PKD1 mutant clones within cirrhotic livers inhibit steatohepatitis without promoting cancer.

Somatic mutations in non-malignant tissues are selected for because they confer increased clonal fitness. However, it is uncertain whether these clones can benefit organ health. Here, ultra-deep targeted sequencing of 150 liver samples from 30 chronic liver disease patients revealed recurrent somatic mutations. PKD1 mutations were observed in 30% of patients, whereas they were only detected in 1.3% of hepatocellular carcinomas (HCCs). To interrogate tumor suppressor functionality, we perturbed PKD1 in two HCC cell lines and six in vivo models, in some cases showing that PKD1 loss protected against HCC, but in most cases showing no impact. However, Pkd1 haploinsufficiency accelerated regeneration after partial hepatectomy. We tested Pkd1 in fatty liver disease, showing that Pkd1 loss was protective against steatosis and glucose intolerance. Mechanistically, Pkd1 loss selectively increased mTOR signaling without SREBP-1c activation. In summary, PKD1 mutations exert adaptive functionality on the organ level without increasing transformation risk.

Optimized sand tube irrigation combined with nitrogen application improves jujube yield as well as water and nitrogen use efficiencies in an arid desert region of Northwest China.

Efficient water-saving irrigation techniques and appropriate nitrogen (N) application are keys to solving the problems of water scarcity and irrational fertilization in jujube cultivation. In this study, first, the effects of sand tube irrigation (STI) on surface and subsurface wetted characteristics were investigated using in-situ infiltration tests in a jujube garden. Compared with surface drip irrigation (SD), STI reduced surface wetted area by 57.4% and wetted perimeter of the surface wetted circle by 37.1% and increased subsurface maximum infiltration distance of wetting front by 64.9%. At the optimal sand tube depth of 20 cm, surface wetted area of the surface wetted circle decreased by 65.4% and maximum infiltration distance of the wetting front increased by 70.9%, compared with SD. Two-year field experiments then investigated the effects of STI and SD on soil water storage, jujube leaf chlorophyll, net photosynthetic rate, actual water consumption, fruit yield, and water (WUE) and N (NUE) use efficiencies at four levels of N (pure nitrogen: N1, 0; N2, 286 kg ha-1; N3, 381 kg ha-1; N4, 476 kg ha-1) at the same irrigation amount (45 mm irrigation-1, total of 8). Compared with SD, STI increased soil water storage 18.0% (2021) and 15.6% (2022) during the entire growth period and also chlorophyll content, nitrogen balance index, and net photosynthetic rate, with both increasing and then decreasing with increasing N. Compared with SD, STI increased yields by 39.1% and 36.5% and WUE by 44.3% and 39.7% in 2021 and 2022, respectively. Nitrogen use efficiency was 2.5 (2021) and 1.6 (2022) times higher with STI than with SD. STI combined with N3 had the highest yield, WUE, NUE, and net income and is thus recommended as the optimal water-N combination. In conclusion, STI combined with appropriate N application can be an effective water-saving irrigation technology alternative to SD in jujube cultivation in arid areas.

Chemokine receptors differentially expressed by race category and molecular subtype in the breast cancer TCGA cohort.

Racial disparities in mortality due to metastasis remain significant among breast cancer patients. Chemokine receptors contribute to breast tumors and metastatic outcome. We explored for significant differences in chemokine receptor expression in breast tumors from Black, Asian, and White patients in The Cancer Genome Atlas. We show that despite sharing the same molecular subtype, expression of the chemokine receptors ACKR1, CCR3, CCR6, CCRL1, CCRL2, CXCR1, CXCR2, CXCR4, CXCR6, and CXC3CR1 was significantly different depending on racial group. For patients with triple negative breast cancer, CCR3 was higher in Black versus White and CCRL2 was higher in Asian versus White. In luminal A tumors, ACKR1 was lower in Asian versus White, CCR3 was higher in Black versus White, and CCR6 and CXC3CR1 were lower in Black versus White. In luminal B tumors, CCRL2 was lower in Black versus White, CXCR1 and CXC3CR1 were lower in Asian versus White, and CXCR2 was lower in Black and Asian versus White. In HER2 enriched tumors, CCR3 was higher in Black versus White and CXCR4 lower in Asian versus White. CCR3, CCR6, and CXCR6 associated with worse patient survival. These findings can inform improved treatment strategies to decrease racial disparities in breast cancer burden.

