Rapid azoospermia classification by stimulated Raman scattering and second harmonic generation microscopy.

Disease diagnosis and classification pose significant challenges due to the limited capabilities of traditional methods to obtain molecular information with spatial distribution. Optical imaging techniques, utilizing (auto)fluorescence and nonlinear optical signals, introduce new dimensions for biomarkers exploration that can improve diagnosis and classification. Nevertheless, these signals often cover only a limited number of species, impeding a comprehensive assessment of the tissue microenvironment, which is crucial for effective disease diagnosis and therapy. To address this challenge, we developed a multimodal platform, termed stimulated Raman scattering and second harmonic generation microscopy (SRASH), capable of simultaneously providing both chemical bonds and structural information of tissues. Applying SRASH imaging to azoospermia patient samples, we successfully identified lipids, protein, and collagen contrasts, unveiling molecular and structural signatures for non-obstructive azoospermia. This achievement is facilitated by LiteBlendNet-Dx (LBNet-Dx), our diagnostic algorithm, which achieved an outstanding 100% sample-level accuracy in classifying azoospermia, surpassing conventional imaging modalities. As a label-free technique, SRASH imaging eliminates the requirement for sample pre-treatment, demonstrating great potential for clinical translation and enabling molecular imaging-based diagnosis and therapy.

MiR-196b-5p activates NF-κB signaling in non-small cell lung cancer by directly targeting NFKBIA.

BACKGROUND: Our recent study found that QKI-5 regulated miRNA, miR-196b-5p, promotes non-small cell lung cancer (NSCLC) progression by directly targeting GATA6, TSPAN12 and FAS. However, the biological functions of miR-196b-5p in NSCLC progression and metastasis still remain elusive. METHODS: Cell proliferation, migration, colony formation, cell cycle assays were used to investigate cellular phenotypic changes. Quantitative real-time PCR (qRT-PCR) and western blot analyses were used to measure expressions of relative gene and protein. Interaction between QKI-5 and miR-196b-5p was determined by RNA immunoprecipitation (RIP) assay. Luciferase reporter assay was used to determine direct binding between miR-196b-5p and NFKBIA 3'-UTR. ELISA assay was used to measure secreted IL6 proteins. Mice xenograft model was used to assess the functions of NFKBIA on in vivo tumor growth. RESULTS: We demonstrated that the miR-196b-5p facilitates lung cancer cell proliferation, migration, colony formation, and cell cycle by directly targeting NFKBIA, a negative regulator of NFκB signaling. Knocking down NFKBIA increases IL6 mediated phosphorylation of signal transducer and activator of transcription 3 (STAT3) to promote lung cancer cell growth by activating NFκB signaling. The expression of NFKBIA was significantly downregulated in NSCLC tissue samples, and was negatively correlated with the expression miR-196b-5p. In addition, we found that downregulated QKI-5 expression was associated with the elevated miR-224 expression in NSCLC. CONCLUSIONS: Our findings indicated that the miR-224/QKI-5/miR-196b-5p/NFKBIA signaling pathway might play important functions in the progression of NSCLC, and suggested that targeting this pathway might be an effective therapeutic strategy in treating NSCLC.

miR-31/QKI-5 axis facilitates cell cycle progression of non-small-cell lung cancer cells by interacting and regulating p21 and CDK4/6 expressions.

