circPTK2 promotes proliferation, migration and invasion of trophoblast cells through the miR-619/WNT7B pathway in preeclampsia.

It has been shown that the circular RNA (circRNA) circPTK2 modulates many types of diseases. However, the possible functions as well as the molecular mechanisms of circPTK2 in preeclampsia (PE) and their effects on trophoblast are unknown. Herein, we obtained the placental tissues from 20 pregnant women with PE who delivered in the Yueyang Maternal Child Medicine Health Hospital between 2019 and 2021 to serve as the PE group, and a normal group was composed of 20 healthy pregnant women with normal prenatal examinations. The circPTK2 level was significantly reduced in tissues from the PE group. The expression and localization of circPTK2 were verified using RT-qPCR. CircPTK2 silencing inhibited HTR-8/SVneo growth and migration in vitro. To investigate the underlying mechanism of circPTK2 in PE progression, dual-luciferase reporter assays were conducted. It was found that circPTK2 and WNT7B could bind directly to miR-619, and that circPTK2 affected WNT7B expression by sponging miR-619. To conclude, this study identified the functions and mechanisms of the circPTK2/miR-619/WNT7B axis in PE progression. In this way, circPTK2 has the potential to be used both in diagnostic and therapeutic settings for PE.

PCDH9 suppresses melanoma proliferation and cell migration.

Background: Melanoma has dramatically increased during last 30 years with low 5-year survival and prognosis rate. Methods: Melanoma cells (A375 and G361) were chosen as the in vitro model. The immunohistochemical (IHC) analysis and bioinformatics mining exhibited the suppression of PCDH9 on melanoma. The interference and overexpression of PCDH9 were infected by lentivirus. The effects of PCDH9 on melanoma cells were assessed in terms of alteration of PCDH9 such as cell viability, apoptosis, cell cycle, and wound-healing assay. Moreover, expressions of PCDH9 with other genes (MMP2, MMP9, CCND1, and RAC1) were also assessed by PCR. Results: The alteration of PCDH9 has a negative correlation with MMP2, MMP9, and RAC1 but had a positive correlation with CCND1 (Cyclin D1) and apoptosis. Increase of PCDH9 could suppress melanoma cells and inhibit migration but not exert significant effects on cell cycle. IHC showed lower PCDH9 expression in melanoma tissue with main expression in cytoplasm. Conclusion: Overexpressed PCDH9 suppressed melanoma cells, and PCDH9 can be considered as an independent prognostic factor for melanoma; even re-expression of PCDH9 can serve as a potential therapeutic strategy for melanoma treatment.

Circ_0084043-miR-134-5p axis regulates PCDH9 to suppress melanoma.

The low survival rates, poor responses, and drug resistance of patients with melanoma make it urgent to find new therapeutic targets. This study investigated whether the circ_0084043-miR-134-5p axis regulates the antitumor effect of protocadherin 9 (PCDH9) in melanoma. Ectopic expression or knock down (KD) of PCDH9 with a lentivirus vector, we explored its effects on the proliferation, invasion, and apoptosis of melanoma and verified its regulatory effect on ras-related C3 botulinum toxin substrate 1 (RAC1), proline-rich tyrosine kinase 2 (Pyk2), Cyclin D1, matrix metalloproteinase 2 (MMP2), and MMP9. We further observed the effect of KD circ_0084043 on the malignant behavior of melanoma and studied whether circ_0084043 sponged miR-134-5p and regulated PCDH9. We found that circ_0084043 was overexpressed in melanoma and associated with the malignant phenotype. PCDH9 was poorly expressed in human melanoma tissues, and overexpression of PCDH9 inhibited melanoma progression. Quantitative real-time PCR and Western blotting results showed that overexpression of PCDH9 could downregulate RAC1, MMP2, and MMP9 and upregulate Pyk2 and Cyclin D1. Circ_0084043 KD inhibited invasion and promoted apoptosis in melanoma cells. Circ_0084043 could sponge miR-134-5p and thus indirectly regulate PCDH9. Furthermore, we discovered that inhibiting circ_0084043 had an anti-PD-Ll effect. In vivo, PCDH9 overexpression inhibited melanoma tumor growth, but PCDH9 KD promoted it. In conclusion, PCDH9, which is regulated by the circ 0084043-miR-134-5p axis, can suppress malignant biological behavior in melanoma and influence the expression levels of Pyk2, RAC1, Cyclin D1, MMP2, and MMP9.

