Objective To explore the effects of Qingshen Granules(QSG)on adenine-induced renal fibrosis in mice and in uric acid(UA)-stimulated NRK-49F cells and its mechanism for regulating exosomes,miR-330-3p and CREBBP.Methods A mouse model of adenine-induced renal fibrosis were treated daily with QSG at 8.0 g·kg-1·d-1 via gavage for 12 weeks.An adeno-associated virus vector was injected into the tail vein,and renal tissues of the mice were collected for analyzing exosomal marker proteins CD9,Hsp70,and TSG101 and expressions of Col-Ⅲ,α-SMA,FN,and E-cad using Western blotting and immunofluorescence and for observing pathological changes using HE and Masson staining.In the cell experiment,NRK-49F cells were stimulated with uric acid(400 μmol/L)followed by treatment with QSG-medicated serum from SD rats,and the changes in expressions of the exosomal markers and Col-Ⅲ,α-SMA,FN,and E-cad were analyzed.Dual luciferase reporter assay was employed to examine the targeting relationship between miR-330-3p and CREBBP,whose expressions were detected by RT-qPCR and Western blotting in treated NRK-49F cells.Results The mouse models of adenine-induced renal fibrosis showed significantly increased levels of CD9,Hsp70,and TSG101,which were decreased by treatment with QSG.The expressions of Col-Ⅲ,α-SMA,and FN increased and E-cad decreased in the mouse models but these changes were reversed by QSG treatment.QSG treatment obviously alleviated renal fibrosis in the mouse models.Intravenous injection of adeno-associated viral vector obviously inhibited miR-330-3p,increased CREBBP levels,and reduced fibrosis in the mouse models.Dual luciferase assay confirmed CREBBP as a target of miR-330-3p,which was consistent with the results of the cell experiments.Conclusion QSG inhibits renal fibrosis in mice by regulating the exosomes,reducing miR-330-3p levels,and increasing CREBBP expression.

Objective To explore the effects of Qingshen Granules(QSG)on adenine-induced renal fibrosis in mice and in uric acid(UA)-stimulated NRK-49F cells and its mechanism for regulating exosomes,miR-330-3p and CREBBP.Methods A mouse model of adenine-induced renal fibrosis were treated daily with QSG at 8.0 g·kg-1·d-1 via gavage for 12 weeks.An adeno-associated virus vector was injected into the tail vein,and renal tissues of the mice were collected for analyzing exosomal marker proteins CD9,Hsp70,and TSG101 and expressions of Col-Ⅲ,α-SMA,FN,and E-cad using Western blotting and immunofluorescence and for observing pathological changes using HE and Masson staining.In the cell experiment,NRK-49F cells were stimulated with uric acid(400 μmol/L)followed by treatment with QSG-medicated serum from SD rats,and the changes in expressions of the exosomal markers and Col-Ⅲ,α-SMA,FN,and E-cad were analyzed.Dual luciferase reporter assay was employed to examine the targeting relationship between miR-330-3p and CREBBP,whose expressions were detected by RT-qPCR and Western blotting in treated NRK-49F cells.Results The mouse models of adenine-induced renal fibrosis showed significantly increased levels of CD9,Hsp70,and TSG101,which were decreased by treatment with QSG.The expressions of Col-Ⅲ,α-SMA,and FN increased and E-cad decreased in the mouse models but these changes were reversed by QSG treatment.QSG treatment obviously alleviated renal fibrosis in the mouse models.Intravenous injection of adeno-associated viral vector obviously inhibited miR-330-3p,increased CREBBP levels,and reduced fibrosis in the mouse models.Dual luciferase assay confirmed CREBBP as a target of miR-330-3p,which was consistent with the results of the cell experiments.Conclusion QSG inhibits renal fibrosis in mice by regulating the exosomes,reducing miR-330-3p levels,and increasing CREBBP expression.

