Objective:To evaluate the sample preparation proficiency and storage proficiency of 11 clinical biobanks in Beijing through simulated experiments, and to establish an assessment method for the quality comparability of biological samples.Methods:An exploratory research design was adopted. In November 2023, artificial composite serum quality control materials containing six recombinant human protein markers—recombinant human alanine aminotransferase (rhALT), recombinant human aspartate aminotransferase (rhAST), recombinant human creatine kinase (rhCK), recombinant human creatine kinase-MB (rhCK-MB), recombinant human B-type natriuretic peptide (rhBNP), and recombinant human troponin I (rhTNI)—were distributed to 11 clinical biobanks in Beijing City. Sample preparation and storage followed the standardized operating procedures. Proficiency differences were assessed through statistical analysis.Results:Three-way repeated measures ANOVA revealed all six protein markers showed a declining trend over storage time in ultra-low-temperature environments ( F values 11.68-4 179.66, all P<0.01). However, neither long-term/temporary refrigerator types ( F values 0.01-1.23, all P>0.05)nor placement locations within refrigerators significantly affected the stability of these six proteins ( F valus 0.03-1.47, all P>0.05). The biases in detection results for rhALT, rhAST, rhTNI, and rhBNP at different storage time points were within the allowable bias limits for each item, supporting their use as markers for protein stability in biobank samples. All 11 institutions passed the storage proficiency assessment. In the preparation proficiency assessment, deviations were observed in post-preparation sample results, with a notably high out-of-control rate for rhCK (36.36%). Conclusion:Sample preparation proficiency can serve as a quality control metric for clinical biobanks. Future external quality assessment systems for biobanks should focus on sample preparation rather than storage processes.

Objective:To analyze the APR triangle formed when the right hepatic vein (RHV) passed through the right anterior hepatic pedicle (RAHP) and the right posterior hepatic pedicle (RPHP) by 3D visualization technique.Methods:Clinical data of 100 subjects undergoing liver contrast-enhanced CT scan in the Second Affiliated Hospital of Harbin Medical University from September 2019 to September 2022 were retrospectively analyzed, including 57 males and 43 females, aged (53.2±13.0) years. The CT data were imported into a system for 3D image reconstruction of the liver. The intrahepatic vascular structure was observed, including the portal vein, APR triangle, the angle between the RAHP and RPHP, the distance between RHV and the vertex of angle when it crosses the angle plane of the RAHP and RPHP, the structure and branches of the right posterior branch of portal vein, and the number of branches into the APR triangle from the main trunk of the right anterior and right posterior portal vein.Results:The main type of portal vein was type A (the main portal vein was divided into left and right branches, and the right branch was further divided into right anterior branch and right posterior branch), in 86 cases (86.0%). The APR triangle was observed in 81 cases (81.0%), and the distance between the right hepatic vein and the vertex of the right anterior and right posterior hepatic pedicle was (1.70±0.36) cm, 1.00-2.00 cm was 82.7%(61/81). The angle between right anterior and right posterior hepatic pedicle was 45°-90° in 77 cases (77.0%, 77/100). In 100 subjects, the main branch of the right posterior portal vein accounted for 42.0% (42/100) of typeⅠ (bifurcation type) and 53.0% (arch type) of type Ⅱ (53/100). In 16 cases (19.8%, 16/81), the main right anterior portal vein branched into the APR triangle, with a total of 22 branches. In 9 cases (11.1%, 9/81), the main branch of the right posterior portal vein (branching type) branched into the triangle, with a total of 11 branches. There were 33 cases (40.7%, 33/81) of right posterior portal vein (arch type) main branch branching into the triangle, a total of 41 branches.Conclusion:The APR triangle, composed of the RAHP, RPHP and RHV, has an objective anatomical basis and is affected by the variants of portal vein and RHV. It is more common for the RHV to be within 1.00-2.00 cm of the vertex of the angle between the RAHP and RPHP, and the main trunk of the right anterior and right posterior branches of portal vein are less divided into the triangle, so it is relatively safe to dissect within this area.