The role of estrogen in circular RNA and metabonomics in a Neisseria gonorrhoeae infection model.

Background: Previous study shows that estrogen exerts both immunosuppressive and immunostimulative effects. Methods: In this study, estrogen was added to a Neisseria gonorrhoeae infection model, and transcriptome sequencing and metabolomics studies were performed to clarify the changes in circular RNA (circRNA) and metabolic pathways regulated by the addition of estrogen. Results: The results showed that following the addition of estrogen to the gonococcal infection model, the expression of circRNAs was up-regulated and the expression of circRNAs was down-regulated. In the metabolic group, it was found that after the addition of estrogen, the expression of nine metabolites was down-regulated and 61 metabolites were up-regulated. Furthermore, through network interaction analysis of differentially-expressed circRNAs and differentially-expressed metabolites, we found that the top 10 significantly related metabolites and circRNA were 2-Epoxybutane/novel_circ_0024520; 1,2-Epoxybutane/novel_circ_0061793; 2-Imino-4-methylpiperidine/novel_circ_0012178; 2-Imino-4-methylpiperidine/novel_circ_0056959; Acetone oxime/novel_circ_0012178; Adifoline/novel_circ_0012178; CARBETAPENTANE/novel_circ_0054387; CARBETAPENTANE/novel_circ_0056959; deoxy-PF1140/mmu_circ_0000397; and Methyl (2E,6Z)-dodecadienoate/novel_circ_0012178. Among these, CARBETAPENTANE/novel_circ_0054387 and CARBETAPENTANE/novel_circ_0056959 were positively correlated, while the remaining metabolites were negatively correlated. Conclusions: In this study, high-throughput sequencing and metabolomics mass spectrum were applied to screen the differentially-expressed circRNAs and metabolites regulated by estrogen, which will help to provide new research ideas and indicators for asymptomatic infections in women, and can be meaningful for the relevant study in the future.

Widely targeted metabolomics reveals stamen petaloid tissue of Paeonia lactiflora Pall. being a potential pharmacological resource.

Paeonia lactiflflora Pall. has a long edible and medicinal history because of the very high content of biologically active compounds. However, little information is available about the metabolic basis of pharmacological values of P. lactiflora flowers. In this study, we investigated metabolites in the different parts of P. lactiflora flowers, including petal, stamen petaloid tissue and stamen, by widely targeted metabolomics approach. A total of 1102 metabolites were identified, among which 313 and 410 metabolites showed differential accumulation in comparison groups of petal vs. stamen petaloid tissue and stamen vs. stamen petaloid tissue. Differential accumulated metabolites analysis and KEGG pathway analysis showed that the flavonoids were the most critical differential metabolites. Furthermore, difference accumulation of flavonoids, phenolic acids, tannins and alkaloids might lead to the differences in antioxidant activities and tyrosinase inhibition effects. Indeed, stamen petaloid tissue displayed better antioxidant and anti-melanin production activities than petal and stamen through experimental verification. These results not only expand our understanding of metabolites in P. lactiflora flowers, but also reveal that the stamen petaloid tissues of P. lactiflora hold the great potential as promising ingredients for pharmaceuticals, functional foods and skincare products.

[Clinical Study on Dynamic Detection of Minimal Residual Disease in Acute Myeloid Leukemia by Multiparameter Flow Cytometry].