BACKGROUND: RNA-binding protein Quaking-5 (QKI-5), a major isoform of QKIs, inhibits tumor progression in non-small cell lung cancer (NSCLC). However, the underlying molecular mechanisms of QKI-5 in the cell cycle of NSCLC are still largely unknown. METHODS: MTT, flow cytometry, and colony formation assays were used to investigate cellular phenotypic changes. Mice xenograft model was used to evaluate the antitumor activities of QKI-5. Co-immunoprecipitation, RNA immunoprecipitation (RIP), and RIP sequencing were used to investigate protein-protein interaction and protein-mRNA interaction. RESULTS: The QKI-5 expression was downregulated in NSCLC tissues compared with that in paired normal adjacent lung tissues. Overexpression of QKI-5 inhibited NSCLC cell proliferative and colony forming ability. In addition, QKI-5 induced cell cycle arrest at G0/G1 phase through upregulating p21Waf1/Cip1 (p21) expression and downregulating cyclin D1, cyclin-dependent kinase 4 (CDK4), and CDK6 expressions. Further analyses showed that QKI-5 interacts with p21 protein and CDK4, CDK6 mRNAs, suggesting a critical function of QKI-5 in cell cycle regulation. In agreement with in vitro study, the mouse xenograft models validated tumor suppressive functions of QKI-5 in vivo through altering cell cycle G1-phase-associated proteins. Moreover, we demonstrated that QKI-5 is a direct target of miR-31. The QKI-5 expression was anticorrelated with the miR-31 expression in NSCLC patient samples. CONCLUSION: Our results suggest that the miR-31/QKI-5/p21-CDK4-CDK6 axis might have critical functions in the progression of NSCLC, and targeting this axis could serve as a potential therapeutic strategy for NSCLC.

Association between vitamin D serum levels and insulin resistance assessed by HOMA-IR among non-diabetic adults in the United States: Results from NHANES 2007-2014.

Insulin resistance, a pathological response to insulin hormone in insulin-dependent cells, is characterized by the presence of high glucose and insulin concentrations. The homeostasis model of insulin resistance (HOMA-IR) is one of the most used indexes to estimate insulin resistance by assessing the fasting glucose and insulin levels. An association was observed between vitamin D levels and insulin resistance, which varied in different ethnic groups, and there is some evidence that vitamin D supplementation could contribute to the improvement of insulin resistance. This study assessed the association between 25-hydroxyvitamin D (25[OH]D) concentration and HOMA-IR in American adults aged 20 years and older, without diabetes and other chronic diseases that can influence insulin resistance. The data from the National Health and Nutrition Examination Survey (NHANES) 2007-2014 were used by exploiting the free and publicly-accessible web datasets. Linear regression models were performed to evaluate the association between serum 25(OH)D concentration and HOMA-IR, and a negative association was observed, which remained significant following the adjustment for age, gender, race/ethnicity, education, body mass index (BMI), physical activity, the season of examination, current smoking, hypertension, the use of drugs which can influence insulin resistance, serum bicarbonates, triglycerides, and calcium and phosphorus levels. Only in non-Hispanic Blacks was this inverse association between vitamin D and HOMA-IR not observed in the fully adjusted model. Further studies are needed to explain the mechanisms of the observed ethnic/racial differences in the association of vitamin D levels with HOMA-IR.

Frontiers of MicroRNA Signature in Non-small Cell Lung Cancer.

Lung cancer is the leading cause of cancer-related deaths worldwide and non-small cell lung cancer (NSCLC) accounts for more than 80% of all lung cancer cases. Recent advancements in diagnostic tools, surgical treatments, chemotherapies, and molecular targeted therapies that improved the therapeutic efficacy in NSCLC. However, the 5-years relative survival rate of NSCLC is only about 20% due to the inadequate screening methods and late onset of clinical symptoms. Dysregulation of microRNAs (miRNAs) was frequently observed in NSCLC and closely associated with NSCLC development, progression, and metastasis through regulating their target genes. In this review, we provide an updated overview of aberrant miRNA signature in NSCLC, and discuss the possibility of miRNAs becoming a diagnostic and therapeutic tool. We also discuss the possible causes of dysregulated miRNAs in NSCLC.

Dissecting miRNA signature in colorectal cancer progression and metastasis.