Novel Competitive Chemiluminescence DNA Assay Based on Fe3O4@SiO2@Au-Functionalized Magnetic Nanoparticles for Sensitive Detection of p53 Tumor Suppressor Gene.

A simple, rapid response time and ultrahigh sensitive chemiluminescence (CL) DNA assay based on Fe3O4@SiO2@Au-functionalized magnetic nanoparticles (Au-MNPs) was developed for detection of p53 tumor suppressor gene. In this study, 2',6'-dimethylcarbonylphenyl-10-sulfopropyl acridinium-9-carboxylate 4'-NHS ester (NSP-DMAE-NHS), as a new kind of highly efficient luminescence reagent, was immobilized on the complementary sequence of the wild-type p53 (ssDNA) to improve the detection sensitivity. The optimal concentration of ssDNA-(NSP-DMAE-NHS) conjugates mixed with the wild-type p53 (wtp53) samples respectively. Then, the wtp53-Au-MNPs conjugates were added to continue the competitive reaction in the above solution. Subsequently, the Au-MNPs separated under magnetic field, measured by a homemade luminescent measurement system. Under optimal conditions, the method exhibited ultrasensitive sensitivity with a detection limit of 0.001 ng mL-1 (0.16 pM), a wide range of liner response from 0.001 ng mL-1~6.6 µg mL-1. Therefore, the immunomagnetic nanocomposites-based detection strategy was rapid, low-cost, and highly sensitive that can be easily extended to the early diagnosis of cancer development and monitoring of patient therapy.

A morphometric study of the middle and lower cervical vertebral endplates and their components.

Cervical disc arthroplasty is a common method of treating cervical degenerative disease. However, the footprints of most prosthesis dimensions are obtained from data of Caucasian individuals. Besides, there is a large discrepancy between footprints of currently available cervical disc prostheses and anatomic dimensions of cervical endplates. We aimed to detail the three-dimensional (3D) anatomic morphology of the subaxial cervical vertebral endplate, utilizing high-precision, high-resolution scanning equipment, and provide a theoretical basis for designing appropriate disc prostheses for Chinese patients.A total of 138 cervical vertebral endplates were studied. Each endplate was digitized using a non-contact optical 3D range scanning system and then reconstructed to quantify diameters and surface area for the whole endplate and its components (central endplate and epiphyseal rim). The whole endplate and mid-plane concavity depth were measured.There is marked morphologic asymmetry, in that the cranial endplate is more concave than the corresponding caudal endplate, with endplate concavity depths of 2.04 and 0.69 mm, respectively. For the caudal endplates, the endplate concavity apex locations were always located in the posterior portion (81.42%), while in cranial endplates relatively even. The central endplate was approximately 60% of the area of the whole endplate and the anterior section of the ring was the widest. From C3/4 down to C6/7 discs, the vertebral endplate gradually became more elliptical. Chinese cervical endplate anatomic sizes are generally smaller than that of Caucasians. Although Korean and Chinese individuals both belong to the Asian population subgroup, the majority of anatomic dimensions differ. Singaporean cervical endplate morphology is very similar to that of Chinese patients.We performed a comprehensive and accurate quantitative description of the cervical endplate, which provide references to shape and profile an artificial cervical disc without sacrificing valuable bone stock. To design a device with footprint as large as possible to distribute the axial load, we suggest that additional attention should be paid to the marginal rim. It is essential to specifically design appropriate disc prosthesis for Chinese patients. To fit the morphologic and biomechanical variations, we also propose that the disc prostheses for different vertebral segments should be separately designed.

Quantitative morphometric study of the subaxial cervical vertebrae end plate.

