ObjectiveTo explore the effects of Renshentang on atherosclerosis （AS） in mice based on the role of transient receptor potential vanilloid1 （TRPV1） in regulating cholesterol metabolism in foam cells. MethodsNine SPF-grade 8-week-old C57BL/6J mice were set as a normal group， and 60 ApoE^-/- mice were randomized into model， positive drug （simvastatin， 0.02 g·kg^-1·d^-1）， and low-， medium-， and high-dose （1.77， 3.54， 7.08 g·kg^-1·d^-1， respectively） Renshentang groups （n=12） according to body weight. The normal group was fed with a normal diet， and the other groups were fed with a high-fat diet and given corresponding drugs by oral gavage for the modeling of AS. The mice were administrated with corresponding drugs once a day for 12 weeks. After the last administration and fasting for 12 h， the aorta was collected. Plaque conditions， pathological changes， levels of total cholesterol （TC）， triglcerides （TG）， low-density lipoprotein-cholesterol （LDL-C）， and high-density lipoprotein-cholesterol （HDL-C）， and the expression of TRPV1， liver X receptor （LXR）， inducible degrader of the low-density lipoprotein receptor （IDOL）， and low-density lipoprotein receptor （LDLR） in the aortic tissue were observed and detected by gross oil red O staining， HE staining， Western blot， immunohistochemistry， and real-time PCR. ResultsCompared with the normal group， the model group presented obvious plaque deposition in the aorta， raised levels of TC， TG， and LDL-C in the serum （P<0.01）， up-regulated expression level of LDLR in the aorta （P<0.01）， lowered level of HDL-C in the serum， and down-regulated expression levels of TRPV1， LXR， and IDOL in the aorta （P<0.05， P<0.01）. Compared with the model group， the positive drug and Renshentang at different doses alleviated AS， elevated the levels of HDL-C， TRPV1， LXR， and IDOL （P<0.05， P<0.01）， while lowering the levels of TC， TG， LDL-C， and LDLR （P<0.05， P<0.01）. ConclusionRenshentang has a lipid-lowering effect on AS mice. It can effectively reduce lipid deposition， lipid levels， and plaque area of AS mice by activating TRPV1 expression and regulating the LXR/IDOL/LDLR pathway.

ObjectiveTo explore the effects of Renshentang on atherosclerosis （AS） in mice based on the role of transient receptor potential vanilloid1 （TRPV1） in regulating cholesterol metabolism in foam cells. MethodsNine SPF-grade 8-week-old C57BL/6J mice were set as a normal group， and 60 ApoE^-/- mice were randomized into model， positive drug （simvastatin， 0.02 g·kg^-1·d^-1）， and low-， medium-， and high-dose （1.77， 3.54， 7.08 g·kg^-1·d^-1， respectively） Renshentang groups （n=12） according to body weight. The normal group was fed with a normal diet， and the other groups were fed with a high-fat diet and given corresponding drugs by oral gavage for the modeling of AS. The mice were administrated with corresponding drugs once a day for 12 weeks. After the last administration and fasting for 12 h， the aorta was collected. Plaque conditions， pathological changes， levels of total cholesterol （TC）， triglcerides （TG）， low-density lipoprotein-cholesterol （LDL-C）， and high-density lipoprotein-cholesterol （HDL-C）， and the expression of TRPV1， liver X receptor （LXR）， inducible degrader of the low-density lipoprotein receptor （IDOL）， and low-density lipoprotein receptor （LDLR） in the aortic tissue were observed and detected by gross oil red O staining， HE staining， Western blot， immunohistochemistry， and real-time PCR. ResultsCompared with the normal group， the model group presented obvious plaque deposition in the aorta， raised levels of TC， TG， and LDL-C in the serum （P<0.01）， up-regulated expression level of LDLR in the aorta （P<0.01）， lowered level of HDL-C in the serum， and down-regulated expression levels of TRPV1， LXR， and IDOL in the aorta （P<0.05， P<0.01）. Compared with the model group， the positive drug and Renshentang at different doses alleviated AS， elevated the levels of HDL-C， TRPV1， LXR， and IDOL （P<0.05， P<0.01）， while lowering the levels of TC， TG， LDL-C， and LDLR （P<0.05， P<0.01）. ConclusionRenshentang has a lipid-lowering effect on AS mice. It can effectively reduce lipid deposition， lipid levels， and plaque area of AS mice by activating TRPV1 expression and regulating the LXR/IDOL/LDLR pathway.

