ObjectiveTo establish an allele-specific polymerase chain reaction （PCR） method for identifying Scolopendra dispensing granules， so as to ensure the quality and therapeutic effects of Scolopendra and its preparations. MethodThe primer interval suitable for the PCR was selected based on the cytochrome c oxidase subunit 3（COX-3） gene sequence of Scolopendra， and the single nucleotide polymorphism （SNP） loci of Scolopendra and its adulterants were mined from the interval for the design of specific primers. The samples of Scolopendra and its adulterants were collected. The PCR system was established and optimized regarding the annealing temperature， cycles， Taq enzymes， DNA template amount， PCR instruments， and primer concentrations， and the specificity and applicability of this method were evaluated. ResultThe PCR system was composed of 12.5 μL 2×M5 PCR Mix， 0.4 μL forward primer （10 μmol·L^-1）， 0.4 μL reverse primer （10 μmol·L^-1）， 2.5 μL DNA template， and 9.2 μL sterile double distilled water. PCR parameters： Pre-denaturation at 94 ℃ for 3 min， 30 cycles （94 ℃ for 20 s， 62 ℃ for 20 s， 72 ℃ for 45 s）， and extension at 72 ℃ for 5 min. After PCR amplification with the system and parameters above， the electrophoresis revealed a bright band at about 135 bp for Scolopendra and no band for the adulterants. ConclusionThe established allele-specific PCR method can accurately identify the medicinal materials， decoction pieces， and standard decoction freeze-dried powder of Scolopendra， as well as the intermediates and final products of Scolopendra dispensing granules， which is of great significance for ensuring the quality and clinical efficacy of Scolopendra and its preparations.

Objective To screen out specific aldosterone(ALD)antibodies using phage display technology and recom-binant antibody technology,providing raw materials for the research and development of ALD diagnostic kits.Methods Five healthy and clean New Zealand white rabbits were selected and immunized for the first time against the diluted ALD-keyhole limpet hemocyanin antigen(2 mg·L-1)using a multi-point injection method on the back,with a dose of 1 mg per rabbit.Immunization was administered again every 2 weeks,with a 50％reduction in dose.Starting from the third immunization,the ear vein blood of the rabbits was collected one week after each immunization.A chemiluminescent plate coated with 0.25 mg·L-1 ALD-bovine serum albumin antigen was used to measure serum titers via indirect and competitive methods.After the 5th immunization,the rabbit with high serum titers and good specificity was selected,and its spleen and bone marrow were removed.The spleen tissue was grinded,and RNA was extracted using TRIzol reagent in one step to obtain gene sequences in the variable region of light chain(VL)and the variable region of heavy chain(VH).The single-chain variable fragment(ScFv)was connected through the linker and constructed into the bacteriophage vector Pcomb3xss;then,it was carried to Escherichia coli TG1 through electrotransformation,and the ALD ScFv phage display library was constructed accordingly.Three to five rounds of enrichment screening were performed against the library.Monoclonal clones,identified by enzyme-linked immunosorbent assay(ELISA)competitive method,were selected for phage supernatant preparation,and a highly competitive clone sequence was obtained.The screened clone sequence was inserted into the pCMV3 expression vector,and the HEK293 cell was transfected using the transient transfection method after the plasmid was extracted.One week later,the supernatant was collected,and its purity and expression were identified by affinity chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE).Results After the 5th immunization,the serum titers of 5 rabbits were indirectly tested,and the results showed that the serum titers of 4# and 5# white rabbits were still greater than 10,000 after being diluted by 32,000 times.The test results based on the competitive method showed that the ratio of low to high values in the plasma sample of 5#white rabbit was 2∶1,superior to that of other white rabbits.The 5# white rabbit was selected for phage library construction.The VL and VH gene fragments were amplified by conventional polymerase chain reaction,and then bridged into ScFv(VL+VH).The agar gel electrophoresis analysis showed that the size of the band was about 750 bp,which was consistent with the size of the originally designed fragment.ScFv was cleaved and electroporated into Escherichia coli TG1 to construct a phage library with a storage capacity of 4.73 × 108 cfu·mL-1.After 3 rounds of washing,300 monoclonal clones were selected from the outbound petri dishes to prepare monoclonal bacteriophages.The ELISA results showed a positive rate of 100％among the 300 clones,and 42 clones were tested positive for calibration competition,with a screening rate of 14％.The 42 positive clones were further subjected to clinical sample competition testing,and 16 monoclonal strains that met the requirements were screened.The 16 strains were retested,and the results of the two tests were consistent.After sequencing,6 antibody sequences were selected for construction and expression.After purification,SDS-PAGE reduced gel electrophoresis results showed that there were bands at positions 50,000 on the heavy chain and 25,000 on the light chain.Six highly affinitive and competitive rabbit ALD monoclonal antibodies were obtained.Conclusion Six highly affinitive and competitive rabbit ALD antibodies are successfully screened using phage display technology,which provides a reference method for the discovery of small molecule antibodies.The screened AD1 85 and AD277 antibodies show a competitive advantage twice that of the positive control in the competition of calibration and clinical samples,providing a possibility for the development of raw materials for ALD detection kits.

