CRISPR/Cas-mediated "one to more" lighting-up nucleic acid detection using aggregation-induced emission luminogens.

CRISPR diagnostics are effective but suffer from low signal transduction efficiency, limited sensitivity, and poor stability due to their reliance on the trans-cleavage of single-stranded nucleic acid fluorescent reporters. Here, we present CrisprAIE, which integrates CRISPR/Cas reactions with "one to more" aggregation-induced emission luminogen (AIEgen) lighting-up fluorescence generated by the trans-cleavage of Cas proteins to AIEgen-incorporated double-stranded DNA labeled with single-stranded nucleic acid linkers and Black Hole Quencher groups at both ends (Q-dsDNA/AIEgens-Q). CrisprAIE demonstrates superior performance in the clinical nucleic acid detection of norovirus and SARS-CoV-2 regardless of amplification. Moreover, the diagnostic potential of CrisprAIE is further enhanced by integrating it with spherical nucleic acid-modified AIEgens (SNA/AIEgens) and a portable cellphone-based readout device. The improved CrisprAIE system, utilizing Q-dsDNA/AIEgen-Q and SNA/AIEgen reporters, exhibits approximately 80- and 270-fold improvements in sensitivity, respectively, compared to conventional CRISPR-based diagnostics. We believe CrisprAIE can be readily extended as a universal signal generation strategy to significantly enhance the detection efficiency of almost all existing CRISPR-based diagnostics.

Recovery quality of transversus abdominis plane block with liposomal bupivacaine after cesarean delivery: A randomized trial.

STUDY OBJECTIVE: This study aimed to investigate the effect of liposomal bupivacaine in transversus abdominis plane block (TAP) on recovery quality after cesarean delivery. DESIGN: A randomized trial. SETTING: An operating room, a post-anesthesia care unit, and a hospital ward. PATIENTS: A total of 147 women scheduled for cesarean delivery under spinal anesthesia were enrolled and randomized to receive a TAP block with plain bupivacaine (bupivacaine group), liposomal bupivacaine (liposomal group), or a mixture of plain bupivacaine and liposomal bupivacaine (mixture group). INTERVENTIONS: The bupivacaine group received bilateral TAP blocks with plain bupivacaine 50 mg alone. The liposomal group received bilateral TAP blocks with liposomal bupivacaine 266 mg alone. The mixture group received bilateral TAP blocks with plain bupivacaine 50 mg followed by liposomal bupivacaine 266 mg. MEASUREMENTS: The primary outcome was the Quality of Recovery-15 (QoR - 15) score assessed 24 h postoperatively. Secondary outcomes encompassed the QoR - 15 score at 48 h post-surgery, the VAS pain score at rest and with movement at 24, 48, and 72 h postoperatively, opioid consumption within the 0-24 h and 24-48 h periods following surgery, as well as patient's satisfaction with analgesic. MAIN RESULTS: The QoR - 15 score at 24 h postoperatively was significantly higher in both the liposomal group and the mixture group compared to the bupivacaine group. Specifically, the QoR - 15 score for the liposomal group versus the bupivacaine group (median [IQR]: 120 [107, 128] vs. 109 [104, 120]; median difference, 7; 95 % CI, 2 to 13; P = 0.011) and for the mixture group versus the bupivacaine group (median [IQR]: 122 [112, 128] vs. 109 [104, 120]; median difference, 9; 95 % CI, 4 to 14; P = 0.001). The QoR - 15 score in both the liposomal group and the mixture group were also higher than those in the bupivacaine group at 48 h postoperatively, though the difference was not clinically meaningful. Additionally, both the liposomal and mixture groups exhibited lower pain score at 24 h and 48 h postoperatively compared to the bupivacaine group, but no significant clinical differences were achieved in either pain scores or opioid consumption. Patients in both the liposomal and mixture groups reported higher satisfaction score with analgesia than those in the bupivacaine group. CONCLUSIONS: TAP block using either liposomal bupivacaine or a mixture of plain bupivacaine and liposomal bupivacaine provided superior quality of recovery at 24 h after cesarean delivery compared to using plain bupivacaine alone.