Objective:To evaluate the sample preparation proficiency and storage proficiency of 11 clinical biobanks in Beijing through simulated experiments, and to establish an assessment method for the quality comparability of biological samples.Methods:An exploratory research design was adopted. In November 2023, artificial composite serum quality control materials containing six recombinant human protein markers—recombinant human alanine aminotransferase (rhALT), recombinant human aspartate aminotransferase (rhAST), recombinant human creatine kinase (rhCK), recombinant human creatine kinase-MB (rhCK-MB), recombinant human B-type natriuretic peptide (rhBNP), and recombinant human troponin I (rhTNI)—were distributed to 11 clinical biobanks in Beijing City. Sample preparation and storage followed the standardized operating procedures. Proficiency differences were assessed through statistical analysis.Results:Three-way repeated measures ANOVA revealed all six protein markers showed a declining trend over storage time in ultra-low-temperature environments ( F values 11.68-4 179.66, all P<0.01). However, neither long-term/temporary refrigerator types ( F values 0.01-1.23, all P>0.05)nor placement locations within refrigerators significantly affected the stability of these six proteins ( F valus 0.03-1.47, all P>0.05). The biases in detection results for rhALT, rhAST, rhTNI, and rhBNP at different storage time points were within the allowable bias limits for each item, supporting their use as markers for protein stability in biobank samples. All 11 institutions passed the storage proficiency assessment. In the preparation proficiency assessment, deviations were observed in post-preparation sample results, with a notably high out-of-control rate for rhCK (36.36%). Conclusion:Sample preparation proficiency can serve as a quality control metric for clinical biobanks. Future external quality assessment systems for biobanks should focus on sample preparation rather than storage processes.

Objective:To analyze the APR triangle formed when the right hepatic vein (RHV) passed through the right anterior hepatic pedicle (RAHP) and the right posterior hepatic pedicle (RPHP) by 3D visualization technique.Methods:Clinical data of 100 subjects undergoing liver contrast-enhanced CT scan in the Second Affiliated Hospital of Harbin Medical University from September 2019 to September 2022 were retrospectively analyzed, including 57 males and 43 females, aged (53.2±13.0) years. The CT data were imported into a system for 3D image reconstruction of the liver. The intrahepatic vascular structure was observed, including the portal vein, APR triangle, the angle between the RAHP and RPHP, the distance between RHV and the vertex of angle when it crosses the angle plane of the RAHP and RPHP, the structure and branches of the right posterior branch of portal vein, and the number of branches into the APR triangle from the main trunk of the right anterior and right posterior portal vein.Results:The main type of portal vein was type A (the main portal vein was divided into left and right branches, and the right branch was further divided into right anterior branch and right posterior branch), in 86 cases (86.0%). The APR triangle was observed in 81 cases (81.0%), and the distance between the right hepatic vein and the vertex of the right anterior and right posterior hepatic pedicle was (1.70±0.36) cm, 1.00-2.00 cm was 82.7%(61/81). The angle between right anterior and right posterior hepatic pedicle was 45°-90° in 77 cases (77.0%, 77/100). In 100 subjects, the main branch of the right posterior portal vein accounted for 42.0% (42/100) of typeⅠ (bifurcation type) and 53.0% (arch type) of type Ⅱ (53/100). In 16 cases (19.8%, 16/81), the main right anterior portal vein branched into the APR triangle, with a total of 22 branches. In 9 cases (11.1%, 9/81), the main branch of the right posterior portal vein (branching type) branched into the triangle, with a total of 11 branches. There were 33 cases (40.7%, 33/81) of right posterior portal vein (arch type) main branch branching into the triangle, a total of 41 branches.Conclusion:The APR triangle, composed of the RAHP, RPHP and RHV, has an objective anatomical basis and is affected by the variants of portal vein and RHV. It is more common for the RHV to be within 1.00-2.00 cm of the vertex of the angle between the RAHP and RPHP, and the main trunk of the right anterior and right posterior branches of portal vein are less divided into the triangle, so it is relatively safe to dissect within this area.