Objective:To investigate the needle placement accuracy assisted by our self-designed channel screw guide system for pelvic fractures.Methods:Sixty pelvic models (provided by Shandong Weigao Group) were randomized into 2 groups ( n=30). In the experimental group, 2.0 mm Kirkler needles were implanted into the sacroiliac channel using our self-developed channel needle guide system in corresponding simulated surgical procedures; in the control group, the guide needles for sacroiliac screws were implanted manually under the guidance of C-arm fluoroscopy. The needle positions were assessed by gross observation and the offset distances measured on the X-ray films of pelvic entrance and exit views between the guide needle at the midline of the sacrum and the center point of bone channel. The offset distance, operation time, fluoroscopy frequency, and needle adjustment frequency were compared between the 2 groups of guide needles. Results:Guide needles were successfully implanted in all the pelvic models in the experimental group, with no penetration of guide needles outside the model. In the control group, 3 guide needles penetrated outside the model channel. The X-ray measurements showed that the offset distance of the needle in the experimental group was (2.23±0.82) mm, significantly smaller than that in the control group [(4.46±2.28) mm] ( P<0.05). In the experimental group, the fluoroscopy frequency [(12.0±0.3) times] and the needle adjustment frequency [(8.0±0.3) times] were significantly less than those in the control group [(26.0±0.4) times and (24.0±0.8) times] ( P<0.05). The operation time was (0.52±0.25) hours in the experimental group, significantly shorter than that in the control group [(1.26±0.36) hours] ( P<0.05). With a 2 mm diameter as an acceptable range, the accuracy was as high as 95.5%. Conclusion:Compared with manual placement of guide needles, our self-designed pelvic fracture channel screw guide system can lead to more accurate needle placement, reduced fluoroscopy frequency, fewer guide needle adjustments, and shortened operation time.

At present, most surgeons use release and subcutaneous transposition of the ulnar nerve to treat cubital tunnel syndrome (CuTS). However, subcutaneous transposition of the ulnar nerve is not effective enough and leads to a high rate of recurrence in clinic when CuTS is complicated with elbow osteoarthritis (EOA). Therefore, an appropriate surgical treatment should be adopted as early as possible if the symptoms are not relieved by a conservative treatment in cases of EOA complicated with CuTS. Different surgical methods lead to different physiological environments of the ulnar nerve, resulting in different outcomes. As a result, the recurrence rate of subcutaneous transposition motion is higher in patients with EOA. Surgical submuscular transposition of the ulnar nerve under the muscle significantly reduces its angular formation during elbow flexion, provides it with a more favorable biological environment and a better anatomical position, benefits the vascular reconstruction and regeneration of the injured nerve, and reduces its chance of secondary entrapment, leading to a better prognosis and a lower recurrence rate. This literature review evaluates the surgical methods for patients with CuTS plus EOA in an attempt to provide helpful references for their treatment.

Objective    To establish the gene-based esophageal cancer (ESCA) risk score prediction models via whole transcriptome analysis to provide ideas and basis for improving ESCA treatment strategies and patient prognosis. Methods    RNA sequencing data of esophageal squamous cell carcinoma (ESCC), esophageal adenocarcinoma (EAC) and adjacent tissues were obtained from The Cancer Genome Atlas database. The edgeR method was used to screen out the differential genes between ESCA tissue and normal tissue, and the key genes affecting the survival status of ESCC and EAC patients were initially identified through univariate Cox regression analysis. The least absolute shrinkage and selection operator regression analysis and multivariate Cox regression analysis were used to further screen genes and establish ESCC and EAC risk score prediction models. Results    The risk score prediction models were the independent prognostic factors for ESCA, and the risk score was significantly related to the survival status of patients. In ESCC, the risk score was related to T stage. In EAC, the risk score was related to lymph node metastasis, distant metastasis and clinical stage. The constructed nomogram based on risk score showed good predictive ability. In ESCC, the risk score was related to tumor immune cell infiltration and the expression of immune checkpoint genes. However, this feature was not obvious in EAC. Conclusion 聽 聽The ESCC and EAC risk score prediction models have shown good predictive capabilities, which provide certain inspiration and basis for optimizing the management of ESCA and improving the prognosis of patients.