Colorectal cancer (CRC) is the third most common cancer and leading cause of cancer related deaths worldwide. Despite recent advancements in surgical and molecular targeted therapies that improved the therapeutic efficacy in CRC, the 5 years survival rate of CRC patients still remains frustratingly poor. Accumulated evidences indicate that microRNAs (miRNAs) play a crucial role in the progression and metastasis of CRC. Dysregulated miRNAs are closely associated with cancerous phenotypes (e.g. enhanced proliferative and invasive ability, evasion of apoptosis, cell cycle aberration, and promotion of angiogenesis) by regulating their target genes. In this review, we provide an updated overview of tumor suppressive and oncogenic miRNAs, circulatory miRNAs, and the possible causes of dysregulated miRNAs in CRC. In addition, we discuss the important functions of miRNAs in drug resistance of CRC.

miR-196b-5p-mediated downregulation of FAS promotes NSCLC progression by activating IL6-STAT3 signaling.

Our recent study demonstrated that the QKI-5 regulated miRNA, miR-196b-5p, and it functions as an onco-microRNA in non-small cell lung cancer (NSCLC) by directly targeting GATA6 and TSPAN12. However, the role of miR-196b-5p in NSCLC progression and metastasis still remains unclear. We found that miR-196b-5p promotes lung cancer cell proliferation and colony formation by directly targeting tumor suppressor, FAS. The expression of FAS was significantly downregulated in NSCLC tissue samples and was negatively correlated with the miR-196b-5p expression. Knocking down FAS activates NFkB signaling and subsequent IL6 secretion, resulting in phosphorylation of signal transducer and activator of transcription 3 (STAT3) to promote lung cancer cell growth. Our findings indicated that miR-196b-5p might exhibit novel oncogenic function by FAS-mediated STAT3 activation in NSCLC, and suggested that targeting the miR-196b-5p/FAS/NFkB/IL6/STAT3 pathway might be a promising therapeutic strategy in treating NSCLC.

miR-196b-5p-mediated downregulation of TSPAN12 and GATA6 promotes tumor progression in non-small cell lung cancer.

Lung cancer is the leading cause of cancer-related deaths worldwide and non-small cell lung cancer (NSCLC) accounts for over 80% of lung cancer cases. The RNA binding protein, QKI, belongs to the STAR family and plays tumor-suppressive functions in NSCLC. QKI-5 is a major isoform of QKIs and is predominantly expressed in NSCLC. However, the underlying mechanisms of QKI-5 in NSCLC progression remain unclear. We found that QKI-5 regulated microRNA (miRNA), miR-196b-5p, and its expression was significantly up-regulated in NSCLC tissues. Up-regulated miR-196b-5p promotes lung cancer cell migration, proliferation, and cell cycle through directly targeting the tumor suppressors, GATA6 and TSPAN12. Both GATA6 and TSPAN12 expressions were down-regulated in NSCLC patient tissue samples and were negatively correlated with miR-196b-5p expression. Mouse xenograft models demonstrated that miR-196b-5p functions as a potent onco-miRNA, whereas TSPAN12 functions as a tumor suppressor in NSCLC in vivo. QKI-5 bound to miR-196b-5p and influenced its stability, resulting in up-regulated miR-196b-5p expression in NSCLC. Further analysis showed that hypomethylation in the promoter region enhanced miR-196b-5p expression in NSCLC. Our findings indicate that QKI-5 may exhibit novel anticancer mechanisms by regulating miRNA in NSCLC, and targeting the QKI5∼miR-196b-5p∼GATA6/TSPAN12 pathway may enable effectively treating some NSCLCs.

[Short-term effectiveness of acetabular reconstruction with three-dimensional printed trabecular metal pads in hip revision].