Objective To explore the effect of open reading frame 66(C12ORF66)located at chromosome 12 on the viability of MYCN amplified NB cell lines.Methods DDatasets GSE16476 and GSE49710 in R2 database were analyzed for expression level of C12ORF66 in MYCN amplified and MYCN non-amplified NB cells and its potential correlation with the prognosis of pediatric patients.C12ORF66 mRNA expression level in normal tissue immortalized cell lines,MYCN amplified and MYCN non-amplified cell lines were detected by RT-qRCR.Transient or stable knockdown of C12ORF66 cell lines were constructed to compare the difference in real time cellular analysis(RTCA),colony formation,Ki67 positive cells between the control group and the C12ORF66 knockdown group.Results By analyzing R2 datasets,C12ORF66 level in MYCN amplified samples was significantly higher than that in MYCN non-amplified samples,and the expression of C12ORF66 was negatively correlated with the prognosis of pediatric patients(P＜0.05).C12ORF66 highly expressed in MYCN-amplified BE(2)-C and SK-N-BE(2)cell lines than in MYCN non-amplified CHLA-255 and SH-SY5Y cell lines(P＜0.001).Transient or stable knockdown of C12ORF66 resulted in significant slow down of proliferation of MYCN amplified NB cells(P＜0.001),the colony formation ability was significantly reduced(P＜0.001),and the proportion of Ki67 positive cells was significantly decreased(P＜0.05).Conclusions C12ORF66 was highly expressed in MYCN amplified clinical NB samples and cell lines which is believed to be correlated with poor prognosis of pediatric patients.C12ORF66 knockdown signifi-cantly inhibits cell viability of NB cells.

The cell cycle is a complex process that involves DNA replication, protein expression, and cell division. Dysregulation of the cell cycle is associated with various diseases. Cyclin-dependent kinases (CDKs) and their corresponding cyclins are major proteins that regulate the cell cycle. In contrast to inhibition, a new approach called proteolysis-targeting chimeras (PROTACs) and molecular glues can eliminate both enzymatic and scaffold functions of CDKs and cyclins, achieving targeted degradation. The field of PROTACs and molecular glues has developed rapidly in recent years. In this article, we aim to summarize the latest developments of CDKs and cyclin protein degraders. The selectivity, application, validation and the current state of each CDK degrader will be overviewed. Additionally, possible methods are discussed for the development of degraders for CDK members that still lack them. Overall, this article provides a comprehensive summary of the latest advancements in CDK and cyclin protein degraders, which will be helpful for researchers working on this topic.
Humans ; Cell Cycle/physiology* ; Cell Division ; Cyclin-Dependent Kinases/metabolism* ; Cyclins/metabolism*

