Synthesis of Heterocyclic Ring-Fused Bisnoralcohol Derivatives as Novel Small-Molecule Antiosteoporosis Agents.

A series of heterocyclic ring-fused derivatives of bisnoralcohol (BA) were synthesized and evaluated for their inhibitory effects on RANKL-induced osteoclastogenesis. Most of these derivatives possessed potent antiosteoporosis activities in a dose-dependent manner. Among these compounds, 31 (SH442, IC50 = 0.052 µM) exhibited the highest potency, displaying 100% inhibition at 1.0 µM and 82.8% inhibition at an even lower concentration of 0.1 µM, which was much more potent than the lead compound BA (IC50 = 2.325 µM). Cytotoxicity tests suggested that the inhibitory effect of these compounds on RANKL-induced osteoclast differentiation did not result from their cytotoxicity. Mechanistic studies revealed that SH442 inhibited the expression of osteoclastogenesis-related marker genes and proteins, including TRAP, TRAF6, c-Fos, CTSK, and MMP9. Especially, SH442 could significantly attenuate bone loss of ovariectomy mouse in vivo. Therefore, these BA derivatives could be used as promising leads for the development of a new type of antiosteoporosis agent.

Effect of different retention doses of ultrasound-guided polidocanol chemical ablation for benign cystic-solid thyroid nodules.

OBJECTIVE: To assess the influence of varying retention doses of ultrasound-guided polidocanol chemical ablation for benign cystic-solid thyroid nodules. METHODS: A retrospective study was conducted from December 2019 to January 2022, including 78 patients with benign cystic-solid thyroid nodules, of which 31 received polidocanol chemical ablation alone, 23 received polidocanol chemical plus thermal ablation, and 24 received open surgery. Patients who received polidocanol chemical ablation were assigned into groups based on the retention dose of polidocanol: 0 %, 10 %, 20 %, 30 %, and 50 %. Follow-ups were done at 1, 3, 6, and 12 months postoperatively. The volume of the nodules, postoperative complications, and recurrence of the nodules were examined before treatment and during follow-up visits. RESULTS: Total operation time and intraoperative bleeding volume for patients who received ablation were substantially lower than those for patients who received open surgery (P < 0.001). Among patients in the polidocanol chemical ablation group, volume shrinkage rate of thyroid nodules in the 10 % retention dose group was significantly lower than that in the 0 % retention dose group at 1, 3, and 6 months postoperatively (P < 0.05). The 30 % retention dose group had the highest nodule shrinkage rate (98.46 ± 1.55 %) at 12 months postoperatively, which was significantly higher than that in the 50 % retention dose group (P < 0.05). Among patients in the polidocanol chemical and thermal ablation group, the volume shrinkage rate of thyroid nodules in the 10 % and 30 % retention dose groups at 1 month postoperatively was significantly lower than that in the 0 % retention dose group (P < 0.05). Although volume shrinkage rate in the 20 % retention dose group after thermal ablation was higher than that in the 0 % retention dose group, the difference was not statistically significant (P > 0.05). In terms of adverse reactions, the incidence of hoarseness and coughing was higher in the open surgery group than in the polidocanol chemical ablation and polidocanol chemical and thermal ablation groups, but there was no significant difference (P > 0.05). CONCLUSION: Chemical ablation with polidocanol was safe and effective for therapy of benign cystic-solid thyroid nodules, and the optimal retention dose may be between 20 % and 30 %. Patients with poor efficacy from chemical ablation alone can receive safe and effective treatment through thermal ablation.

Construction of a prognostic model for breast cancer based on moonlighting genes.