OBJECTIVE: To investigate the prognostic significance of dynamic detection of minimal residual disease (MRD) in patients with acute myeloid leukemia (AML) by 8-color flow cytometry. METHODS: MRD of 282 AML patients who achieved remission after initial therapy was detected by 8-color flow cytometry. MRD threshold for predicting recurrence was determined by receiver operating characteristic (ROC) curve, and time from MRD-positive to clinical recurrence was analyzed. The differences in overall survival (OS) time and relapse-free survival (RFS) time of patients with different MRD-changes were compared, and the related factors of recurrence in patients with MRD-negative were analyzed by univariate and logistic regression analysis. RESULTS: ROC curve determined that the MFC-MRD threshold for predicting the recurrence of AML was 0.105%, and the recurrence rate of MRD-positive patients was significantly higher than that of MRD-negative patients [52.45% (75/143 cases) vs 35.97% (50/139 cases), P=0.005]. The patients in MRD persistent positive group and negative to positive group recurred earlier than those in positive to negative group and negative-positive fluctuation group (P<0.005). Survival analysis showed that OS and RFS time of patients with MRD persistent positive were significantly shorter than those of patients with MRD persistent negative, positive to negative, and negative-positive fluctuation (P<0.005). There was no significant difference in OS and RFS between MRD negative to positive group and MRD persistent positive group (P>0.005), either between MRD persistent negative group and MRD positive to negative group (P>0.005). Among 139 MRD-negative patients, 50 recurred. Univariate and logistic regression analysis showed that the risk of recurrence increased with the increase of white blood cells level (95%CI: 1.000-1.013, P=0.045). The risk of recurrence in patients without hematopoietic stem cell transplantation (HSCT) was 9.694 times higher than that in patients who received HSCT (95%CI: 1.720-54.651, P=0.010), and in the high-risk group was 5.848 times higher than that in the low-risk group (95%CI: 1.418-24.121, P=0.015). CONCLUSION: The prognosis of AML patients with different MRD changes is significantly different. No matter MRD-positive or MRD-negative at the initial remission, dynamic detection of MRD after treatment is more helpful to accurately guide treatment.

A trinity fingerprint evaluation system of traditional Chinese medicine.

This study focused on developing a set of quality evaluation methods that can reflect the multi-levels and multi-characteristics of traditional Chinese medicine (TCM). Taking licorice (Glycyrrhiza glabra L.) as the method development sample, the feasibility of multi-markers assay by monolinear method (MAML) was explored through the standard curve relationship among active components for the first time. Using glycyrrhizic acid as measurement marker, MAML can simultaneously quantify five components of licorice, including isoliquiritigenin, isoliquiritin apioside, liquiritigenin, liquiritin and liquiritin apioside. Comparing MAML and quantitative analysis of multi-components by single-marker (QAMS) to the external standard method (ESM) respectively, it was found that there was no significant difference in the content of components that were calculated by MAML and ESM (the relative error (RE) was generally less than 2.00%). However, the RE of the component content calculated by QAMS fluctuated greatly, indicating that the MAML was more accurate than QAMS. In addition, UV and THz quantum fingerprints were initiated by the interval erasure method. Taking the systematically quantified fingerprint method as the core, a "Trinity" fingerprint quality evaluation system based on HPLC, UV and THz was developed. The system successfully distinguished the quality differences of licorice samples from 13 producing areas and two ecological models by the comprehensive evaluation results. Simultaneously, the quality information of licorice at different technical levels was revealed. Finally, bivariate correlation analysis was used to examine the linkage between UV/HPLC and antioxidant spectrum efficacy, and the two-dimensional activity spectrum of licorice was provided. It may furnish a more thorough and objective analytical technique for licorice and even other TCMs in chemical fingerprint features, chemical bond vibration characteristics and biological activity information.

Evaluating the quality consistency of Keteling capsules by three-dimensional quantum fingerprints and HPLC fingerprint.

Keteling capsules (KCs), as a kind of Traditional Chinese Medicine (TCM), have been widely used in curing cough and relieving asthma. However, the complicated compositions make it challenging to evaluate their quality consistency by common methods. Herein, we explored comprehensive and efficient strategies by combining the multiple techniques to monitor and assess the characteristics of KCs. We employed the fingerprints and corresponding quantum fingerprints by fourier transform infrared spectroscopy (FT-IR), ultraviolet (UV), and differential scanning calorimetry (DSC). The antioxidant activity profiles were also studied combined with the result of three-dimensional quantum fingerprints and showed a good correlation with the internal structure and physical-chemical state. Furthermore, the 17 samples were separated and identified simultaneously by HPLC quantitative fingerprint, of which four active ingredients (chlorogenic acid, p-coumaric acid, vitexin and isovitexin) were quantitatively determined. The 17 samples were successfully classified into different grades by the systematically quantified fingerprint method (SQFM) and the quality of the samples was integrated according to the mean algorithm. The mean algorithm fusion of different evaluation techniques was compared to reveal the relationship between them, which indicated the effective improvement in accuracy and integrality. The combination of multiple analytical techniques developed in this study would effectively improve the existing single analytical methods and provide new strategy for drug quality consistency control.