Effect of optimizing cerebral oxygen saturation on postoperative delirium in older patients undergoing one-lung ventilation for thoracoscopic surgery.

OBJECTIVES: This randomized controlled trial investigated whether the regional cerebral oxygenation saturation (rScO2)-guided lung-protective ventilation strategy could improve brain oxygen and reduce the incidence of postoperative delirium (POD) in patients older than 65 years. METHODS: This randomized controlled trial enrolled 120 patients undergoing thoracic surgery who received one-lung ventilation (OLV). Patients were randomly assigned to the lung-protective ventilation group (PV group) or rScO2-oriented lung-protective ventilation group (TPV group). rScO2 was recorded during the surgery, and the occurrence of POD was assessed. RESULTS: The incidence of POD 3 days after surgery-the primary outcome-was significantly lower in the TPV group (23.3% versus 8.5%). Meanwhile, the levels of POD-related biological indicators (S100ß, neuron-specific enolase, tumor necrosis factor-α) were lower in the TPV group. Considering the secondary outcomes, both groups exhibited a lower oxygenation index after OLV, whereas partial pressure of carbon dioxide and mean arterial pressure were significantly increased in the TPV group. In addition, minimum rScO2 during surgery and mean rScO2 were higher in the TPV group than in the PV group. CONCLUSION: Continuous intraoperative monitoring of brain tissue oxygenation and active intervention measures guided by cerebral oxygen saturation are critical for improving brain metabolism and reducing the risk of POD.

Pulley Effect in the Capture of DNA Translocation through Solid-State Nanopores.

Nanopores are powerful single-molecule sensors for analyzing biomolecules such as DNA and proteins. Understanding the dynamics of DNA capture and translocation through nanopores is essential for optimizing their performance. In this study, we examine the effects of applied voltage and pore diameter on current blockage, translocation time, collision, and capture location by translocating λ-DNA through 5.7 and 16 nm solid-state nanopores. Ionic current changes are used to infer DNA conformations during translocation. We find that translocation time increases with pore diameter, which can be attributed to the decrease of the stall force. Linear and exponential decreases of collision frequency with voltage are observed in the 16 and 5.7 nm pores, respectively, indicating a free energy barrier in the small pore. Moreover, the results reveal a voltage-dependent bias in the capture location toward the DNA ends, which is explained by a "pulley effect" deforming the DNA as it approaches the pore. This study provides insights into the physics governing DNA capture and translocation, which can be useful for promoting single-file translocation to enhance nanopore sensing.

Establishment of a hydrodynamic delivery system in ducks.

Chronic hepatitis B virus (HBV) poses a significant global health challenge as it can lead to acute or chronic liver disease and hepatocellular carcinoma (HCC). To establish a safety experimental model, a homolog of HBV-duck HBV (DHBV) is often used for HBV research. Hydrodynamic-based gene delivery (HGD) is an efficient method to introduce exogenous genes into the liver, making it suitable for basic research. In this study, a duck HGD system was first constructed by injecting the reporter plasmid pLIVE-SEAP via the ankle vein. The highest expression of SEAP occurred when ducks were injected with 5 µg/mL plasmid pLIVE-SEAP in 10% bodyweight volume of physiological saline for 6 s. To verify the distribution and expression of exogenous genes in multiple tissues, the relative level of foreign gene DNA and ß-galactosidase staining of LacZ were evaluated, which showed the plasmids and their products were located mainly in the liver. Additionally, ß-galactosidase staining and fluorescence imaging indicated the delivered exogenous genes could be expressed in a short time. Further, the application of the duck HGD model on DHBV treatment was investigated by transferring representative anti-HBV genes IFNα and IFNγ into DHBV-infected ducks. Delivery of plasmids expressing IFNα and IFNγ inhibited DHBV infection and we established a novel efficient HGD method in ducks, which could be useful for drug screening of new genes, mRNAs and proteins for anti-HBV treatment.

Metagenomic next-generation sequencing for the identification of infections caused by Gram-negative pathogens and the prediction of antimicrobial resistance.