OBJECTIVE: To investigate the short-term effectiveness of three-dimensional (3D) printed trabecular metal pads for Paprosky type Ⅲ acetabular defect in hip revision. METHODS: Between August 2014 and December 2015, the 3D printed trabecular metal pads were used to repair Paprosky type Ⅲ acetabular defects and reconstruct the annular structure of the acetabulums in 5 cases of primary hip revision. There were 3 males and 2 females, aged from 50 to 72 years, with an average age of 66 years. The time from initial replacement to revision was 10 to 18 years, with an average of 14.4 years. The types of prostheses in primary replacement were non-cemented in 3 cases and cemented in 2 cases. The types of acetabular bone defects were Paprosky type ⅢA in 3 cases and Paprosky type ⅢB in 2 cases. The preoperative Harris score was 34.23±11.67. The height of rotation center of affected hip was (38.17±8.87) mm and the horizontal position was (35.62±9.12) mm. RESULTS: The operation time was 120-180 minutes, with an average of 142 minutes. The intraoperative bleeding volume was 800-1 700 mL, with an average of 1 100 mL. Five patients were followed up 18-24 months (mean, 21 months). At last follow-up, the Harris score was 79.82±8.70, which was significantly higher than that before operation (t=16.991, P=0.000). At 1 week after operation, the abduction angle of acetabular cup was 38-42° (mean, 39.4 °) and the anteversion angle was 13-18 ° (mean, 14.6°). The height and horizontal position of rotation center of affected hip were (22.08±8.33) mm and (29.03±6.28) mm, respectively, showing significant differences when compared with those before operation (P<0.05); there was no significant difference when compared with those of healthy hip [(28.62±7.73), (27.29±4.22) mm] (P>0.05). During the follow-up, there was no complication such as prosthesis loosening, dislocation, or periprosthetic fracture. CONCLUSION: In hip revision, 3D printed trabecular metal pads can repair Paprosky type Ⅲ acetabular defect, reconstruct the structure of acetabulum, provide a stable supporting structure for the acetabular cup, reconstruct the relatively normal rotation center of the hip joint, avoid iatrogenic bone loss, and achieve satisfactory functional recovery of the hip. The long-term effectiveness needs further follow-up.

Curcumin derivative WZ35 inhibits tumor cell growth via ROS-YAP-JNK signaling pathway in breast cancer.

BACKGROUND: Breast cancer is the most prevalent cancer among women worldwide. WZ35, an analog of curcumin, has been demonstrated to remarkably improve the pharmacokinetic profiles in vivo compared with curcumin. WZ35 exhibits promising antitumor activity in gastric cancer, HCC, colon cancer. However, antitumor effects of WZ35 in breast cancer and its underlying molecular mechanisms remain unclear. METHODS: CCK8, Flow cytometry and transwell assays were used to measure cell proliferation, cell cycle arrest, apoptosis, cell migration and invasion. We constructed xenograft mouse model and lung metastasis model to assess the antitumor activities of WZ35 in vivo. To explore the underlying molecular mechanisms of WZ35, we performed a series of overexpression and knockdown experiments. The cellular oxygen consumption rates (OCRs) was measured to assess mitochondrial dysfunction. RESULTS: We found that treatment of breast cancer cells with WZ35 exerts stronger anti-tumor activities than curcumin both in vitro and in vivo. Mechanistically, our research showed that WZ35 induced reactive oxygen species (ROS) generation and subsequent YAP mediated JNK activation in breast cancer cells. Abrogation of ROS production markedly attenuated WZ35 induced anti-tumor activities as well as YAP and JNK activation. In addition, ROS mediated YAP and JNK activation induced mitochondrial dysfunction in breast cancer cells. CONCLUSION: Our study showed that novel anti-cancer mechanisms of WZ35 in breast cancer cells and ROS-YAP-JNK pathway might be a potential therapeutic target for the treatment of breast cancer patients.

Alantolactone promotes ER stress-mediated apoptosis by inhibition of TrxR1 in triple-negative breast cancer cell lines and in a mouse model.