Objective:To evaluate the effect of sleep deprivation on cognitive function in septic rats and its relationship with neuronal glycolysis isoenzyme phosphofructokinase-2/fructose-2,?6-diphosphatase 3 (PFKFB3).Methods:Fifty-six healthy male Sprague-Dawley (SD) rats were randomly divided into 4 groups ( n = 14): control group (Con group), sepsis group (LPS group), sepsis+sleep deprivation group (LPS+SD group), sepsis+sleep deprivation+glycolysis inhibitor 3-PO treatment group (LPS+SD+3-PO group). The sepsis model was established by intraperitoneal injection of lipopolysaccharide (LPS) 10 mg/kg. Rats in LPS+SD group were treated with sleep deprivation using a sleep deprivation instrument 24 hours after LPS injection. The LPS+SD+3-PO group was intraperitoneally injected with LPS for 24 hours, and then injected with 3-PO 50 mg/kg, followed by sleep deprivation. Novel object recognition experiments were performed 72 hours after LPS injection. Subsequently, blood and brain tissue samples were collected. The contents of lactate (Lac), reactive oxygen species (ROS) and serum tumor necrosis factor-α(TNF-α), neuron-specific enolase (NSE), pyruvate in brain tissue were detected by enzyme-linked immunosorbent assay (ELISA). Then, the lactate/pyruvate ratio was calculated. Na +-K +-ATPase activity in brain tissue was detected by colorimetry. Morphological changes in hippocampus were detected by hematoxylin-eosin (HE) staining. And the protein expression levels of PFKFB3, ZO-1 and cleaved caspase-3 were measured by Western blotting. Results:Compared with Con group, the novel object recognition index of LPS group was decreased, the levels of NSE, TNF-α, lactate/pyruvate ratio in serum and the levels of Lac, ROS and dry-wet weight ratio in brain tissue were significantly increased, Na +-K +-ATPase activity in brain tissue was decreased, the protein expressions of PFKFB3, caspase-3 were up-regulated, ZO-1 expression was down-regulated, and the neurons in hippocampus were slightly degenerated. Compared with LPS group, the novel object recognition index of LPS+SD group was further decreased [(39.4±5.3)% vs. (54.5±7.6)%)], serum NSE, TNF-α, lactate/pyruvate ratio and brain tissue Lac, ROS, dry-wet weight ratio were further increased [NSE (μg/L): 3.21±0.42 vs. 2.55±0.36, TNF-α (ng/L): 139.4±19.7 vs. 92.2±13.5, lactate/pyruvate ratio: 29.7±5.5 vs. 19.2±4.2, Lac (μmol/g): 19.51±2.33 vs. 11.34±1.52, ROS (kU/g): 117.4±18.7 vs. 78.2±11.8, dry-wet weight ratio: (81.3±9.2)% vs. (64.3±6.6)%], and Na +-K +-ATPase activity was further decreased (mmol·L -1·h -1: 1.88±0.34 vs. 2.91±0.39), the protein expressions of PFKFB3, caspase-3 were further up-regulated and ZO-1 expression was further down-regulated (PFKFB3/β-actin: 0.80±0.11 vs. 0.45±0.07, caspase-3/β-actin: 0.71±0.09 vs. 0.37±0.05, ZO-1/β-actin: 0.31±0.05 vs. 0.61±0.08). The differences were statistically significant (all P < 0.05). HE staining showed that the degeneration of neurons in hippocampus was significantly aggravated. Compared with LPS+SD group, the novel object recognition index of LPS+SD+3-PO group was increased [(50.8±5.9)% vs. (39.4±5.3)%], NSE, TNF-α, lactate/pyruvate ratio of serum and Lac, ROS, dry-wet weight ratio of brain tissue were significantly decreased [NSE (μg/L): 2.60±0.33 vs. 3.21±0.42, TNF-α (ng/L): 103.7±18.3 vs. 139.4±19.7, lactate/pyruvate ratio: 17.4±5.1 vs. 29.7±5.5, Lac (μmol/g): 13.68±2.02 vs. 19.51±2.33, ROS (kU/g): 86.9±14.5 vs. 117.4±18.7, dry-wet weight ratio: (67.7±6.9)% vs. (81.3±9.2)%], and Na +-K +-ATPase activity was increased (mmol·L -1·h -1: 2.82±0.44 vs. 1.88±0.34). The protein expressions of PFKFB3, caspase-3 were down-regulated and ZO-1 expression was up-regulated (PFKFB3/β-actin: 0.50±0.06 vs. 0.80±0.11, caspase-3/β-actin: 0.43±0.06 vs. 0.71±0.09, ZO-1/β-actin: 0.52±0.06 vs. 0.31±0.05). The differences were statistically significant (all P < 0.05). HE staining showed that the degeneration of neurons in hippocampus was significantly improved. Conclusions:Sleep deprivation could aggravate neuroinflammation, neuronal degeneration and apoptosis in septic rats, resulting in destruction of blood-brain barrier and cognitive impairment. 3-PO treatment significantly alleviate the injury and degeneration of hippocampal neurons in septic rats, inhibit neuroinflammation and apoptosis, and improve cognitive dysfunction, which may be related to the inhibition of glycolytic isoenzyme PFKFB3.

Objective:To elucidate the spatial distribution patterns of the right hepatic vein by analyzing the image information obtained after CT three-dimension reconstruction of liver to provide guidance in surgical planning of anatomical hepatectomy.Methods:A retrospective analysis was performed on the clinical data of 77 subjects who underwent CT examination of the liver at the Second Affiliated Hospital of Harbin Medical University from September 2018 to October 2021. There were 42 males and 35 females, aged (50.2±12.8) years old. CT DICOM data of the patients were collected, and the two-dimensional image data were reconstructed into a three-dimensional model by using the 3D reconstruction software. The characteristics and typing were studied by analyzing the number of branches of the right hepatic vein and the spatial location of the main trunk.Results:Of 77 subjects, 645 branches of the right hepatic vein were observed in the liver CT 3D reconstruction model, including 268 (41.6%) right-sided branches, 240 (37.2%) dorsal branches, 70 (10.9%) left-sided branches, and 67 (10.3%) ventral branches. Each right hepatic vein possessed 3 (3, 4) right-sided branches, 3 (3, 4) dorsal branches, 1 (0, 1) left-sided branch, and 1 (0, 1) ventral branch. The numbers of branches in the four directions were significantly different ( H=175.89, P<0.001). Comparison showed that the number of right-sided branches was significantly more than that of the left-sided (χ 2=136.86) and ventral (χ 2=140.07), respectively. The number of dorsal branches was more than that of left-sided (χ 2=-123.36) and ventral (χ 2=126.57) branches, respectively. The differences were significant ( P<0.001). There were no significant differences between the number of ventral and left-sided branches, and between the dorsal and right-sided branches (all P>0.05). Conclusion:The right hepatic vein had fewer ventral and left-sided branches. It is relatively safe to dissect the right hepatic vein from the ventral or the left side during surgery. For resection of the central liver segments or segment VIII of the liver, it is reasonable to transect the liver along the left border of the right hepatic vein.