OBJECTIVE: The aim of this study was to evaluate the efficacy of metagenomic next-generation sequencing (mNGS) for the identification of Gram-negative bacteria (GNB) infections and the prediction of antimicrobial resistance. METHODS: A retrospective analysis was conducted on 182 patients with diagnosis of GNB infections who underwent mNGS and conventional microbiological tests (CMTs). RESULTS: The detection rate of mNGS was 96.15%, higher than CMTs (45.05%) with a significant difference (χ 2 = 114.46, P < .01). The pathogen spectrum identified by mNGS was significantly wider than CMTs. Interestingly, the detection rate of mNGS was substantially higher than that of CMTs (70.33% vs 23.08%, P < .01) in patients with but not without antibiotic exposure. There was a significant positive correlation between mapped reads and pro-inflammatory cytokines (interleukin-6 and interleukin-8). However, mNGS failed to predict antimicrobial resistance in 5 of 12 patients compared to phenotype antimicrobial susceptibility testing results. CONCLUSIONS: Metagenomic next-generation sequencing has a higher detection rate, a wider pathogen spectrum, and is less affected by prior antibiotic exposure than CMTs in identifying Gram-negative pathogens. The mapped reads may reflect a pro-inflammatory state in GNB-infected patients. Inferring actual resistance phenotypes from metagenomic data remains a great challenge.

An integrated mRNA-lncRNA signature for overall survival prediction in cholangiocarcinoma.

The combination of mRNA and lncRNA profiles for establishing an integrated mRNA-lncRNA prognostic signature has remained unexplored in cholangiocarcinoma (CCA) patients. We utilized a training dataset of 36 samples from The Cancer Genome Atlas dataset and a validation cohort (GSE107943) of 30 samples from Gene Expression Omnibus. Two mRNAs (CFHR3 and PIWIL4) and 2 lncRNAs (AC007285.1 and AC134682.1) were identified to construct the integrated signature through a univariate Cox regression (P-value = 1.35E-02) and a multivariable Cox analysis (P-value = 3.07E-02). Kaplan-Meier curve showed that patients with low risk scores had notably prolonged overall survival than those with high risk scores (P-value = 4.61E-03). Subsequently, the signature was validated in GSE107943 cohort with an area under the curve of 0.750 at 1-year and 0.729 at 3-year. The signature was not only independent from diverse clinical features (P-value = 3.07E-02), but also surpassed other clinical characteristics as prognostic biomarkers with area under the curve of 0.781 at 3-year. Moreover, the weighted gene co-expression network analysis and gene enrichment analyses found that the integrated signature were associated with metabolic-related biological process and lipid metabolism pathway, which has been implicated in the pathogenesis of CCA. Taken together, we developed an integrated mRNA-lncRNA signature that had an independent prognostic value in the risk stratification of patients with CCA.

Association between PD-L1 and Ki-67 expression and clinicopathologic features in NSCLC patients.

OBJECTIVE: To investigate the relationship between PD-L1, Ki-67 and the association between PD-L1, Ki-67, and clinicopathologic features and laboratory parameters in the peripheral blood of non-small cell lung cancer (NSCLC) patients. METHODS: The clinical records of 213 NSCLC patients were retrospectively reviewed. The patients were divided into high or low expression groups by cut-off values of Ki-67 and PD-L1. Correlation of PD-L1 and Ki-67 expression were analyzed by linear regression analysis. The data were tested by Mann-Whitney test. Relationship of PD-L1 and Ki-67 was tested by chi-square test. RESULTS: Linear regression analysis revealed a positive association between the expression of PD-L1 and Ki-67 (R2=0.26, P<0.001). The clinicopathologic features and laboratory parameters such as gender, smoking history, histological types, TNM stage, tumor size, ALB, and FIB were all significantly associated with PD-L1 and Ki-67 expression (all P<0.05). CONCLUSIONS: The expression of PD-L1 and Ki-67 is related to some clinicopathologic features and inflammatory factors, which brings new sight for exploiting combination biomarkers and therapeutic strategies.

Application of the oblique lateral interbody fusion technique in salvage surgery: technical note and case series.