Triple-negative breast cancer (TNBC) is a subtype of breast cancer with poor clinical outcome and currently no effective targeted therapies are available. Alantolactone (ATL), a sesquiterpene lactone, has been shown to have potential anti-tumour activity against various cancer cells. However, the underlying mechanism and therapeutic effect of ATL in the TNBC are largely unknown. In the present study, we found that ATL suppresses TNBC cell viability by reactive oxygen species (ROS) accumulation and subsequent ROS-dependent endoplasmic reticulum (ER) stress both in vitro and in vivo. Thioredoxin reductase 1 (TrxR1) expression and activity of were significantly up-regulated in the TNBC tissue specimens compare to the normal adjacent tissues. Further analyses showed that ATL inhibits the activity of TrxR1 both in vitro and in vivo in TNBC and knockdown of TrxR1 in TNBC cells sensitized ATL-induced cell apoptosis and ROS increase. These results will provide pre-clinical evidences that ATL could be a potential therapeutic agent against TNBC by promoting ROS-ER stress-mediated apoptosis through partly targeting TrxR1.

Clinical evaluation following the use of mineralized collagen graft for bone defects in revision total hip arthroplasty.

Revision total hip arthroplasty (THA) with massive bone loss has been a real challenge for orthopaedic surgeons. Here we describe an approach using mineralized collagen (MC) graft to reconstruct acetabulum and femur with massive bone defects. We identified 89 patients suffering acetabular or femoral bone defects after primary THA, who required revision THA for this study. During the surgery, MC was applied to reconstruct both the acetabular and femoral defects. Harris hip score was used to evaluate hip function while radiographs were taken to estimate bone formation in the defect regions. The average follow-up period was 33.6 ± 2.4 months. None of the components needed re-revised. Mean Harris hip scores were 42.5 ± 3.5 before operation, 75.2 ± 4.0 at 10th month and 95.0 ± 3.6 at the final follow-up. There were no instances of deep infection, severe venous thrombosis or nerve palsy. The present study demonstrated that MC graft can serve as a promising option for revision THA with massive bone deficiency. Meanwhile, extended follow-up is needed to further prove its long-term performance.

Core decompression and implantation of calcium phosphate cement/Danshen drug delivery system for treating ischemic necrosis of femoral head at Stages I, II and III of antigen reactive cell opsonization.

OBJECTIVE: To introduce a new method using calcium phosphate cement/Danshen drug delivery system for treating ischemic necrosis of the femoral head and evaluate its curative effect. METHODS: From May 2000 to June 2005, 48 adult patients (54 hips) with ischemic necrosis of the femoral head at Stages I, II and III of antigen reactive cell opsonization (ARCO) were treated with implantation of calcium phosphate cement/Danshen drug delivery system in the involved femoral head. The operation consisted of removal of the necrotic bone under weight-loading cartilage and the implantation of phosphate cement/Danshen drug delivery system, and all manipulations were made percutaneously through a bone tunnel in the trochanter. The functions of the hip joint were evaluated and X-ray films were taken preoperatively and postoperatively. RESULTS: Postoperative follow-up was 45.5 months on average, ranging from 27 to 78 months. According to the evaluation criterion of "Dandong 1995" for therapeutic effect of adult ischemic necrosis of the femoral head, the therapeutic effects were excellent in 33 hips, good in 17, fair in 3 and poor in 1, with the excellent and good rate of 92.6%. CONCLUSIONS: This method is relatively simple with little invasion. It not only improves the microcirculation of the femoral head by local application of traditional Chinese medicine, but also provides mechanic buttress in the weight-loading area to prevent collapse during repairing, which is beneficial to repair and reconstruction of femoral head. It may be a choice of minimal invasion surgery for ischemic necrosis of the femoral head at Stages I, II and III of ARCO.

[Clinical study on the early reconstruction of complex acetabular fractures].

OBJECTIVE: To explore the best opportunity of structure reconstruction for complex acetabular fracture in order to provide the accurate time for clinical operative treatment. METHODS: Complex acetabular fracture patients were divided into experimental group (93 cases, 96 hips) and control group (98 cases, 101 hips) randomly according to the operative time. The operation of patients were done respectively at the 7th, 10th day after injury in experimental group and control group. The operative time, the excellent and good rate of reduction,the postoperative complications,the joint function (ache to walk,joint activity),the SF-36 were evaluated. RESULTS: (1) The operative time of experimental group was obviously shorter than control group according to different fracture classification. (2) According to the standard of Matta' joint function and X-ray, the experimental group was better than control group. (3) The excellent and good rate of reduction in experimental group was obviously higher than control group, according to CT scan before operation and after operation. CONCLUSION: The early structure reconstruction of complex acetabular fracture can obviously decrease operative time and complications, increase the rate of operative reduction.