Using chemoproteomic techniques, we first identified EIF2AK2, eEF1A1, PRDX3 and VPS4B as direct targets of berberine (BBR) for its synergistically anti-inflammatory effects. Of them, BBR has the strongest affinity with EIF2AK2 via two ionic bonds, and regulates several key inflammatory pathways through EIF2AK2, indicating the dominant role of EIF2AK2. Also, BBR could subtly inhibit the dimerization of EIF2AK2, rather than its enzyme activity, to selectively modulate its downstream pathways including JNK, NF-κB, AKT and NLRP3, with an advantage of good safety profile. In EIF2AK2 gene knockdown mice, the inhibitory IL-1β, IL-6, IL-18 and TNF-α secretion of BBR was obviously attenuated, confirming an EIF2AK2-dependent anti-inflammatory efficacy. The results highlight the BBR's network mechanism on anti-inflammatory effects in which EIF2AK2 is a key target, and inhibition of EIF2AK2 dimerization has a potential to be a therapeutic strategy against inflammation-related disorders.

Among ankle injuries, the injury to the distal tibiofibular syndesmosis is common and likely neglected. The stability of the distal tibiofibular syndesmosis is related to the depth of the fibular notch. In imaging diagnosis, X-ray examination cannot be used for a definite diagnosis of the injury to the distal tibiofibular syndesmosis. For diagnosis of the distal tibiofibular separation>3 mm, CT scan can be accurate but is not sensitive enough for a separation<1 mm while MRI is more sensitive in diagnosis of the injury. Arthroscopy has gradually been used as the "gold standard" in diagnosis of the injury to the distal tibiofibular syndesmosis due to its advantage of direct vision. The distal tibiofibular separation occurs in the injuries of pronation external rotation Ⅳ°, supination external rotation Ⅲ° and Ⅳ°, and pronation abduction Ⅱ° and Ⅲ° by the Lange-Hansen classification. Most patients with simple stable injury to the distal tibiofibular syndesmosis may have a good prognosis after nonoperative treatment. Surgical anatomic reduction and maintenance of stability of the distal tibiofibular syndesmosis are the basic management principles for unstable distal tibiofibular syndesmosis or the injury to the distal tibiofibular syndesmosis combined with ankle fracture. Screw fixation is the most commonly used in the surgical treatment of the injury. Elastic fixation has the advantages of maintaining the biological characteristics of the distal tibiofibular syndesmosis fretting joint, better reduction fault tolerance, and lower rates of complications and long-term reduction loss. The angle of nail placement is the key to maintaining good reduction of the distal tibiofibular syndesmosis, but there has been little description of the specific methods to ensure the theoretical angle of nail placement in practice. This article also reviews the prospects of the future treatment of the injury to the distal tibiofibular syndesmosis.

Objective:To understand the current situation of nurses' willingness to engage in telecare in county-level hospitals in Henan Province and analyze its influencing factors.Methods:Using the convenient sampling method, a total of 1 300 clinical nurses from 6 Class Ⅱ Grade A county-level hospitals in Zhengzhou, Hebi, Zhoukou and other cities in Henan Province were selected as the research objects from August 2022 to January 2023. General Data Questionnaire and Clinical Nurses' Willingness to Engage in Telecare Questionnaire were used to investigate the research objects. Non-parametric test and multiple linear regression were used to analyze the influencing factors of clinical nurses' willingness to engage in telecare.Results:A total of 1 300 questionnaires were sent out in this study, and 1 183 were effectively collected, with an effective recovery rate of 91.00% (1 183/1 300). The total score of Clinical Nurses' Willingness to Engage in Telecare Questionnaire of 1 183 nurses in county-level hospitals was [105.00 (98.00, 112.00) ] points, and all items were scored [3.50 (3.27, 3.73) ]. Multiple linear regression analysis showed that professional title, post, whether they knew about telecare, and the number of times of receiving training on nursing information each year were the influencing factors of nurses' willingness to engage in telecare in county-level hospitals ( P<0.05) . Conclusions:In Henan Province, 1 183 nurses in county-level hospitals have a high willingness to engage in telecare. Relevant departments should further strengthen the publicity of relevant contents of telecare, carry out practical operation training related to nursing information, improve the nursing information ability of nurses in county-level hospitals and strengthen the awareness of health management services, so as to promote the successful implementation of telecare services in rural areas.