Fabrication of actiniae-like atomically thin hydroxylation boron nitride@polyaniline hierarchical composites with adjustable high thermal conductivity and electrical conductivity.

Polyaniline (PANI) has been studied as soft electronic materials, which is still subject to performance obstacles such as low thermal conductivity and undesirable electrical conductivity. Herein, we report thein situpreparation of an atomically thin hydroxylated boron nitride (HO-BNNS)@PANI actiniae-like layered composite. HO-BNNS@PANI composite obtains brilliant electrical and thermal conductivity without destroying the pH sensitivity of PANI. In this case, the test results show that when the HO-BNNS content is 15 wt%, the conductivity of the HO-BNNS@PANI composite is 10.8 S cm-1, and the thermal conductivity is 1.21 W m-1·K-1(≈520% that of pure PANI). More strikingly, the HO-BNNS@PANI composite maintains the pH responsiveness of the intrinsic PANI. This greatly improves the application range of composite materials. Meanwhile, since actiniae-like structural factors simultaneously improve ion diffusion capability and optimize reaction area, after five times of doping and dedoping, the conductivity of the HO-BNNS@PANI composite can still be maintained above 60%.

Facile in situ synthesis of silver nanocomposites based on cellulosic paper for photocatalytic applications.

In this work, various photocatalysts were synthesized with an impregnation-precipitation process to in situ decorate Ag-based nanoparticles (NPs, including Ag3PO4, AgCl, Ag2O, and Ag2CO3) on the cellulosic paper. The structure and properties of the Ag-based composites were characterized by scanning electron microscopy, X-ray diffraction, transmitting electron microscopy, UV-vis diffuse reflectance spectra, and photocatalysis testing. The results showed that cellulosic paper is an efficient carrier which is feasible to grasp NPs due to the cellulosic nanofiber-network microstructure. Among the obtained samples, Ag2CO3 and AgCl NPs on cellulosic paper displayed high photocatalytic activity for the degradation of methyl orange under ultraviolet and visible light. However, photo lability of Ag2CO3 limits its recyclable. AgCl showed a better reutilization with the assistance of a surface plasmon resonance effect by Ag NPs that were grown in situ on the AgCl NPs, which formed Ag@AgCl nanocomposite structure. The photocatalytic activity of the AgCl/cellulosic paper decreased only slightly after three runs of photodegradation of methyl orange. The possible mechanism for photocatalysis was proposed. This work may provide a new method for the design of silver-based NPs/cellulosic paper nanocomposite photoreactors with favorable photocatalytic activities for industrial applications.

Primary acral amelanotic melanoma: A rare case report.

The aim of the present study was to present a rare case of primary acral amelanotic malignant melanoma (AMM). A 61-year-old man developed an aggressive tumor in the front part of the sole of his left foot, which continued to increase in size for >1 year. The biopsy results revealed epidermis loss, ulcer formation, and the presence of abundant allotropic tumor cells throughout the dermis, with deeply stained nuclei, light reddish cytoplasm and visible multinucleated giant cells with heterogeneous nuclear division. The tumor cells exhibited partial formation of nests and bundled distribution, and there were no observed pigment particles. The diagnosis was confirmed as AMM based on the findings of the histopathological examination and immunohistochemical staining for Ki67 (+++), Melan-A (+++), human melanoma black 45 (+), CD20 (-), cytokeratin (CK)7 (-) and CK5/6 (-).

Resveratrol inhibits the proliferation of melanoma cells by modulating cell cycle.