[Implantation of calcium phosphate cement/Danshen drug delivery system for avascular necrosis of femoral head].

OBJECTIVE: To introduce a new method using calcium phosphate cement/Danshen drug delivery system for avascular necrosis of femoral head and to evaluate its clinical outcome. METHODS: From May 2000 to June 2005, 48 patients (54 hips) with avascular necrosis of femoral head were treated with calcium phosphate cement/Danshen drug delivery system implantation in the involved femoral head. There were 32 males (36 hips) and 16 females (18 hips) with an average age of 38.7 years (26-62 years). Twenty-one cases had the history of drinking or smoking, 15 cases had the history of receivihg hormonotherapy and 2 had the history of injury in hip joint. The disease course was 2-32 months. According to standard of Association Research Circulation Osseous (ARCO) staging, 9 hips were classified as stage I, 31 as stage II and 14 as stage III The operation consisted of removal of necrotic bone under weight-loading cartilage and the implantation of calcium phosphate cement/Danshen drug delivery system, all manipulations were done through a bone tunnel in trochanter. The function of hip joint were evaluated and X-ray films were taken pre- and post-operatively. RESULTS: No phlebothrombosis of leg and foreign body action occurred in all cases, and incision healed by first intention. The postoperative follow-up averaged 42.5 months, ranging from 22 to 73 months. According to the evaluation criterion of Dandong 1995 for adult avascular necrosis of femoral head, the results were excellent in 33 hips, good in 17, fair in 3 and poor in 1, the excellent and good rate was 92.6%. CONCLUSION: This method is relatively simple with less invasion, it not only improves the microcirculation of femoral head by local application of traditional Chinese medicine, but also provide mechanic buttress in the weight-loaded area, which is beneficial to repair and reconstruction of femoral head. It may be a choice of minimally invasion surgery for femoral head necrosis.

[Physicochemical properties and drug release rate of calcium phosphate cement containing Danshen composite injection].

OBJECTIVE: To investigate the physicochemical properties of the calcium phosphate cement (CPC) containing Danshen composite injection and its drug release rate. METHODS: This experiment included 4 groups and each group contained 6 specimens. CPC (2 g) was mixed with the setting solution that served as the control group; 0.1, 0.5 and 1.0 ml of Danshen composites injection (concentration, 1 000 mg/ml; pH, 7.35) were respectively added to CPC (2 g), which were used as the experimental groups 1, 2 and 3. The resulting specimens were investigated by the X-ray diffraction (XRD), the fourier transformed infrared spectroscopy (FTIR), and the scanning electron microscope (SEM). RESULTS: The XRD analysis showed that the control group had a typical diffraction pattern of the hydroxypatite (HAP), which was consistent with the standard pattern of HAP. When more Danshen was added in the experimental groups, the diffraction peaks of HAP gradually decreased; when the diffraction angle 2theta was about 25.92 degrees, the HAP peaks disappeared. Based on the FTIR analysis, with an increase of the drug concentration, the absorption peak of the hydroxy groups decreased. The SEM showed that the size of the CPC particle was related to the drug concentration; with an increase of the drug concentration, the CPC particle increased in number, resulting in an increasing trend of coacervation. The elution test showed that the drug-release rate and capacity varied with the different concentrations of Danshen. The initial release rate was relatively great, but after 96 hours the rate slowed down, lasting for a long time. CONCLUSION: The physicochemical properties of CPC do not change when a proper dose (0.1 ml/2 g) of Danshen is added to CPC. The Danshen composite can be effectively released from CPC, and so CPC can be used as an ideal drug-delivery carrier for Danshen composite.