BACKGROUND CONTEXT: Cervical disc arthroplasty has been gradually adopted as an alternative for the treatment of cervical degenerative disease. However, there is a large discrepancy between footprints of currently available cervical disc prostheses and anatomic dimensions of cervical end plates. PURPOSE: This study aimed to accurately and comprehensively quantify the three-dimensional (3D) anatomic morphology of the cervical vertebral end plate and provide a theoretical basis for designing appropriate disc prostheses. Moreover, we introduced a novel geometric and mechanical model for 3D reconstruction techniques of the cervical end plate. STUDY DESIGN: A descriptive study of the geometry of the middle and lower cervical vertebral end plates in cadaveric spines was carried out. METHODS: A total of 138 cervical vertebral end plates were digitized using an optical 3D range scanning system, and then each end plate was reconstructed using the digitized image. For each end plate, the morphologic characteristics of six surface curves and the end plate concavity depth were symmetrically chosen and depicted. RESULTS: The cranial end plates (relative to the disc) were concave and the caudal end plates were relatively flat at all disc levels, with mean concavity depths of 2.04 and 0.69 mm, respectively. For the caudal end plates, the end plate concavity apex was most often (81.42%) located in the posterior portion, whereas in the cranial end plates, the distribution was relatively even. For the sagittal curves, the foremost point and the rearmost point on the middle curve had a more forward position than those in the left curve and the right curve. Regarding the frontal plane curves, the length of the middle curve was longer than that of the anterior curve and posterior curve. For the cranial end plate, the maximal mean depth was the middle curve, whereas for the caudal end plate, the maximum depth was the posterior curve. CONCLUSIONS: There is marked morphologic asymmetry, in that the cranial end plate is more concave than the corresponding caudal end plate. In the sagittal plane, the caudal end plates are aerofoil-shaped, whereas the cranial end plates are arc-shaped. In the transverse plane, the end plates are kidney-shaped. These morphologic characteristics of cervical vertebral end plates should be taken into consideration when designing cervical devices, such as artificial discs.

Direct competitive chemiluminescence immunoassays based on gold-coated magnetic particles for detection of chloramphenicol.

Direct competitive chemiluminescence immunoassays (CLIA) based on gold-coated magnetic nanospheres (Au-MNPs) were developed for rapid analysis of chloramphenicol (CAP). The Au-MNPs were modified with carboxyl groups and amino groups by 11-mercaptoundecanoic acid (MUA) and cysteamine respectively, and then were respectively conjugated with CAP base and CAP succinate via an activating reaction using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS). NSP-DMAE-NHS, a new and effective luminescence reagent, was employed to label anti-CAP antibody (mAb) as a tracer in direct CLIA for CAP detection using a 'homemade' luminescent measurement system that was set up with a photomultiplier tube (PMT) and a photon counting unit linked to a computer. The sensitivities and limits of detection (LODs) of the two methods were obtained and compared according to the inhibition curves. The 50% inhibition concentration (IC50 ) values of the two methods were about 0.044 ng/mL and 0.072 ng/mL respectively and LODs were approximately 0.001 ng/mL and 0.006 ng/mL respectively. To our knowledge, they were much more sensitive than any traditional enzyme-linked immunosorbent assay (ELISA) ever reported. Moreover, the new luminescence reagent NSP-DMAE-NHS is much more sensitive and stable than luminol and its derivatives, contributing to the sensitivity enhancement.

Clenbuterol Assay by Spectral Imaging Surface Plasmon Resonance Biosensor System.

To prevent illegal use of clenbuterol and for quality control in the food industry, more efficient and reliable methods for clenbuterol detection are needed. In this study, clenbuterol was detected using a spectral imaging surface plasmon resonance sensor system via two inhibition methods: (1) the target site compensation method, in which anti-clenbuterol antibody was immobilized on the sensor chip as a bioprobe and (2) the solution competition method in which a clenbuterol-BSA conjugate was immobilized on the sensor chip as the bioprobe. The detectable clenbuterol concentration ranged between 6.25 and 100 µg/mL for both methods. The clenbuterol limit of detection for the target site compensation method and solution competition method are estimated to be 6.7 and 4.5 µg/mL, respectively. The proposed methods were successfully applied to the detection of clenbuterol molecules and were found to have high specificity and high-throughput and were label free and operationally convenient.

Posterior atlantoaxial fixation: a review of all techniques.

