inflammatory indicators of patients with pneumonia； and to explore the susceptibility factors and susceptible populations to PM_2.5 and O₃ exposures through stratified analyses. ResultsThe short-term exposure to PM_2.5 was associated with changes in peripheral blood C-reation protein （CRP）， erythrocyte sedimentation （ESR）， easinophil （EOS）， neutrophil （NEU） and neutrophil-lymphocyte ratio （NLR） in patients with pneumonia， and there were different degrees of hysteresis effects， with the effect values reaching a maximum at lag03， lag03， lag0， lag03， lag03， respectively， which were 4.13% （95%CI： 0.43%‒7.84%）， 3.10% （95%CI： 0.24%‒5.97%）， 5.27% （95%CI： 3.12%‒7.42%）， 1.85% （95%CI： 0.36%‒3.34%）， and 2.53% （95%CI： 0.53%‒4.74%） for every 10 μg·m^-3 of PM_2.5. The changes in O₃ concentration were associated with the elevation of peripheral blood PCT and ESR in patients with pneumonia， and their effect values all reached the maximum at lag01 d， every 1 μg·m^-3 of O₃ elevation increased by 0.38% （95%CI： 0.04%‒0.73%） and 0.47% （95%CI： 0.19%‒0.76%）， respectively. Stratified analyses showed that the associations of PM_2.5 with peripheral blood CRP， ESR， NEU， and NLR in pneumonia patients were more significant in males， the elderly， and those with onset in the cold season； the associations of O₃ with peripheral blood PCT and ESR in pneumonia patients were more significant in the elderly and those with onset in the warm season， and the peripheral blood CRP and PCT in female patients with pneumonia were more susceptible to the changes of O₃. ConclusionShort-term exposure to atmospheric PM_2.5 and O₃ are positively associated with changes in inflammatory indicators in patients with pneumonia， and the effects of PM_2.5 on patients with pneumonia are more extensive than those of O₃， with a longer lag effect. In addition， elderly patients with pneumonia are more sensitive to air pollution， male patients with pneumonia are more sensitive to PM_2.5， and female patients with pneumonia are more sensitive to O₃. Cold and warm seasons can exacerbate the effects of PM_2.5 and O₃ on inflammatory indicators in patients with pneumonia， respectively， and the patients must be protected well.