The aim of this study was to evaluate the inhibitory effects of resveratrol (RSV) in A375 and A431 melanoma cells, by assessing cell viability (CCK-8 assay), apoptosis through flow cytometer and cell morphology, cell cycle assay by flow cytometer and western blot (Cyclin D1, Rac1 and PCDH9). Our results demonstrated that RSV strongly inhibited A375 cells proliferation, by decreasing cell viability, promoting apoptosis and arresting cell cycle. Besides, to clarify the main factor - duration or concentration of RSV, the double variance analysis of independent factors was operated after Bartlett's test for homogeneity by R project. According to the outcomes obtained from statistical analyses, Cyclin D1 and PCDH9 were strongly affected by RSV duration while Rac1 was not influenced. In conclusion, RSV can inhibit A375 proliferation and trigger apoptosis by influencing cell cycle proteins; for these effects, treatment duration of RSV played more important role than concentration.

Better outcomes of modified myeloablative conditioning without antithymocyte globulin versus myeloablative conditioning in cord blood transplantation for hematological malignancies: A retrospective (development) and a prospective (validation) study.

Cord blood transplantation (CBT) is an effective option for treating hematological malignancies, but graft failure (GF) remains the primary cause of therapy failure. Thus, based on myeloablative conditioning (MAC) of busulfan with cyclophosphamide (Bu/Cy) or total body irradiation with Cy (TBI/Cy), fludarabine (Flu) was added to Bu/Cy and cytarabine (CA) to TBI/Cy for a modified myeloablative conditioning (MMAC). To compare the prognosis of MMAC with MAC, we conducted a retrospective study including 58 patients who underwent CBT with MAC or MMAC from 2000 to 2011. Neutrophil and platelet engraftment rate, overall survival (OS) and disease free survival (DFS) were significantly higher in the MMAC group (adjusted hazard ratio [HR], 2.58, 2.43, 0.36 and 0.37; p < 0.01, p = 0.01, p = 0.02 and p = 0.02, separately). Nonrelapse mortality (NRM) was comparable (p = 0.183). To validate the outcomes noted in the MMAC group, we conducted a prospective single-arm clinical trial including 188 patients who underwent CBT with MMAC from 2011 to 2015. Engraftment rate, survival and NRM of the MMAC group in the prospective trail (MMAC-P) were similar to the MMAC group in the retrospective study (MMAC-R). This study is the first to demonstrate the superiority of MMAC to MAC in CBT for hematological malignancies.

[Correlative study between JAK2 mutation and thrombosis in patients with myeloproliferative neoplasm].

OBJECTIVE: To investigate the frequency and clinical implication of JAK2 mutation in patients with myeloproliferative neoplasm(MPN)and the correlation between the mutation and thrombosis. METHODS: The clinical and laboratory data of 107 MPN patients was retrospectively analyzed. JAK2 mutation were detected with allele-specific polymerase chain reaction (AS-PCR) and sequencing. The significance of the mutation in disease diagnosis and molecular pathogenesis and the correlation between the mutation and thrombosis was analysed. RESULTS: JAK2 mutation was detected in 71 (66.4%) and thrombosis in 34 (31.8%) of the 107 MPN patients. Thrombosis occurred in 34.8% (16/46) of polycythemia vera (PV), 32.6% (14/43) of essential thrombocythemia (ET), and 22.2% (4/18) of primany myelofibrosis (PMF) patients. The difference among the 3 groups was not significant (χ(2) = 0.96, P > 0.05). The frequency of thrombosis in JAK2(+) MPN patients (82.4%, 28/34) was higher than that in JAK2(-) MPN patients (17.6%, 6/34) (χ(2) = 5.71, P < 0.05). The frequency of thrombosis in MPN patients > 60 years was higher (41.5%, 27/65) than that in patients < 60 years (16.7%, 7/42) (χ(2) = 7.28, P < 0.01). CONCLUSION: JAK2 V617F mutation occurs in significant percentage of Chinese patients with MPN. Patients with JAK2 mutation and older age are more succeptible to thrombosis. JAK2 mutation screening in patients with unknown thrombosis is helpful to reveal the underlying latent-MPN.

[Unrelated cord blood transplantation in adult patients with hematologic malignancies].