Breast cancer (BRCA) is a highly heterogeneous disease, with significant differences in prognosis among patients. Existing biomarkers and prognostic models have limited ability to predict BRCA prognosis. Moonlighting genes regulate tumor progression and are associated with cancer prognosis. This study aimed to construct a moonlighting gene-based prognostic model for BRCA. We obtained differentially expressed genes (DEGs) in BRCA from The Cancer Genome Atlas and intersected them with moonlighting genes from MoonProt to acquire differential moonlighting genes. GO and KEGG results showed main enrichment of these genes in the response of BRCA cells to environmental stimuli and pentose phosphate pathway. Based on moonlighting genes, we conducted drug prediction and validated results through cellular experiments. After ABCB1 knockdown, viability and proliferation of BRCA cells were significantly enhanced. Based on differential moonlighting genes, BRCA was divided into three subgroups, among which cluster2 had the highest survival rate and immunophenoscore and relatively low tumor mutation burden. TP53 had the highest mutation frequency in cluster2 and cluster3, while PIK3CA had a higher mutation frequency in cluster1, with the majority being missense mutations. Subsequently, we established an 11-gene prognostic model in the training set based on DEGs among subgroups using univariate Cox regression, LASSO regression, and multivariable Cox regression analyses. Model prognostic performance was verified in GEO, METABRIC and ICGC validation sets. In summary, this study obtained three BRCA moonlighting gene-related subtypes and constructed an 11-gene prognostic model. The 11-gene BRCA prognostic model has good predictive performance, guiding BRCA prognosis for clinical doctors.

Construction and validation of a metabolism-related prognostic model for thyroid cancer.

Metabolic reprogramming is a common pathological process of cancer. Expression of metabolism-related genes differs in thyroid cancer (TC) patients with different prognoses. This work committed to constructing a prognostic model for TC through identifying metabolism-related signatures. Expression profiles of mRNAs and clinical data of TC, were acquired from The Cancer Genome Atlas. Differential analysis was performed on mRNA expression profiles. The obtained differentially expressed genes (DEGs) were overlapped with metabolism-related genes from MSigDB database to acquire metabolism-related DEGs. Cox regression and Least Absolute Shrinkage and Selection Operator analyses were performed to ascertain feature genes and to build a prognostic model for TC. The model was evaluated comprehensively through survival curve, time-dependent receiver operating characteristic (ROC) curve, gene set enrichment analysis (GSEA), and Cox regression analyses combining varying clinical information. 7 key genes related to metabolism, including AWAT2, GGT6, ENTPD1, PAPSS2, CYP26A, ACY3 and PLA2G10, were identified, based on which a prognostic model was constructed. The survival analysis indicated that high-risk group presented shorter survival time than low-risk group. ROC curve results exhibited that AUC values of 3-year and 5-year survival of TC patients were both >0.70. Besides, GSEA on high/low-risk groups revealed that DEGs were mainly gathered in biological functions and signaling pathways linked with keratan sulfate catabolism and triglyceride catabolism. Combined with clinical information, Cox regression analyses unveiled that the 7-gene prognostic model can be an independent predictor. In conclusion, this model can effectively predict prognoses of TC patients, and also offer guidance for clinical treatment of TC.

Large depth-of-field fluorescence microscopy based on deep learning supported by Fresnel incoherent correlation holography.

Fluorescence microscopy plays an irreplaceable role in biomedicine. However, limited depth of field (DoF) of fluorescence microscopy is always an obstacle of image quality, especially when the sample is with an uneven surface or distributed in different depths. In this manuscript, we combine deep learning with Fresnel incoherent correlation holography to describe a method to obtain significant large DoF fluorescence microscopy. Firstly, the hologram is restored by the Auto-ASP method from out-of-focus to in-focus in double-spherical wave Fresnel incoherent correlation holography. Then, we use a generative adversarial network to eliminate the artifacts introduced by Auto-ASP and output the high-quality image as a result. We use fluorescent beads, USAF target and mouse brain as samples to demonstrate the large DoF of more than 400µm, which is 13 times better than that of traditional wide-field microscopy. Moreover, our method is with a simple structure, which can be easily combined with many existing fluorescence microscopic imaging technology.

Intramammary edema of invasive breast cancers on MRI T2-weighted fat suppression sequence: Correlation with molecular subtypes and clinical-pathologic prognostic factors.

OBJECTIVE: To investigate the associations between different patterns of intramammary edema on preoperative 3.0 T magnetic resonance imaging (MRI) T2-weighted fat suppression sequence and invasive breast cancer molecular subtypes and clinical-pathologic prognostic factors. METHODS: Between May 2014 and December 2020, 191 patients with invasive breast cancer who had undergone preoperative MRI and mastectomy or breast-conserving surgery were retrospectively enrolled. The relationships between different patterns of intramammary edema and invasive breast cancer molecular subtypes and clinical-pathologic features were evaluated using the Student's t-test or Mann-Whitney U test and the χ2 test or Fisher's exact test. RESULTS: Patients with luminal B (HER2 positive), HER2-enriched and triple negative breast cancers respectively had different patterns of intramammary edema (P < 0.001). There was a significant association between intramammary edema and clinical-pathologic factors, including larger tumor size, higher Ki-67 index, lymph node metastasis and lymphovascular invasion (all P < 0.001). CONCLUSIONS: Intramammary edema may provide added values of predicting molecular subtypes and clinical-pathologic prognosis, enhancing the ability to individualize the treatment of patients with invasive breast cancer.