Objective:To evaluate the efficacy and safety of narrow-band intense pulsed light in the treatment of erythromelanosis follicularis faciei et colli (EFFC).Methods:The patients with EFFC, diagnosed at the Department of Cosmetic Laser Surgery, Hospital for Skin Disease, Chinese Academy of Medical Sciences, and Peking Union Medical College, from January 2017 to December 2022, were retrospectively evaluated. There were 28 patients, including 24 males and 4 females. They ranged in age from 15 to 35 (21.0±4.2) years. All patients received three sessions of narrow-band intense pulsed light treatments, with intervals of 1-2 months. A follow-up visit was completed 1-2 months after three treatments, and efficacy was evaluated. Two dermatologists assessed the clinical improvement in erythema, pigmentation, and follicular papules on a 4-point scale (0=0-25% improvement, 1=26%-50% improvement, 2=51%-75% improvement, and 3=76%-100% improvement). Adverse events were also recorded.Results:A total of 28 patients received three sessions of narrow-band intense pulsed light treatments and were followed-up. After three treatments, the mean improvement scores for erythema, follicular papules, and pigmentation were (2.02±0.65), (1.38±0.59) and (0.80±0.61) respectively. Among these patients, 67.9% (19/28) experienced over 50% improvement in erythema, 39.3% (11/28) showed over 50% improvement in follicular papules, and 7.1% (2/28) demonstrated over 50% improvement in pigmentation. No blisters and scars were formed.Conclusions:Narrow-band intense pulsed light is a safe and effective treatment for patients with EFFC, particularly for those presenting with erythema.

Objective:To investigate the effect of miRNA (miR) -193b-5p on melanogenesis and its possible mechanisms.Methods:Human primary melanocytes were isolated from discarded normal foreskin tissues of healthy males after circumcision, and cultured in vitro. miR-NC mimics (miR-NC mimic group) and miR-193b-5p mimics (miR-193b-5p mimic group) were transfected into human primary melanocytes and human MNT1 melanoma cells, separately. After transfection, real-time quantitative PCR (RT-qPCR) was performed to determine the overexpression efficiency of miR-193b-5p at 48 hours, Western blot analysis to determine the expression of melanogenesis-related proteins tyrosinase (TYR) and microphthalmia-associated transcription factor (MITF) in human primary melanocytes and human MNT1 melanoma cells at 72 hours, and the melanin content in the above cells was determined by a sodium hydroxide solubilization method at 1 week. The target gene of miR-193b-5p was predicted by using Targetscan algorithms and verified by dual-luciferase reporter assay, and RT-qPCR and Western blot analysis were performed to analyze changes in mRNA and protein expression of the target gene respectively after the overexpression of miR-193b-5p. Two-independent-samples t test was used for comparisons between two groups. Results:In human primary melanocytes and human MNT1 melanoma cells, the miR-193b-5p expression levels were significantly higher in the miR-193b-5p mimic groups than in the miR-NC mimic groups ( t = 65.57, 22.49, respectively, both P < 0.001) , and the melanin content was significantly lower in the miR-193b-5p mimic groups (0.091 ± 0.007, 0.130 ± 0.004, respectively) than in the miR-NC mimic groups (0.117 ± 0.002, 0.188 ± 0.032, t = 5.98, 3.24, P < 0.01, < 0.05, respectively) . Western blot analysis showed that the expression of melanogenesis-related proteins TYR and MITF in both human primary melanocytes and human MNT1 melanoma cells was significantly lower in the miR-193b-5p mimic groups than in the miR-NC mimic groups (all P < 0.01) . TargetScan analysis and dual-luciferase reporter assay revealed a binding site for miR-193b-5p in the 3′ untranslated region of the transcriptional regulator CITED2. After up-regulation of miR-193b-5p expression in human primary melanocytes and human MNT1 melanoma cells, the CITED2 mRNA and protein expression levels significantly decreased compared with the miR-NC mimic groups (all P < 0.05) . Conclusion:miR-193b-5p overexpression can down-regulate the expression of melanogenesis-related proteins TYR and MITF, and then inhibit melanogenesis, which may be related to the targeted inhibition of CITED2 expression.