ObjectiveTo investigate the mechanism and effect of Qingfei Paidu decoction on transient receptor potential vanilloid-1/Transient receptor potential ankyrin1 （TRPV1/TRPA1） based on heat-sensitive channel and inflammatory response. MethodAccording to body weight， 80 8-week-old C57BL/6 mice were randomly divided into the normal group， model group， dexamethasone group （5 mg·kg^-1）， and low-dose， medium-dose， and high-dose groups of Qingfei Paidu decoction （14.865， 29.73， 59.46 g·kg^-1）， with 12 mice in each group. In addition to the normal group， the other groups were administered 20 μL （1×10^-3 g·kg^-1） to each mouse by airway infusion to establish the acute lung injury （ALI） model. In the administration group， the drug was given 1 h after modeling and again after an interval of 24 h. The lung tissue was taken 36 h after modeling. Double lung wet/dry weight ratio（W/D）， hematoxylin-eosin （HE） staining， enzyme-linked immunosorbent assay （ELISA）， and Western blot were used to observe and detect the pathological changes of lung tissue， expression levels of inflammatory cytokine tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6）， and expressions of TRPV1 and TRPA1 proteins in heat-sensitive channel， nuclear factor kappa-B （NF-κB）， inhibitor of NF-κB （IκBα） in inflammatory pathway， and phosphorylated proteins. The phosphorylated protein/total protein ratio was calculated. ResultCompared with that in the normal group， the lung tissue of mice in the model group was seriously damaged， and pulmonary capillary permeability increased. Alveolar capillary congestion and dilation destroyed the complete structure of the alveolar， and the alveolar wall thickened. A large number of inflammatory cells and red blood cells were infiltrated， and pulmonary edema was significantly aggravated. The expressions of TNF-α， IL-6， TRPV1， TRPA1， phosphorylated NF-κB p65/NF-κB p65， and phosphorylated IκBα/IκBα were significantly increased （P<0.01）， and the whole lung W/D was significantly increased （P<0.01）. Compared with the model group， the dexamethasone group and low-dose， medium-dose， and high-dose groups of Qingfei Paidu decoction could significantly improve pulmonary edema. TNF-α， IL-6， TRPV1， TRPA1， lung tissue NF-κB p65， and IκBα phosphorylated protein/total protein ratio decreased significantly （P<0.05， P<0.01）. The whole lung W/D also decreased significantly （P<0.05， P<0.01）. ConclusionQingfei Paidu decoction has anti-inflammatory and protective effects on LPS-ALI mice， which can effectively reduce inflammation， induce diuresis， and alleviate edema. Its mechanism may be related to the regulation of the expression of TRPA1 and TRPV1 and the inhibition of the activation of the NF-κB pathway.

Objective:To investigate the clinicopathological, immunohistochemical and molecular characteristics of cutaneous ALK-rearranged Spitz melanocytoma.Methods:Two cases of cutaneous ALK-rearranged Spitz melanocytoma from outside hospital consultations in Department of Pathology, Affiliated Cancer Hospital of Fudan University in August 2020 and in Shanghai Ackermann Medical Laboratory in June 2022 were collected. The clinicopathological features, immunophenotypes and molecular profiles of two patients with cutaneous Spitzoid melanocytic tumor harboring ALK-rearrangement were analyzed. The literatures were reviewed.Results:The study included an 8-year-old boy and an 11-year-old girl, who presented with a polypoid lesion in the skin of right thigh and left auricle measuring 1.0 cm and 1.2 cm, respectively. Histologically, they were composed of medium to large-sized epithelioid to plump spindle cells, arranged in nested, plexiform or fascicular patterns in the superficial dermis. The neoplastic cells had abundant eosinophilic cytoplasm with round to ovoid vesicular nuclei containing prominent eosinophilic nucleoli. One case showed mild to moderate nuclear pleomorphism and mitotic activity (average, 2/mm 2). Immunohistochemically, the epithelioid and plump spindle cells showed diffuse and strong staining of S-100 protein, SOX10, and ALK (D5F3 and 1A4), but did not express HMB45, PNL2 and MiTF. ALK-rearrangement was detected by fuorescence in situ hybridization in both cases. Subsequent next generation sequence (NGS) analysis identified KANK1::ALK and TPM3:ALK fusions. At 34 and 14 months after surgical resection, both patients remained well with no signs of recurrence or metastasis. Conclusions:ALK-rearranged Spitz melanocytoma represents a morphologically and genetically distinct subset of Spitz melanocytoma, characterized clinically by predilection in children and adolescents, with Spitzoid morphology in plexiform pattern, positive immunohistochemical stains, and rearrangement of ALK. As some cases show atypical features and high mitotic activity, a distinction from Spitz melanoma is warranted.