Objective: The oblique lateral interbody fusion (OLIF) technique is a promising interbody fusion technique. This study summarizes the technical aspects of OLIF as a salvage surgery and the preliminary outcomes of a series of cases. Patients and methods: A retrospective review of patients with leg or back pain induced by pseudoarthrosis or adjacent segment disease after posterior lumbar interbody fusion/transforaminal lumbar interbody fusion was done. These patients underwent salvage OLIF surgeries in our institution from January 2021 to March 2022. Variables such as the demographic, clinical, surgical, and radiological characteristics of the enrolled patients were recorded and analyzed. Results: Eight patients (five females and three males; mean age 69.1 ± 5.7 years, range 63-80 years) were enrolled in this study. The mean operative time was 286.25 min (range: 230-440 min), and the estimated blood loss was 90 ml (range: 50-150 ml). Only one of the eight patients experienced a complication of lower limb motor weakness, which disappeared within 5 days after surgery. The latest data showed that the mean intervertebral space height increased from 8.36 mm preoperatively to 12.70 mm and the mean segmentary lordosis increased from 8.92° preoperatively to 15.05°. Bone fusion was achieved in all but one patient, who was followed up for only 3 months. The JOA scores Japanese Orthopaedic Association (JOA) Scores for low back pain of all patients significantly improved at the final follow-up. Conclusion: OLIF provides a safe and effective salvage strategy for patients with failed posterior intervertebral fusion surgery. Patients effectively recovered intervertebral and foraminal height with no additional posterior direct decompression.

Clinical Efficacy and Diagnostic Value of Metagenomic Next-Generation Sequencing for Pathogen Detection in Patients with Suspected Infectious Diseases: A Retrospective Study from a Large Tertiary Hospital.

Purpose: Metagenomic next-generation sequencing (mNGS) is a powerful yet unbiased method to identify pathogens in suspected infections. However, little is known about its clinical effectiveness. The present study aimed to assess the efficacy of mNGS in routine clinical practice. Patients and Methods: In this single-center retrospective cohort study, 518 patients with suspected infectious diseases were assessed for inclusion. Among them, each patient had undergone mNGS testing; 407 patients had undergone both microbial culture and mNGS testing. The result of mNGS testing was compared to microbial culture performed concurrently. The diagnostic performance of mNGS was evaluated using the comprehensive clinical diagnosis as the reference standard. Results: There was a significant difference in the positive detection rates of pathogens between mNGS and culture (331/407, 81.3% vs 79/407, 19.4%, P < 0.001). The sensitivity of mNGS was much higher than the culture method (79.5% vs 21.3%, P < 0.001), especially in sample types of sputum and bronchoalveolar lavage fluid (BALF). Notably, the sensitivity of blood mNGS was relatively lower than other sample types (67.4% vs 88.9-93.8%). Pathogen cfDNA load based on standardized stringently mapped read number at the species level of microorganisms (SDSMRN) was significantly lower in blood than in other sample types from the same patient (P = 0.0003). Importantly, mNGS directly led to a change of treatment regimen in 142 (27.4%) cases, including antibiotic escalation (15.3%), antibiotic de-escalation (9.1%), and early definitive diagnosis to initiate appropriate treatment (3.1%). Conclusion: Our in-house mNGS platform significantly improved the sensitivity for the diagnosis of infectious diseases. mNGS has the potential to improve clinical outcomes by optimizing antimicrobial therapy.

Comparison of Lumbar Fusion Surgical Outcomes Between Patients with Lumbar Spinal Stenosis ≥80 Versus 65-79 Years Old.