Objective:To construct an artificial intelligence model for the diagnosis of facial vitiligo, so as to realize artificial intelligence-assisted diagnosis of facial vitiligo.Methods:Based on digital single-lens reflex (SLR) camera images of vitiligo skin lesions and YOLO (You Only Look Once) v3 algorithm, a skin lesion detection model Vit3 was established, and its performance was evaluated by comparing its detection results and labeling results of dermatologists. On the basis of the Vit3 model, both optical and ultraviolet images of vitiligo and non-vitiligo skin lesions were taken by using an artificial intelligence-based facial skin image collector, and the gray values of vitiligo and non-vitiligo skin lesion areas on the ultraviolet images were measured by using an image processing technique. According to the gray-value threshold between vitiligo and non-vitiligo skin lesions, a facial vitiligo diagnosis model Vit4 was established. Cochran′s Q test was used to compare the diagnostic results of the Vit4 model and dermatologists, and the diagnostic performance of the Vit4 model was evaluated. Results:For 100 SLR camera images of vitiligo skin lesions (167 lesional sites) and 100 SLR camera images of normal skin, the diagnostic sensitivity of the Vit3 model was 92.81% (155/167) . For 97 pairs of facial skin images (including 50 vitiligo lesions, 30 pityriasis alba lesions, 7 amelanotic nevus leisons, and 10 normal skin tissues) , the diagnostic accuracy rate, sensitivity and specificity of the Vit4 model were 88.66% (86/97) , 88.00% (44/50) and 89.36% (42/47) respectively, and there was no significant difference in the diagnostic accuracy rate between the Vit4 model and dermatologists (92.78%[90/97], χ2=2.323, P > 0.05) . Conclusion:The artificial intelligence model Vit4 was established for the diagnosis of facial vitiligo with favorable diagnostic performance, and could serve as an objective and convenient method for the auxiliary diagnosis of facial vitiligo.

OBJECTIVE: To compare the accuracy of five warfarin-dosing algorithms and warfarin stable dose model (2.5 mg/day) for Shandong population. METHODS: One hundred and twenty five patients who achieved stable warfarin dose were enrolled. Clinical and genetic data were used to evaluate the value of each algorithm by calculating the percentage of patients whose predicted warfarin dose was within 20% of the actual stable therapeutic dose and mean absolute error (MAE). RESULTS: The frequency of patients with CYP2C9*1/*1, CYP2C9*1/*3 and CYP2C9*1/*2 genotype was 92.00%, 7.20%, 0.80%, respectively. That of VKORC1-1639 AA, AG and GG genotype was 82.40%, 15.20%, 2.40%, respectively. CYP4F2*1/*1, *1/*3, *3/*3 genotype was 50.40%, 39.20%, 10.40%, respectively. With the same genotypes for other loci, patients who carried at least one VKORC1-16398G mutant allele had increased warfarin stable daily dose compared with VKORC1-1639AA. Compared with CYP4F2*1/*1, those carrying at least one CYP4F2*3 mutant allele had warfarin stable daily dose increased by 5.9%-13.00%. The percentage of ideal prediction calculated from IWPC model (59.20%), Huang model (57.60%) and Ohno model (52.80%) were higher than others. The MAE were 0.35 (95%CI: 0.11-0.49), 0.15 (95%CI: 0.10-0.32), 0.39 (95%CI: 0.12-0.51), respectively. CONCLUSION The polymorphisms of CYP2C9, VKORC1 and CYP4F2 genes can influence the stable dose of warfarin in Shandong population. IWPC algorithm is suitable for guiding the use of warfarin in this population.
Anticoagulants ; administration & dosage ; Aryl Hydrocarbon Hydroxylases ; Cytochrome P-450 CYP2C9 ; genetics ; Cytochrome P450 Family 4 ; genetics ; Dose-Response Relationship, Drug ; Genotype ; Humans ; Models, Theoretical ; Polymorphism, Genetic ; Vitamin K Epoxide Reductases ; genetics ; Warfarin ; administration & dosage

Objective    To explore the clinical pattern of intrapulmonary lymph node metastasis and the significance of No.13 and No.14 lymph nodes biopsy in patients with non-small cell lung cancer (NSCLC). Methods    The clinical data of 234 patients with primary peripheral NSCLC who underwent systemic dissection of intrathoracic lymph nodes and intrapulmonary lymph nodes in the First Affiliated Hospital of Chongqing Medical University between 2013 and 2015 were retrospectively analyzed. There were 159 males and 75 females, aged 36-89 (61.35±8.57) years. Statistical analysis was performed accordingly on hilar (No.10), interlobar (No.11), lobar (No.12) and segmental (No.13 and 14) sites of the samples of N1 lymph nodes after surgery. Results    A total of 3 019 lymph nodes of No.10-14 were dissected in 234 patients (12.9 per patient). The 263 lymph nodes were positive with a rate of 8.71% (263/3 019) and lymph node metastasisa occured in 99 patients with a rate of 42.31% (99/234), among whom there were 40 patients of N1 metastasis, 48 of N1+N2 metastasis and 11 of N2 skipping metastasis. Routine pathological examination demonstrated No.13 and No.14 lymph nodes metastasis in 16 patients with a rate of 6.84% (16/234). In 886 dissected lymph nodes of No.13 and No.14, 86 lymph nodes showed metastasis with a rate of 9.71% (86/886). Of the patients with swelling hilar and mediastinal lymph nodes reported by preoperative CT scan, only 56.32% of them were confirmed with lymph node metastasis by postoperative histopathology; while 34.01% of the patients with normal size lymph nodes had lymph node metastasis. Conclusion    In the surgical treatment of NSCLC, it is necessary to detect the metastasis of No. 13 and 14 lymph nodes and non-tumor parabronchial lymph nodes, which is helpful to obtain accurate postoperative TNM staging and is of great significance for guiding postoperative treatment. Preoperative CT is not a reliable method to judge lymph node metastasis, particularly for intrapulmonary lymph node metastasis.