High-definition imaging using line-illumination modulation microscopy.

The microscopic visualization of large-scale three-dimensional (3D) samples by optical microscopy requires overcoming challenges in imaging quality and speed and in big data acquisition and management. We report a line-illumination modulation (LiMo) technique for imaging thick tissues with high throughput and low background. Combining LiMo with thin tissue sectioning, we further develop a high-definition fluorescent micro-optical sectioning tomography (HD-fMOST) method that features an average signal-to-noise ratio of 110, leading to substantial improvement in neuronal morphology reconstruction. We achieve a >30-fold lossless data compression at a voxel resolution of 0.32 × 0.32 × 1.00 µm3, enabling online data storage to a USB drive or in the cloud, and high-precision (95% accuracy) brain-wide 3D cell counting in real time. These results highlight the potential of HD-fMOST to facilitate large-scale acquisition and analysis of whole-brain high-resolution datasets.

High-throughput optical sectioning via line-scanning imaging with digital structured modulation.

Optical sectioning with high-throughput, a high signal-to-noise ratio (SNR), and submicrometer resolution is crucial, but challenging, to three-dimensional visualization of large biological tissue samples. Here we propose line-scanning imaging with digital structured modulation for optical sectioning. Our method generates images with a significantly improved SNR, compared to wide-field structured illumination microscopy (WF-SIM), without residual modulation artifacts. We image a 14.5mm×11.5mm horizontal view of mouse brain tissue at a pixel resolution of 0.32µm×0.32µm in 101 s, which, compared to WF-SIM, represents a significant improvement on imaging throughput. These results provide development opportunities for high-throughput, high-resolution large-area optical imaging methods.

[Anatomic study and application of TRAM flap with partial preservation of abdominal rectus muscle in the breast reconstruction].

OBJECTIVE: To investigate the feasibility and effectiveness of transverse rectus abdominis musculocutaneous (TRAM) flap with partial preservation of abdominal rectus muscle based on the anatomic study in cadavers. METHODS: 5 adult female cadavers which provided by department of anatomy of Fujian Medical University were dissected after injection with medical red latex from the starting point of the inferior epigastric artery and superior epigastric artery. The TRAM flap with partial preservation of lateral abdominal rectus muscle were dissected for breast reconstruction. The location, route, branches and anastomosis of inferior and superior epigastric arteries were observed. Based on the anatomic study, breast reconstruction were performed in 8 cases with muscle-sparing TRAM flaps. RESULTS: The inferior epigastric artery arises from external iliac artery (9/10, 90%) or femoral artery (1/10, 10%) at the joint point between the internal third and lateral two third. There are extensive anastomoses between superior and inferior epigastric arteries above the umbilicus, mostly between the 2cm below the first tendinous intersection and umbilical level. From Sept. 2009 to Sept. 2010, 8 cases received breast reconstruction with muscle-sparing TRAM flap. The patients were followed up for 3 months to one year. Fibrosis happened in subcutaneous fat at flap IV zone in 2 cases, borderline necrosis and subcutaneous fat liquefaction occurred in some areas of flap IV zone in 2 cases, which healed after debridement. The other 4 cases healed with no complication. Except for unsatisfied shape in one case, good result achieved in 7 cases. There was no abdominal weakness, hemia or other complication. CONCLUSIONS: It is an effective and safe method in breast reconstruction with muscle-sparing TRAM flap. It is practical with comparatively short operation time and less morbidity in donor site.

[Effects of PUVA on expression of Fas and FasL in inducing apoptosis of NB4 and K562 cells].