BACKGROUND CONTEXT: Posterior atlantoaxial fixation is an effective treatment for atlantoaxial instability. Great advancements on posterior atlantoaxial fixation techniques have been made in the past decades. However, there is no article reviewing all the posterior atlantoaxial fixation techniques yet. PURPOSE: The aim was to review the evolution and advancements of posterior atlantoaxial fixation. STUDY DESIGN: This was a literature review. METHODS: The application of all posterior fixation techniques in atlantoaxial stabilization, including wiring techniques, interlaminar clamp fixation, transarticular fixation, screw-plate systems, screw-rod systems, and hook-screw systems, are reviewed and discussed. Recent advancements on the novel technique of atlantoaxial fixation are described. The combination of the C1 and C2 screws in screw-rod systems are described in detail. RESULTS: All fixation techniques are useful. The screw-rod system appears to be the most popular approach. However, many novel or modified fixation methods have been introduced in recent years. CONCLUSIONS: Great advancements on posterior atlantoaxial fixation techniques have been made in the past decades. The wiring technique and interlaminar clamps technique have fallen out of favor because of the development of newer and superior fixation techniques. The C1-C2 transarticular screw technique may remain the gold standard for atlantoaxial fusion, whereas screw-rod systems, especially the C1 pedicle screw combined with C2 pedicle/pars screw fixation, have become the most popular fixation techniques. Hook-screw systems are alternatives for atlantoaxial fixation.

[Incidences of C5 nerve palsy after multi-segmental cervical decompression through different approaches].

OBJECTIVE: To investigate the incidence of C5 nerve root palsy after multi-segmental cervical decompression through different approaches. METHODS: This study was conducted among 375 patients undergoing multi-segmental cervical decompression in anterior corpectomy and fusion fixation, anterior cervical corpectomy and fusion fixation + posterior decompression and fusion fixation, posterior cervical laminectomy decompression, fusion and internal fixation, and posterior laminoplasty and fusion groups. The exclusion criteria included lack of follow-up data, spinal cord injury preventing preoperative or postoperative motor testing, or surgery not involving the C5 level. The incidence of C5 palsy was determined and the potential risk factors C5 palsy were analyzed including age, sex, revision surgery, preoperative weakness, diabetes, smoking, number of levels decompressed, and a history of previous upper extremity surgery. RESULTS: Of the 375 patients, 60 patients were excluded and the data of 315 patients were analyzed, including 146 women and 169 men with a mean age of 57.7 years (range 39-72 years). The overall incidence of C5 nerve palsy was 6.03% (19/315) in these patients; in the subgroups receiving different surgeries, the incidence was 8.62% in the cervical road laminectomy and fusion fixation group, 7.79% in the anterior cervical corpectomy and fusion fixation + posterior decompression and fusion and internal fixation, 4.68% in the anterior corpectomy and fusion fixation group, and 3.85% in the posterior laminoplasty and fusion group. No significant difference was found in the incidences among the subgroups, but men were more likely than women to develop cervical nerve root palsy (8.28% vs 3.42%, P<0.05). CONCLUSION: The overall incidence of C5 nerve palsy following postoperative cervical spinal decompression was 6.03% in our cohort. The incidence of C5 nerve palsy did not differ significantly following different cervical decompression surgeries, but the incidence was the highest in the posterior cervical laminectomy and fusion and internal fixation group.

Spectral surface plasmon resonance imaging for the detection of clenbuterol via three-dimensional immobilization of bioprobes.

A method of immobilizing clenbuterol (CLEN) on the sensor chip for spectral surface plasmon resonance imaging (SPRi) was experimentally investigated. The bioprobes on the sensor chip were prepared by immobilizing bovine serum albumin (BSA) protein and conjugating CLEN molecules to BSA, which provides more active points and free orientations for specific binding. The calibration curve showed that the wavelength resonance shift decreased as the concentration of CLEN analyte increased, consistent with the inhibition principle. The limit of detection (LOD) was estimated to be 6.32 µg/ml. This method proved to be highly specific, high throughput, label free, and operationally convenient.

Ginsenoside Rd attenuates mitochondrial permeability transition and cytochrome C release in isolated spinal cord mitochondria: involvement of kinase-mediated pathways.