ObjectiveTo observe the behavioral and pain threshold alterations， as well as the changes in indexes related to depression and pain in the serum and central system in mice stressed by maternal separation and chronic neuropathic pain， and explore the underlying mechanism of Wenyang prescription （WY）， Jieyu prescription （JY）， and Wenyang Jieyu prescription （WYJY） in improving depression and pain sensitivity. MethodThe birth date of mice was recorded as PD0. After birth， the mice were divided into a blank group and an experimental group. The neonatal mice in the experimental group underwent maternal separation in PD5-14 at 8 h·d^-1. After ablactation， the mice were divided into a maternal separation group， a WY group （Erxian decoction， 5.84 g·kg^-1）， a JY group （Xiaoyaosan， 12.00 g·kg^-1）， a WYJY group （16.68 g·kg^-1）， and a fluoxetine group （2.60 mg·kg^-1）， with 15 mice in each group. Meanwhile， 15 male mice of the same age without maternal separation were assigned to the normal control group. Mice in the blank group and the maternal separation group were fed on a regular chow diet in PD21-PD90， while the remaining groups were fed on the corresponding drugs. In PD91， sciatic nerve ligation was performed to induce a model of maternal separation and chronic neuropathic pain. The open field test was used to observe the depression-like behaviors of mice in each group， and the mechanical and temperature pain thresholds were measured to detect the pain sensitivity of mice in each group. The serum levels of corticosterone （CORT）， substance P， and β-endorphin （β-EP） were determined by enzyme-linked immunosorbent assay （ELISA）， and the expression of the glucocorticoid receptor （GR） in the amygdala and β-EP protein in the hypothalamus was detected by immunohistochemistry. The mRNA expression levels of amygdala GR gene （Nr3c1）， FK506 binding protein 5 gene （FKBP5）， metabolic glutamate receptor 5 gene （GRM5）， and brain-derived neurotrophic factor （BDNF） were detected by real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）. ResultCompared with the blank group， the maternal separation group showed reduced stay time and total distance traveled in the 5-min open field test （P<0.01）， reduced mechanical pain threshold （P<0.01）， increased serum CORT and β-EP （P<0.01）， declining FKBP5 mRNA expression （P<0.01）， and increased hypothalamic β-EP expression （P<0.05）. Compared with the maternal separation group， the groups with drug intervention showed prolonged stay time （P<0.05， P<0.01） and up-regulated pain thresholds to different degrees. The total distance traveled in the 5-min open field test increased in the WY group， the WYJY group， and the fluoxetine group （P<0.05， P<0.01）. The JY group showed decreased serum CORT （P<0.01）， reduced β-EP ， and increased BDNF mRNA （P<0.01）. Nr3c1 and GRM5 mRNA decreased in the WY group （P<0.05， P<0.01）. The WYJY group showed decreased serum CORT （P<0.05）and decreased Nr3c1， GRM5， and BDNF mRNA （P<0.05， P<0.01）. The levels of β-EP expression were elevated to different degrees in the groups with drug intervention， but the differences were not significant. The levels of GR expression in the WY group， the JY group， and the WYJY group increased （P<0.05）. ConclusionWYJY can inhibit central pain sensitization and regulate hypothalamic-pituitary-adrenal gland （HPA） axis function by enhancing the expression of GR in the amygdala and inhibiting neuroplasticity and excitability in the amygdala to relieve depression-like behaviors and improve somatic hyperalgesia.

Objective:To investigate the current situation of moisture-associated skin damage (MASD) in patients with permanent colostomy, and analyze its influencing factors.Methods:From January 2019 to January 2020, 115 patients received permanent colostomy in Henan Provincial People's Hospital were selected by convenience sampling method as the research objects. The patients were investigated by the self-designed General Information Questionnaire and Disease Factor Questionnaire.Results:Finally, 107 patients with permanent colostomy were included, and the incidence of MASD was 39.25% (42/107) . One-way analysis of variance showed that age, body mass index, stoma site, stoma height, excrement characteristics, defecation regularity, chassis leakage times were the influencing factors for the incidence of MASD in patients with permanent colostomy ( P<0.05) . Conclusions:The incidence of MASD in patients with permanent colostomy is high. Age, stoma site, stoma height, defecation and so on are the influencing factors of MASD in patients with permanent colostomy. Medical and nursing staff can formulate targeted preventive measures according to the influencing factors to reduce the incidence of MASD.