OBJECTIVE: To explore the effects of psoralen (PSO) and long wave ultraviolet A (PUVA) on expression of Fas and FasL in apoptosis of NB4 and K562 leukemia cells. METHODS: The NB4 and K562 cells were taken as the study objects and their apoptosis ratios, ultrastructure changes and the expression of Fas and FasL were detected in order to observe the effects of PSO and UVA of wave length 360 nm on human leukemia cells. The factorial design and analysis of variance were used to analyze the interaction among the factors. RESULTS: All of PSO, UVA and PUVA could induce the apoptosis of NB4 and K562 cells, and the effects of PUVA were stronger than the other two. After treated with PUVA, the NB4 and K562 cells all showed obvious ultrastruture changes about apoptsis under the electron microscope. All of PSO, UVA and PUVA could increase the expression of Fas gene and protein, and decrease the expression of FasL gene and protein. Moreover, the effects of PUVA were stronger than the other two. CONCLUSION: PUVA can induce the apoptosis of NB4 and K562 cells and the effects are the strongest, one of the pathway of PUVA to induce apoptosis is to upregulate the expression of Fas gene and downregulate the expression of FasL gene.

[Effects of psoralen with long wave ultraviolet light on apoptosis and mitochondrial membrane potential in K562 cells].

OBJECTIVE: To study the effects of psoralen (PSO) with long wave ultraviolet light (PUVA) on apoptosis and mitochondrial membrane potential in K562 cells. METHODS: K562 cells were incubated with PSO in different concentrations (10, 20, 40 and 80 microg/ml for 24 hours) and irradiated without or with UVA (5 min). The changes in ultrastructure of the cells were observed under a transmission electron microscope. The apoptosis rates and the changes of mitochondrial membrane potentials were detected by flow cytometry. The factorial design and analysis of variance were used to analyze the interaction among the factors. RESULTS: There were obvious ultrastructure changes related to apoptosis in K562 cells after being treated with PUVA (80 microg/ml). PSO, UVA and PUVA all increased the apoptosis rates and decreased the mitochondrial membrane potentials, and the effects of PUVA were stronger than those of PSO and UVA (P<0.01). CONCLUSION: PUVA can induce the apoptosis of K562 cells and one of the pathways to the induction of apoptosis is to down-regulate the mitochondrial membrane potentials.

[Effect of psoralen plus longwave UVA inducing HL-60 cells apoptosis].

The aim of this study was to investigate the effects of the traditional Chinese medicine psoralen (PSO) plus long wave ultraviolet A (PUVA) on apoptosis in HL-60 leukemia cells and its mechanism. The effect of PUVA on HL-60 cell growth was assayed by MTT method and the changes of ultrastructure of cells were observed by electron microscopy. The apoptosis ratios, changes of mitochondrial membrane potential, expression of Fas and FasL protein and Fas and FasL mRNA were detected by FCM and fluorescent quantitation RT-PCR respectively. The expression of Caspase 8 and Caspase 3 protein were detected by immunocytochemistry (ICC). The results showed that the growth of HL-60 cells were inhibited by PUVA in time-and concentration-dependent manner through inducing cell apoptosis. When the irradiation time of long wave ultraviolet A lasted 15 minutes and the concentration of PSO was 80 microg/ml, the inhibition of HL-60 cell proliferation and apoptosis ratios reached the peak. There were obvious apoptotic ultrastructure changes and decrease of mitochondrial membrane potential in HL-60 cells after treatment with PUVA. The expression of Fas mRAN increased and expression of FasL mRNA decreased after treating with PUVA for 4 hours, and the same results of Fas, FasL expression on protein level were obtained also after treating with PUVA for 24 hours. The expression of caspase 8 and caspase 3 protein enhanced and reached the peak after treating with PUVA for 8 hours. It is concluded that the PUVA can inhibit the growth of HL-60 cells and induce apoptosis of these cells. The possible mechanism is supposed to be up-regulating Fas, down-regulating FasL levels and then activating the levels of caspase 8 and caspase 3. The decreasing of mitochondrial membrane potential may be involved in this process probably.

[Effects of psoralen plus long-wave ultraviolet-A on apoptosis of HL-60 leukemia cells].

OBJECTIVE: To explore the effects of psoralen (PSO) plus long-wave ultraviolet-A (PUVA) on apoptosis and expression of Fas ligand (FasL) in HL-60 leukemia cells. METHODS: The HL-60 cells were taken as the study objects and their apoptosis rates, ultrastructure changes and the expression of FasL were detected in order to observe the effects of PSO and ultraviolet-A (UVA) of wave length 360 nm. The factorial design and analysis of variance were used to analyze the interaction among the factors. RESULTS: PSO, UVA and PUVA all induced the apoptosis and the effects of PUVA were stronger than those of the other two. After HL-60 cells had been treated with PUVA, they all showed obvious ultrastructure changes due to apoptosis observed under the electron microscope. PSO, UVA and PUVA all decreased the expressions of FasL gene and protein. The effects of PUVA were stronger than those of the other two. CONCLUSIONS: PUVA can induce the apoptosis of HL-60 cells and the effects are stronger than those of PSO or UVA alone. The expression of FasL gene in HL-60 cells is down-regulated during the apoptosis induced by PUVA.