BACKGROUND The efficacy of lumbar fusion surgery in patients age 80 years and older with lumbar spinal stenosis (LSS) is still controversial. This retrospective study aimed to evaluate the surgical outcomes of LSS patients ³80 vs 65-79 years. MATERIAL AND METHODS The study included 66 patients diagnosed with LSS from 2014 to 2020; 33 patients were ³80 years and 33 patients were 65-79 years. The 2 groups were matched for sex and surgical segment. All patients underwent posterior lumbar decompression, fixation, and fusion surgery. The Numerical Rating Scale (NRS) of leg and back pain, Oswestry Disability Index (ODI), Short-Form Health Survey (SF-36), and radiographic data were collected before surgery and at 3, 6, and 12 months postoperatively. RESULTS NRS (back) in the ≥80 years group was significantly higher than in the 65-79 years group at 3 months [2 (0-3) vs 1 (0-3), P=0.001]. Improvement of SF-36 (3 months: 15.7±4.9 vs 27.6±5.4, P<0.001; 6 months: 27.3±6.8 vs 31.5±5.6, P=0.011) and Physical Component Score (PCS) (3 months: 6.5±2.5 vs 17.0±3.6, P<0.001; 6 months: 15.9±3.4 vs 20.1±3.1, P<0.001) at 3 and 6 months in the ≥80 years group were significantly smaller than in 65-79 years group. There was a difference of Pfirrmann index of adjacent segment disc between the 2 groups at 12 months [≥80 vs 65-79 years group: 5.5 (4-8) vs 5 (3-8), P=0.003]. CONCLUSIONS Lumbar fusion surgery in patients ≥80 years with LSS can provide comparable improvements in clinical and radiographic outcomes compared with younger patients. Postoperative physiological function recovery was slower in patients ≥80 years.

A rare discal cyst with concurrent double-level isthmic lumbar spondylolisthesis: a case report and literature review.

We presented a 47-year old male diagnosed as a discal cyst with concurrent isthmic lumbar spondylolisthesis, which was not reported in the literature previously. A cystectomy with two-segmental transforaminal lumbar interbody fusion and instruments was performed. The association between concurrent discal cyst and isthmic spondylolisthesis may be illuminated by excessive strain and focal degeneration of the disc. Preoperative discography is probably unnecessary when surgical resection and histopathology are indicated for the patient.

Surgical Precision and Efficiency of a Novel Electromagnetic System Compared to a Robot-Assisted System in Percutaneous Pedicle Screw Placement of Endo-LIF.

STUDY DESIGN: Retrospective database study. OBJECTIVES: To compare the accuracy and safety of 2 types of a computer-assisted navigation system for percutaneous pedicle screw placement during endoscopic lumbar interbody fusion. METHODS: From May 2019 to January 2020, data of 56 patients who underwent Endo-LIF with a robot-assisted system and with an electromagnetic navigation system were compared. The pedicles in all patients were subjected to postoperative CT scan to assess screw correction by measuring the perpendicular distance between the pedicle cortical wall and the screw surface. The registration and matching time, guide-wire insertion time, the entire surgery time, and X-ray exposure time were recorded. RESULTS: In the robot-assisted group, 25 cases with 100 percutaneous pedicle screws were included, and the excellent and good rate was 95%. In the electromagnetic navigation group, 31 cases with 124 screws were included, and the excellent rate was 97.6%. There was no statistical difference between the two groups (P > 0.05). The registration time and the total time for the surgery also showed no statistical differences (P > 0.05). The main difference between the two groups was the guide-wire insertion time and the X-ray exposure time (P < 0.05). CONCLUSIONS: Both electromagnetic navigation and robot-assisted are safe and efficient for percutaneous pedicle screw placement. Electromagnetic navigation system has obvious advantages over robot-assisted in terms of faster guide-wire placement and less X-ray exposure. Robot-assisted for percutaneous pedicle screw placement offers a preoperative planning system and a stable registration system, with obvious drawbacks of a strict training curve.

Fully Endoscopic 360° Decompression for Central Lumbar Spinal Stenosis Combined with Disc Herniation: Technical Note and Preliminary Outcomes of 39 Cases.