Objective To evaluate the efficacy and safety of a 755 nm picosecond Alexandrite la-ser with a diffractive lens array in the treatment of facial photoaged skin .Methods Twenty-six pa-tients with facial photoaging were recruited and received 3 treatments at 4-week intervals .Laser energy was applied over the entire face at a fixed spot size of 6 mm ,with a fluence of 0 .71 J/cm2 and 5Hz . Blinded clinical assessment was performed by 2 independent dermatologists on a 5-point global pho-toaging scale (GPS) .Patients were also questioned on the extent of improvement of rhytides ,skin tightening ,and complexion with a 4-point global aesthetic improvement scale (GAIS) and satisfaction . Adverse events were also evaluated .Results Twenty-six patients completed the treatment .Compared with the baseline ,there was a significant improvement in facial photoaged skin after 3 treatments ,and these positive outcomes were maintained up to the 3-month follow-up ,according to the GPS and GAIS scores .Moderate pain and transient erythema were observed as the two main discomforts associated with the treatment .Most patients were satisfied with the treatment .Conclusions This 755 nm pico-second Alexandrite laser with a diffractive lens array optic is effective in the treatment offacial pho -toaged skin ,and the therapy also seems safe and well tolerated .

Objective To estimate the application value of colony PCR in the detection of pathogenic filamentous fungi. Methods Colony PCR was established and performed to amplify the internal transcribed spacer (ITS) region of 19 species (strains) of filamentous fungus followed by sequencing analysis. At the same time, DNA extracts from 8 of the 19 species of filamentous fungus were subjected to conventional PCR. Hha I and Hinf I endonucleases were used for restriction fragment length polymorphism (RFLP) analysis of the conventional and colony PCR products. Comparison analysis was carried out between the colony and conventional PCR. Results Of the 19 strains, 16(84.2%) yielded positive results by colony PCR; sequence analysis of the PCR products of ITS region revealed a 96% - 100% similarity with the reference sequence (NCBI database)of corresponding fungi. The amplification product length and RFLP profile of these products from the 8 species of filamentous fungus, except for those from Aspergillus nidulans, were consistent between the colony and conventional PCR. Conclusions Compared with conventional PCR, colony PCR-based detection of filamentous fungi is easy to operate, time and labor-saving, with high accuracy and reliability, and can be applied to the rapid identification of filamentous fungi.

Objective To develop a PCR-RFLP method to rapidly identify filamentous fungi causing deep infection. Methods Universal fungal primers were used to amplify the internal transcribed spacer (ITS) region of Aspergillus fumigatus, Aspergillus Bavus, Aspergillus terreus, Aspergillus niger, Aspergillus versicolor, Aspergillus nidulans, Scedosporium apiospermum and Fusarium moniliforme followed by restriction fragment length polymorphism (RFLP) analysis with restrictive endonucleases Hha I, Hae III, Hinf I, Taq I and Msp I. Then, 22 clinical and 2 environmental fungal isolates were identified with the developed PCR-RFLP method. Results The RFLP analysis of PCR products with restrictive endonucleases Hha I and Hinf I allowed discrimination of 8 filamentous fungi causing invasive infection, and it took only 1 day to carry out the whole procedure from DNA extraction to PCR and restriction digestion. The identification results of 22 clinical strains and 2 environmental isolates with this PCR-RFLP method were completely consistent with those with conventional morphological method. Conclusion PCR-RFLP analysis is an efficient method for rapid identification of cultured filamentous fungi.