Epigenomic imbalance drives abnormal transcriptional processes,promoting the onset and progression of cancer.Although defective gene regulation generally affects carcinogenesis and tumor suppression networks,tumor immunogenicity and immune cells involved in antitumor responses may also be affected by epigenomic changes,which may have significant implications for the development and application of epigenetic therapy,cancer immunotherapy,and their combinations.Herein,we focus on the impact of epigenetic regulation on tumor immune cell function and the role of key abnormal epigenetic processes,DNA methylation,histone post-translational modification,and chromatin structure in tumor immunogenicity,and introduce these epigenetic research methods.We emphasize the value of small-molecule inhibitors of epigenetic modulators in enhancing antitumor immune responses and discuss the challenges of developing treatment plans that combine epigenetic therapy and immuno-therapy through the complex interaction between cancer epigenetics and cancer immunology.

The genomic DNA of Rubus rosaefolius was extracted and sequenced by Illumina NovaSeq platform to obtain the complete chloroplast genome sequence, and the sequence characteristics and phylogenetic analysis of chloroplast genes were carried out. The results showed that the complete chloroplast genome of the R. rosaefolius was 155 650 bp in length and had a typical tetrad structure, including two reverse repeats (25 748 bp each), a large copy region (85 443 bp) and a small copy region (18 711 bp). A total of 131 genes were identified in the whole genome of R. rosaefolius chloroplast, including 86 protein coding genes, 37 tRNA genes and 8 rRNA genes. The GC content of the whole genome was 36.9%. The genome of R. rosaefolius chloroplast contains 47 scattered repeats and 72 simple sequence repeating (SSR) loci. The codon preference is leucine codon, and the codon at the end of A/U is preferred. Phylogenetic analysis showed that R. rosaefolius had the closest relationship with R. taiwanicola, followed by R. rubraangustifolius and R. glandulosopunctatus. The chloroplast genome characteristics and phylogenetic analysis of R. rosaefolius provide a theoretical basis for its genetic diversity research and chloroplast development and utilization.
Phylogeny ; Rubus/genetics* ; Genome, Chloroplast ; Fruit/genetics* ; Codon/genetics*