Purpose: To evaluate the outcomes, feasibility, and safety of endoscopic unilateral laminectomy, bilateral decompression and discectomy (Endo-ULBDD) for central lumbar spinal stenosis (CLSS) combined with disc herniation (DH). Methods: This study includes 39 patients diagnosed with CLSS combined with DH who met the inclusion criteria and underwent surgery for Endo-ULBDD from April 2020 to March 2021. The mean age of the patients, operation time, hospitalization time, time in bed, and complications were recorded. Patients were followed up for at least 12 months. Visual analog scale (VAS) scores for low-back and lower-limb pain and Oswestry Disability Index (ODI) scores were evaluated preoperatively, before discharge, and at 3, 6, and 12 months postoperatively. To evaluate clinical effectiveness 12 months postoperatively, the modified MacNab criteria were used. Results: The mean age of the patients was 59.9 years, the mean operation time was 82.1 minutes, the mean hospitalization time was 3.7 days, and the mean time in bed was 20.9 hours. The mean VAS scores of low-back and lower-limb pain improved from 5.9 and 7.2 to 2.0 and 1.6, respectively (P < 0.05). The ODI score improved from 56.0 to 16.7 (P < 0.05). The overall excellent-good rate of the modified MacNab criteria was 89.7%. Two kinds of complications occurred in 4 patients (10.3%), including 1 patient whose inferior articular process was excessively removed and 3 patients who suffered from postoperative dysesthesia. No other severe complications were noted. Conclusion: Endo-ULBDD is a safe, feasible, efficient, and minimally invasive approach to treating CLSS combined with DH.

Application of electromagnetic navigation in endoscopic transforaminal lumbar interbody fusion: a cohort study.

STUDY DESIGN: Clinical retrospective cohort study. OBJECTIVES: To explore the application of the electromagnetic navigation system in Endo-TLIF. MATERIALS AND METHODS: From May 2019 to March 2020, 76 patients with single-segment lumbar spondylolisthesis treated by electromagnetic navigation-assisted Endo-TLIF (NE group) and conventional Endo-TLIF (CE group) were enrolled in the study. Time of pedicle screw implantation, entire operation time, the number of intraoperative X-ray fluoroscopy exposures, total blood loss, incision length, ambulation time, accuracy of pedicle screws, complications, visual analog scale for back and leg pain, Oswestry Disability Index, Japanese Orthopedic Association score and postoperative fusion rates were recorded, respectively. RESULTS: There were no significant differences in preoperative demographics between the NE and CE groups (P > 0.05). The mean number of intraoperative X-ray fluoroscopy exposures, guidewires insertion, entire operation time, total blood loss and adjustment rate of screws in the NE group were significantly less compared with the CE group (P < 0.05, respectively). There were no significant differences in clinical parameters between the two groups at different time points in the follow-up period (P > 0.05). There was no statistical difference in fusion rates between the two groups. In addition, one case of cage subsidence was observed after surgery in the CE group. CONCLUSION: Electromagnetic navigation systems could be applied throughout the entire surgical course and ameliorate the shortcomings of the conventional Endo-TLIF technique to reduce radiation exposure, improve accuracy, avoid repetitive operations and shorten surgery time and the required learning curve of the procedure. LEVEL OF EVIDENCE I: Diagnostic: individual cross-sectional studies with consistently applied reference standard and blinding.

Comparison of electromagnetic and optical navigation assisted Endo-TLIF in the treatment of lumbar spondylolisthesis.

Uniportal full endoscopic posterolateral transforaminal lumbar interbody fusion (Endo-TLIF) with percutaneous pedicle screw fixation is a promising, minimally invasive method for the treatment of lumbar spondylolisthesis. However, repeated radiation exposure from X-rays and the steep learning curve remain to be improved.

Expression of Potato StDRO1 in Arabidopsis Alters Root Architecture and Drought Tolerance.