Ginsenoside Rd (Rd), one of the main active ingredients in Panax ginseng, has multifunctional activity via different mechanisms and neuroprotective effects that are exerted probably via its antioxidant or free radical scavenger action. However, the effects of Rd on spinal cord mitochondrial dysfunction and underlying mechanisms are still obscure. In this study, we sought to investigate the in vitro effects of Rd on mitochondrial integrity and redox balance in isolated spinal cord mitochondria. We verified that Ca2+ dissipated the membrane potential, provoked mitochondrial swelling and decreased NAD(P)H matrix content, which were all attenuated by Rd pretreatment in a dose-dependent manner. In contrast, Rd was not able to inhibit Ca2+ induced mitochondrial hydrogen peroxide generation. The results of Western blot showed that Rd significantly increased the expression of p-Akt and p-ERK, but had no effects on phosphorylation of PKC and p38. In addition, Rd treatment significantly attenuated Ca2+ induced cytochrome c release, which was partly reversed by antagonists of Akt and ERK, but not p-38 inhibitor. The effects of bisindolylmaleimide, a PKC inhibitor, on Rd-induced inhibition of cytochrome c release seem to be at the level of its own detrimental activity on mitochondrial function. Furthermore, we also found that pretreatment with Rd in vivo (10 and 50 mg/kg) protected spinal cord mitochondria against Ca2+ induced mitochondrial membrane potential dissipation and cytochrome c release. It is concluded that Rd regulate mitochondrial permeability transition pore formation and cytochrome c release through protein kinases dependent mechanism involving activation of intramitochondrial Akt and ERK pathways.

Detection of HLA-B*27 gene using a spectral plasmon resonance imaging system.

Detection of HLA-B*27 gene is clinically important due to its strong association with diseases, such as ankylosing spondylitis. Nucleic acid-based biosensors represent a promising clinical tool for gene diagnosis. Surface plasmon resonance (SPR) can be used to monitor DNA-DNA hybridization in real-time and without any prior labeling step. Here, a self-built HLA-B*27 positive PCR products spectral SPR imaging system was used for multichannel direct-detection of a specific sequence of the HLA-B*27 gene. Thiol-labeled single-stranded oligonucleotide probes, which were proved to be superior to amine-labeled probes in immobilization, were immobilized onto the surfaces of the gold spots on the sensor chip to target the specific sequence in the HLA-B*27 gene in blood. SPR measurements were performed with different concentrations of synthetic target DNA sequence. The calibration curve of synthetic target sequence showed a good linear relationship between the concentration of the synthetic target sequence and the shift of the SPR wavelength from 10 nM to 500 nM with a detection limit of 5 nM. The HLA-B*27 gene was isolated from human whole blood and amplified using polymerase chain reaction (PCR). PCR products were measured using the SPR imaging system. HLA-B*27 positive PCR products showed significant SPR wavelength shift, while synthetic non-complementary sequence and negative PCR products showed no significant SPR wavelength shift. The method is high-specific, high-throughput and label-free.

Detection of gastric carcinoma-associated antigen MG7-Ag in human sera using surface plasmon resonance sensor.

BACKGROUND: MG7-Ag is a kind of gastric cancer-specific tumor-associated antigen and has been investigated to serve as a marker of gastric cancer for early diagnosis. METHODS: Surface plasmon resonance (SPR) sensor was used for the detection of MG7-Ag in the sera of gastric cancer patients to develop an innovative, simple and rapid assay method for early diagnosis. The specific monoclonal MG7 antibodies were used as capture and detection receptors which were immobilized on the surface of SPR sensor chips for MG7-Ag identification in the human sera. The measurements include 9 cases of gastric cancer patients and 2 cases of healthy blood donors and a MKN45 cancer cell lysate solution sample for positive control. RESULTS: The binding of MG7-Ag onto the sensor surface was observed from SPR spectra. The sera of most gastric cancer patients revealed much higher expression level of MG7-Ag than healthy human sera did in SPR measurement. CONCLUSION: The initial results demonstrate that the SPR biosensor has the potential for its application in the early diagnosis of gastric cancer. However, more tests need to be done to confirm the detection limitation and the criterion for cancer risk evaluation in early diagnosis.