ObjectiveTo identify the functions of the AP2/ERF family members in Pinellia ternata and promote the genetic improvement of P. ternata varieties. MethodWe identified and conducted a systematic bioinformatics analysis of the AP2/ERF family member genes in P. ternata based on the three generations of transcriptome data. Real-time polymerase Chain reaction　（Real-time） PCR was employed to determine the expression pattern of AP2/ERF genes in different tissues and under different stress conditions. ResultA total of eight full-length AP2/ERF family members were identified from the transcriptome data， which were classified into three sub-gene families： AP2， ERF， and DREB. The deduced AP2/ERF proteins in P. ternata had the length of 251-512 aa， the theoretical pI of 5.29-11.72， the instability index of 45.90-82.41， subcellular localization in the nucleus， and conserved domains and motifs. AP2/ERF genes were expressed in different tissues of P. ternata， with high expression levels in the leaf. The stress response experiments showed that PtERF1 mainly responded to NaCl stress. The expression of PtERF2 and PtERF4 was significantly up-regulated under low temperature and polyethylene glycol （PEG）-simulated stress. PtERF3 responded to both low temperature and NaCl stress. The expression of PtERF5 was induced by high temperature， low temperature， NaCl and PEG stress. The expression of PtERF7 was up-regulated under high temperature， while that of PtERF8 under low temperature. ConclusionThe AP2/ERF genes in P. ternata can respond to stress and have the potential functions of regulating photosynthesis and improving root stress resistance.

Objective:To explore the feasibility and safety of long-distance urological nephrotomy with the support of 5G communication technology by using the domestic robot.Methods:Clinical data of the patients with remote robot-assisted laparoscopic nephrectomy, which were completed from March to April 2021 by the Affiliated Hospital of Qingdao University (as the host hospital where the main operating system located) were retrospectively analyzed. There were 3 patients, including 2 males and 1 female.The average age was 61 (49-73) years, and the average body mass index was 23.73 (20.00-27.76) kg/m 2. One patient had a ASA classification of grade 2, and the other 2 patients had grade 3. All patients met the surgical criteria for non-functional nephrectomy. The chief surgeon who performing the telesurgery was located at the Affiliated Hospital of Qingdao University. The surgeon remotely controlled the bedside operating system (slave system) in 3 local hospitals located in other cities in Shandong Province (network communication distances of 82.5, 141 and 229 km, respectively) by manipulating the master system located in Qingdao. Images and operating instructions during surgery were transmitted using 5G wireless communication technology. Intraoperative network conditions, robot operation, and patient perioperative data were summarized. Results:All 3 tele-nephrectomies were successfully completed. The average network signal latency time was 27.3 (23-30) ms, with no packet loss, and the average total latency time was 177.3(173-180) ms. The mean resection time was 79.3 (52-111) min, and the average intraoperative blood loss was 31.1 (15.6-41.9) ml. There were no network related adverse events occurred during the operation, and the robot-related adverse events occured 3 times, all three of which were characterized by inconsistent master and slave movements of the manipulator arm and the bedside robotic arm. None of these adverse events affected the successful performance of the telesurgery. The mean postoperative exhaust time was 60.5 (38.5-78.0) h. The mean postoperative VAS score at 24 hours was 3.7 (3-4). The Clavien-Dindo classification were all grade I. No significant abnormality was found on the 30th day after surgery, and the patients recovered well at the follow-up until 6 months postoperatively.Conclusions:It is safe and feasible to perform remote robot-assisted laparoscopic nephrectomy based on 5G communication technology with no serious adverse events or surgical complications.However, the conclusion needs to be further verified by large sample and multi-center prospective study.