Objective:To investigate the feasibility of construction of rat models of psoriasis-like lesions by using interleukin (IL) -23/T-helper 17 (Th17) axis-related cytokines combined with imiquimod.Methods:A total of 110 Wistar rats were randomly divided into normal control group, imiquimod alone group, imiquimod combined with interferon (IFN) -α2a (180 000, 60 000, 20 000 IU/kg) groups, imiquimod combined with tumor necrosis factor (TNF) -α (45 000, 15 000, 5 000 IU/kg) groups, and imiquimod combined with IL-2 (90 000, 30 000, 10 000 IU/kg) groups, and there were 10 rats in each group. After hair removal from the central area (2 cm × 2 cm) of the rat back, rats in the imiquimod alone group were topically treated with imiquimod 5% cream at a dose of 20 mg/cm 2 on the shaved back; rats in the imiquimod combined with different cytokine groups were treated with topical imiquimod 5% cream at the same dose on the shaved back for 15 minutes followed by intraperitoneal injections of cytokines at corresponding doses once a day for 10 consecutive days. During the treatment, skin lesions on the rat back were evaluated by using the psoriasis area and severity index (PASI) scores every day. On day 10, serum samples were collected from the rats after anesthesia, and enzyme-linked immunosorbent assay (ELISA) was performed to detect levels of IL-17A, TNF-α, IL-23, IFN-α and IL-1β in the serum samples in each group; then, the rats were sacrificed, lesional skin tissues on the rat back were taken for histopathological examinations and evaluated by Baker scores; an immunohistochemical study was conducted to determine the expression of CD4 and CD8 in some skin lesions. One-way analysis of variance was used for comparisons among multiple groups, and least significant difference (LSD) - t test for multiple comparisons; for data with heterogeneous variance, the Kruskal-Wallis H test was used. Results:On day 3 after molding, the rats in the imiquimod alone group and combination groups gradually presented with psoriasis-like skin manifestations, such as erythema, scales and epidermal thickening; the PASI scores reached a peak on day 7 in the imiquimod alone group, and on day 6 in the combination groups. On day 10, histopathological examination of the skin lesions in the imiquimod alone group and combination groups both showed different psoriasis-like pathological features, such as hyperkeratosis, parakeratosis, acanthosis, thinning or disappearance of the granular layer. There were significant differences in the PASI scores and Baker scores among the normal control group, imiquimod alone group and combination groups ( H = 43.33, F = 42.15, both P < 0.001). The PASI scores were higher in the imiquimod combined with IFN-α2a (180 000 IU/kg) group and the imiquimod combined with IL-2 (90 000 IU/kg) group (9.4 ± 1.1, 8.8 ± 0.6, respectively) than in the imiquimod alone group (7.5 ± 1.1, P = 0.002, 0.030 respectively) ; the Baker scores were higher in the imiquimod combined with IFN-α2a (180 000, 60 000 IU/kg) groups, the imiquimod combined with TNF-α (45 000 IU/kg) group, and the imiquimod combined with IL-2 (90 000 IU/kg) group than in the imiquimod alone group (all P < 0.05). The serum levels of TNF-α, IL-17A, IL-1 β and IL-23 significantly differed among groups ( F = 128.97, F = 6.90, H = 27.45, H = 21.10, all P < 0.05). Compared with the imiquimod alone group, the IL-17A level significantly increased in the imiquimod combined with IL-2 (30 000 IU/kg) group (5.54 ± 1.78 pg/ml vs. 4.20 ± 1.14 pg/ml, P = 0.009), and the IL-23 level significantly increased in the imiquimod combined with IL-2 (90 000 IU/kg) group (37.89 ± 32.85 pg/ml vs. 8.56 ± 6.08 pg/ml, P = 0.036). Immunohistochemical study showed significant differences in the expression of CD4 and CD8 in skin lesions among all groups ( F = 7.21, H = 18.32, both P < 0.001), and the expression of CD4 and CD8 in skin lesions was significantly higher in the imiquimod combined with IL-2 (90 000 IU/kg) group than in the imiquimod alone group ( t = -2.46, -2.32, respectively, both P < 0.05) . Conclusion:Imiquimod combined with IFN-α2a or IL-2 could promote the occurrence of psoriasis-like skin lesions in rats, aggravate the development of psoriasis and prolong the maintenance time of the rat models.

Objective:To detect the expression of has_circ_0001459 in peripheral blood mononuclear cells (PBMCs) of Epstein-Barr virus (EBV)-infected individuals and evaluate its diagnostic value for EBV infection.Methods:The expression profiles of circRNAs in PBMCs of patients with EBV infection were obtained by whole-transcriptome sequencing. Differentially expressed circRNAs with statistical significance were selected based on the criteria of ∣log 2Fold Change∣≥1 and P<0.05. Based on the composition, length and primer specificity, hsa_circ_0001459 with high expression was selected for further research. The relative expression of hsa_circ_0001459 in PBMCs of 60 patients with EBV infection and 45 healthy people was detected by real-time fluorescent quantitative PCR. Receiver operating characteristic (ROC) curve and Kappa analysis were used to analyze the diagnostic value of hsa_circ_0001459 expression for EBV infection. Results:The expression of hsa_circ_0001459 showed an up-regulation trend in patients with EBV infection as compared with that in the healthy people, and the results were consistent with the sequencing results. The area under the ROC curve for screening EBV-infected individuals was 0.83(95%CI: 0.75-0.91) with the sensitivity of 0.80(95%CI: 0.66-0.89) and the specificity of 0.77(95%CI: 0.65-0.86). Kappa analysis indicated that hsa_circ_0001459 was moderately consistent with EBV DNA in the diagnosis of EBV infection (κ=0.51).Conclusions:Hsa_circ_0001459 tended to be highly expressed in PBMCs of EBV-infected individuals, which might be uses as a diagnostic marker for EBV infection.