OBJECTIVE: To analyse the engraftment, transplant-related complications and survival after unrelated cord blood transplantation (UCBT) in patients with hematologic malignancies. METHODS: Twenty eight consecutive adult patients with hematological malignancies were treated with UCBT and 20 of them were advanced-stage diseases. Double or multiple UCB grafts were used for 18 patients, while single UCB graft for 10 patients. Myeloablative conditioning regimens were given to 26 cases and nonmyeloablative regimens to 2 cases. All patients were given a combination of cyclosporine (CsA) and mycophenolate mofetil (MMF) for graft-versus-host disease (GVHD) prophylaxis. RESULTS: Median time to neutrophil engraftment (≥ 0.5 × 10(9)/L) in 26 patients was 18 (14 - 37) days and platelet engraftment (≥ 20 × 10(9)/L) in 22 patients was 30 (25 - 49) days. Chimerism was weekly assessed by PCR analysis of short tandem repeat (STR) sequences in whole blood or bone marrow and 22 cases were confirmed of fully donor chimeric from 7 to 21 days after transplantation. Eighteen cases developed acute GVHD, greater than grade II in 1, and 6 of 22 patients who survived more than 100 days developed limited chronic GVHD. Eighteen cases were alive in hematologic remission at a median follow-up of 9.5 (2.5 - 72.0) months. The probability of event-free survival at 3 years was 56.7%. Two cases relapsed and 8 of 10 cases died of transplant related complications. CONCLUSIONS: UCBT could be safely and effectively used for adult patients with hematologic malignancies. Use of double UCB units is a strategy extending the feasibility of UCBT.

HLA-matched sibling transplantation with G-CSF mobilized PBSCs and BM decreases GVHD in adult patients with severe aplastic anemia.

BACKGROUND: Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is an effective treatment for severe aplastic anemia (SAA). However, graft failure and graft-versus-host disease (GVHD) are major causes of the early morbidity in Allo-HSCT. METHODS: To reduce graft failure and GVHD, we treated fifteen patients with SAA using high- dose of HSCT with both G-CSF mobilized PB and BMSCs from HLA-identical siblings to treat patients with SAA. RESULTS: All patients had successful bone marrow engraftment. Only one patient had late rejection. Median time to ANC greater than 0.5 × 10(9)/L and platelet counts greater than 20 × 10(9)/L was 12 and 16.5 days, respectively. No acute GVHD was observed. The incidence of chronic GVHD was 6.67%. The total three-year probability of disease-free survival was 79.8%. CONCLUSION: HSCT with both G-CSF mobilized PB and BMSCs is a promising approach for heavily transfused and/or allo-immunized patients with SAA.

[Intracellular cytokine expression characteristics of activated T lymphocytes in patients with acute myeloid leukemia].

T lymphocytes are an integral part of the effective immune response against various tumors, but they are frequently functionally unresponsive in tumor-bearing patients. This study was aimed to investigate the T lymphocyte function of patients with acute myeloid leukemia (AML) in different status through analyzing intracellular cytokine characteristics of T lymphocytes in AML patients. The T lymphocytes from 18 de nuevo AML patients in different status and 10 healthy controls were stimulated with phorbol 12-myristate 13-acetate (PMA) and ionomycin in the presence of monensin, and were stained with fluorescent McAbs CD4-PITC, CD8-FITC and IFNgamma-PE, ILA-PE, then their T lymphocytes were analyzed by flow cytometry. The results showed that IFN-gamma level in CD4+ and CD8+ T cells of de novo AML patients was significantly lower as compared with healthy controls (p<0.05), while IL-4 level in CD4+ and CD8+ T cells was low too, there was no significant difference between de novo AML patients and healthy controls. In AML patients in clinical remission, IFNgamma level in CD8+ T cells was significantly higher than that in de novo AML patients (p<0.05), but there was no significant difference as compared with healthy controls (p>0.05). In relapsed AML patients, IFNgamma level in CD4+ and CD8+ T cells was significantly lower than that in healthy controls and in AML patients with CR (p<0.05), while IL-4 level was significantly higher than that in healthy controls and de nuevo AML patients (p<0.05). It is concluded that the cytokine secretion in T cell subsets of AML patients in different status is changed. Correspondingly, the Th1/Tc1 level is low after stimulating CD4+ and CD8+ T cells in peripheral blood of de novo AML patients, but not different from healthy controls. Though the Th1 level in T cells of AML patients in complete remission is low, but Tc1 response is even enhanced as high as the healthy controls. The Th2/Tc2-like response of CD4+ and CD8+ T cells in relapsed AML patients obviously increases when compared with Th1/Tc1-like response.