[Effects of PUVA on mitochondrial membrane potential in HL-60 and K562 cells].

OBJECTIVE: To study the effects of psoralen (PSO) and long wave ultraviolet light (PUVA) on mitochondrial membrane potential in HL-60 and K562 cells. METHODS: Cells were incubated with PSO in different concentrations irradiated with or without UVA. The changes of ultrastructure of cells were observed under the electron microscope. The apoptosis ratios and the changes of mitochondrial membrane potential were detected through the flow cytometry. The factorial design and analysis of variance were used to analyze the interaction among the factors. RESULTS: There were obvious ultrastructure changes about apoptosis in HL-60 and K562 cells after treated with PUVA. PSO, UVA and PUVA all increased the apoptosis ratios and decreased the mitochondrial membrane potential, and the effects of PUVA were stronger than the other two (p<0.01). CONCLUSION: PUVA can induce the apoptosis of HL-60 and K562 cells and one of the pathway to induce apoptosis is to downregulate the mitochondrial membrane potential.

[Effects of inactivated rabbit serum containing compound realgar and natural indigo tablet on cell line NB4].

OBJECTIVE: To explore the effects of inactivated rabbit serum containing compound realgar and natural indigo tablet (CRNIT) on cell line NB(4). METHODS: The experimental rabbits were taken as the provider of the animal serum, and the serum was inactivated before the experiment. The serum was divided into two groups based on whether the rabbits were given CRNIT. The concentration of arsenic in the rabbit's serum was detected by AFS-230a double path atom fluorescence photometer. The inhibition rates and apoptosis rates were regarded as the observational indexes. RESULTS: The concentration of arsenic in the inactivated rabbit serum containing and not containing the drug were (0.010 0+/-0.001 0) mg/L and (0.110 0+/-0.006 4) mg/L respectively, and the difference had statistical significance (P<0.01). The two groups of serum all had inhibitory effect on the growth of NB(4) cells depending on the drug concentration and effect time. And there were significant differences among the groups. The two groups of serum all induced the apoptosis of NB(4) with positive relations with the concentration and effect time. And there were significant differences among the groups. CONCLUSIONS: The rabbit serum containing CRNIT can obviously restrain the growth of NB(4) cells and the inhibitory effect depends on the concentration and effect time. And the rabbit serum containing CRNIT can also induce the apoptosis of NB(4) cell line and the apoptosis rates depend on the concentration and effect time.

[Growth-inhibiting effect of psoralen plus ultraviolet-A light therapy on K562 cells].

OBJECTIVE: To observe the effects of psoralen plus ultraviolet-A light (PUVA) on K562 cells and the relative mechanism. METHODS: The effects of psoralen, ultraviolet-A light and PUVA on K562 cells were assayed by monotetrazolium test (MTT). DNA content was analyzed by flow cytometry (FCM). The apoptotic rates of K562 cells treated with 40 and 80 microg/ml psoralen for 24 and 48 hours were assayed by Annexin-V-FITC/PI reagent kit on FCM respectively. The ultrastructures of apoptotic cells were observed by a transmission electron microscope (TEM). RESULTS: Either single psoralen therapy or single ultraviolet-A irradiation had inhibiting effect on K562 cells. The inhibiting effect of PUVA on K562 cells was stronger than that of the single psoralen therapy or single ultraviolet-A light irradiation (P<0.05). Apoptotic peak (AP) was detected by FCM. TEM test showed that K562 cells treated with PUVA were smaller, having condensed cell nucleus, assembled chromatin, disintegrated nucleus body and the majority of the cells appeared to be apoptotic conformation. CONCLUSION: Psoralen has inhibiting effect on K562 cells, and the effect of PUVA is more significant. It is suggested that 10 min irradiation and 40 microg/ml terminal concentration of psoralen be probably the best choice for PUVA. The inhibiting effect of PUVA is due to apoptosis.