Objective:To investigate the clinical value of shear wave elastography (SWE) in predicting pathological responses to neoadjuvant chemotherapy in breast cancer.Methods:According to the postoperative pathological responses, 56 patients who received neoadjuvant chemotherapy followed by surgical excision in the Fujian Cancer Hospital from August 2019 to September 2020 were divided into responders and non-responders. The relative change rates of tumor maximum diameter(ΔD2, ΔD4) and SWE stiffness (ΔEmax2, ΔEmax4, ΔEmean2, ΔEmean4) were assessed before NAC and after different NAC cycles (t2, t4). Clinical information, including age, T, N stages, ER, PR, HER2, Ki67, and molecular subtype were also considered as the variables. The independent influencing factors of pathological responses after neoadjuvant chemotherapy were obtained by logistic regression analysis and diagnostic test was carried out.Results:There were 23 cases as responders (41.0%, 23/56), and 33 cases as non-responders (58.9%, 33/56). Results of multivariate analysis showed ΔEmax4 and HER2 index were independent influencing factors of pathological responses ( OR=1.11, P<0.001; OR=31.81, P=0.002). Area under curve of the ΔEmax4 (AUC: 0.869, 95% CI: 0.746-0.941) was higher than that of HER2 (AUC: 0.690, 95% CI: 0.545-0.834). The combination of ΔEmax4 and HER2 gave the best prediction of pathological responses (AUC 0.930, 95% CI: 0.829-0.981). the sensitivity, specificity, diagnostic accuracy, postive predictive value, and negative predictive value were 78.26%, 96.97%, 75.23%, 94.73%, and 86.49%, respectively. Conclusions:ΔEmax4 and HER2 are independent predictors of pathological responses after neoadjuvant chemotherapy for breast cancer. Combined ΔEmax4 and HER2 can improve the predictive diagnostic efficacy of pathological responses to chemotherapy for breast cancer.

Objective:To compare the detection rate of genital Chlamydia trachomatis (CT) DNA between urine and urethral/cervical swab samples. Methods:From December 2018 to December 2019, a total of 1 475 outpatients were collected from sexually transmitted disease clinics in 7 medical institutions, such as Department of Venereology, Guangzhou Institute of Dermatology, including 1 118 males and 357 females. One urethral/cervical swab sample and one urine sample were collected successively from each patient. Real-time fluorescence-based PCR was performed to detect CT DNA in urine and urethral/cervical swab samples, and paired chi-square test was used to compare the positive rate of CT DNA between the 2 kinds of samples. Random- or fixed-effect meta-analysis was conducted for the test of heterogeneity and merging of positive rates of CT DNA in the urine and urethral/cervical swabs among 7 medical institutions.Results:The positive rate of CT DNA in the urine samples was significantly higher than that in the swab samples from 4 medical institutions (all P < 0.05) , while there was no significant difference in the positive rate of CT DNA between the 2 kinds of samples from 3 medical institutions (all P > 0.05) . The heterogeneity ( I2) estimates of the CT-DNA positive rate in urine and swab samples among different medical institutions were 78.6% (95% CI: 55.9% - 89.6%) and 73.7% (95% CI: 43.7% - 87.7%) , respectively; meta-analysis showed that the total merged positive rate of CT DNA in the urine samples was 10.8% (95% CI: 7.2% - 15.9%) , which was significantly higher than that in the swab samples (7.8%, 95% CI: 4.9% - 12.1%; χ2 = 39.2, P < 0.05) . Compared with the swab sample-based CT-DNA detection method, the sensitivity, specificity, positive predictive value, negative predictive value and consistency rate of the urine sample-based CT-DNA detection method were 97.0% (128/132) , 96.3% (1 293/1 343) , 71.9% (128/178) , 99.7% (1 293/1 297) , and 96.3% (1 421/1 475) , respectively. The positive rate of CT DNA in the urine samples from 1 118 male patients was 11.0% (95% CI: 7.2% - 16.5%) , which was significantly higher than that in the swab samples (7.6%, 95% CI: 4.9% - 11.8%; χ2 = 34.3, P < 0.05) . There was no significant difference in the positive rate of CT DNA between the urine (11.9%, 95% CI: 7.7% - 17.9%) and cervical swab samples from 357 female patients (10.4%, 95% CI: 7.6% - 14.0%; χ2 = 3.2, P > 0.05) . Conclusions:The positive rate of CT DNA in urine samples is higher than or similar to that in urethral/cervical swab samples. The urine sample-based CT-DNA detection method has characteristics of convenience, non-invasiveness, painlessness and low cost, and is worthy of clinical promotion.