Potato (Solanum tuberosum L) is the third important crop for providing calories to a large human population, and is considered sensitive to moderately sensitive to drought stress conditions. The development of drought-tolerant, elite varieties of potato is a challenging task, which can be achieved through molecular breeding. Recently, the DEEPER ROOTING 1 (DRO1) gene has been identified in rice, which influences plant root system and regulates grain yield under drought stress conditions. The potato StDRO1 protein is mainly localized in the plasma membrane of tobacco leaf cells, and overexpression analysis of StDRO1 in Arabidopsis resulted in an increased lateral root number, but decreased lateral root angle, lateral branch angle, and silique angle. Additionally, the drought treatment analysis indicated that StDRO1 regulated drought tolerance and rescued the defective root architecture and drought-tolerant phenotypes of Atdro1, an Arabidopsis AtDRO1 null mutant. Furthermore, StDRO1 expression was significantly higher in the drought-tolerant potato cultivar "Unica" compared to the drought-sensitive cultivar "Atlantic." The transcriptional response of StDRO1 under drought stress occurred significantly earlier in Unica than in Atlantic. Collectively, the outcome of the present investigation elucidated the role of DRO1 function in the alternation of root architecture, which potentially acts as a key gene in the development of a drought stress-tolerant cultivar. Furthermore, these findings will provide the theoretical basis for molecular breeding of drought-tolerant potato cultivars for the farming community.

Identification and Expression Analysis of Calcium-Dependent Protein Kinases Gene Family in Potato Under Drought Stress.

Calcium-dependent protein kinases (CDPKs) are a class of serine/threonine protein kinases encoded by several gene families that play key roles in stress response and plant growth and development. In this study, the BLAST method was used to search for protein sequences of the potato Calcium-dependent protein kinase gene family. The chromosome location, phylogeny, gene structures, gene duplication, cis-acting elements, protein-protein interaction, and expression profiles were analyzed. Twenty-five CDPK genes in the potato genome were identified based on RNA-seq data and were clustered into four groups (I-IV) based on their structural features and phylogenetic analysis. The result showed the composition of the promoter region of the StCDPKs gene, including light-responsive elements such as Box4, hormone-responsive elements such as ABRE, and stress-responsive elements such as MBS. Four pairs of segmental duplications were found in StCDPKs genes and the Ka/Ks ratios were below 1, indicating a purifying selection of the genes. The protein-protein interaction network revealed defense-related proteins such as; respiratory burst oxidase homologs (RBOHs) interacting with potato CDPKs. Transcript abundance was measured via RT-PCR between the two cultivars and their relative expression of CDPK genes was analyzed after 15, 20, and 25 days of drought. There were varied expression patterns of StCDPK3/13/21 and 23, between the two potato cultivars under mannitol induced-drought conditions. Correlation analysis showed that StCDPK21/22 and StCDPK3 may be the major differentially expressed genes involved in the regulation of malondialdehyde (MDA) and proline content in response to drought stress, opening a new research direction for genetic improvement of drought resistance in potato.

Relationship between hinge fracture and postoperative axial symptoms after cervical laminoplasty.

OBJECTIVE: To identify the clinical characteristics of fractured hinges after open-door cervical laminoplasty for cervical canal stenosis and explore the relationship between hinge fractures and axial symptoms. METHODS: This was a retrospective study of patients with cervical myelopathy who underwent open-door laminoplasty between November 2014 and November 2016 at the Affiliated Hospital of Qingdao University. Cervical CT scans were performed after surgery and the Takeuchi criteria were applied to evaluate the postoperative axial symptoms. RESULTS: Of 223 opened laminae in 67 patients, 67 laminae (30.0%) in 30 patients (44.8%) showed fracture. The frequency of hinge fractures was higher at C6 (53.7%). Forty-nine fractured laminae (73.13%) were non-displaced and 18 were displaced. At 3 months, 33 fractured laminae (49.3%) showed bony union on CT, and union rates were 86.6% and 91.0% at 6 and 12 months, respectively, indicating that the union rate was lower for displaced fractures than for non-displaced fractures. Among the 67 patients, 14 had axial symptoms: three of 37 (8.1%) patients without hinge fractures and 11 of 30 (36.7%) patients with hinge fractures. One year later, the hinge fractures were healed in 24/30 patients. Among the six unhealed patients, five still suffered from axial symptoms. The frequency of axial symptoms was higher in the patients with three or more hinge fractures (66.7%) than in the patients with only one (16.7%) or two (46.7%) hinge fractures. CONCLUSIONS: Patients with hinge fractures may have an increased risk for axial symptoms after open-door cervical laminoplasty. The frequency of axial symptoms decreases with fracture healing.