ObjectiveTo investigate the role and mechanism of Go-Ichi-Ni-San complex subunit 1 （GINS1） in the progression of hepatocellular carcinoma （HCC） and the development of chemotherapy resistance. MethodsThe tumor database GEPIA2 was used to analyze the differential expression of GINS1 between HCC patients and healthy individuals， and pathological tissue samples were collected from 40 HCC patients who were admitted to The Affiliated Tumor Hospital of Xinjiang Medical University and the First Affiliated Hospital of Xinjiang Medical University from May 2017 to January 2021. Immunohistochemical staining was used to measure the difference in the expression of GINS1 between HCC tissue and corresponding adjacent tissue， and the correlation between the expression level of GINS1 and the clinical TNM stage of HCC was analyzed. Western blot was also used to measure the difference in the expression of GINS1 between HCC Huh7/Hep3B/Li-7/MHCC97H cell lines and normal human QSG7701 hepatocytes. The method of lentivirus transfection was used to establish the MHCC97H cell line with stable GINS1 knockdown and its negative control cell line. CCK-8 assay and colony formation assay were used to measure cell proliferative capacity； scratch assay was used to measure cell migration ability； Transwell assay was used to measure cell invasion ability； cells were treated with oxaliplatin to measure their sensitivity to chemotherapy drugs. Nude mice were used to establish a tumor-bearing model and observe the effect of GINS1 knockdown on the growth of HCC in vivo. Western Blot was used to measure the expression levels of the proteins associated with the Notch pathway and the JAK/STAT pathway. The cells were treated with the Notch receptor agonist Jagged-1 to analyze the association between GINS1 and the Notch/JAK/STAT pathway. The independent-samples t test was used for comparison of continuous data between two groups； a one-way analysis of variance was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsThe expression of GINS1 was upregulated in HCC patients， HCC tissue， and HCC cell lines （all P<0.05）， and the expression level of GINS1 was positively correlated with the clinical TNM stage of HCC （r=0.822， P=0.011）. Compared with the negative control cells， the GINS1-knockdown MHCC97H cells showed significant reductions in proliferation， migration， and invasion activities （all P<0.01） and a significantly enhanced sensitivity to oxaliplatin （P<0.01）. Compared with the nude mice in the control group， GINS1 knockdown caused significant inhibition of tumor weight and volume in vivo in nude mice （all P<0.001）. Compared with the negative control cells， the GINS1-knockdown MHCC97H cells showed significant reductions in the expression levels of Notch1， Notch3， p-JAK2， and p-STAT3 （all P<0.05）， while there were no significant differences in the overall expression levels of JAK2 and STAT3 （P>0.05）. After Jagged-1 treatment， the GINS1-knockdown MHCC97H cells showed significant increases in proliferation， migration， and invasion activities and a significant reduction in sensitivity to oxaliplatin， as well as significant increases in the levels of p-JAK2 and p-STAT3 （all P<0.05）. ConclusionGINS1 is upregulated in HCC and can promote HCC progression and chemotherapy resistance through the Notch/JAK2/STAT3 pathway.

Objective:To investigate the relationship between the timing of pulmonary surgery and postoperative pulmonary complications (PPCs) after severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection.Methods:Sixty-eight American Society of Anesthesiologists Physical Status classification Ⅰor Ⅱ patients of either sex, with body mass index of 18-30 kg/m 2, who were first infected with SARS-CoV-2 after December 2022, undergoing elective thoracoscopic partial pneumonectomy from January to May 2023, were included in this prospective cohort study. The patients were divided into 2 groups ( n=34 each) according to the time between the date of surgery and SARS-CoV-2 infection: 5-10 weeks group and 11-16 weeks group. The preoperative persistent symptoms and dyspnea before operation were recorded. The serum concentrations of interleukin-6 and tumor necrosis factor-alpha were determined by enzyme-linked immunosorbent assay at 1 day before operation and 2 h and 1 and 2 days after operation. The white blood cell count and serum C-reactive protein concentration were measured at 1 day before operation and 1 and 2 days after operation. The occurrence of PPCs and length of postoperative hospital stay were recorded. Logistic regression was used to analyze the relationship between PPCs and timing of pulmonary surgery after SARS-CoV-2 infection. Results:Two patients in each group were excluded from the study because of conversion to thoracotomy. Thirty-two patients were finally included in each group. Compared with 5-10 weeks group, the ratio of preoperative persistent symptoms and dyspnea was significantly decreased, the serum concentrations of interleukin-6, tumor necrosis factor-alpha and C-reactive protein and white blood cell count were decreased at each time point after operation, the incidence of PPCs and postoperative pulmonary infection was decreased, and the length of postoperative hospital stay was shortened in 11-16 weeks group ( P<0.05). Multivariate logistic regression analysis showed that short time from the date of surgery to infection ( OR=1.754, 95% confidence interval[ CI] 1.509-2.038, P<0.001), preoperative persistent symptoms ( OR=2.523, 95% CI 2.047-3.110, P<0.001), preoperative dyspnea ( OR=1.875, 95% CI 1.406-2.500, P<0.001) and high white blood cell count at 1 day after surgery ( OR=0.676, 95% CI 0.651-0.701, P<0.001) were independent risk factors for PPCs. Conclusions:The risk of PPCs is lower in the patients undergoing pulmonary surgery at 11-16 weeks after SARS-CoV-2 infection than at 5-10 weeks after infection. Short time from the date of surgery to infection is an independent risk factor for PPCs.

Objective To observe the effect of pressure-controlled ventilation volume-guaranteed(PCV-VG)mode on respiratory mechanics,lung injury markers and postoperative pulmonary complications(PPCs)in thoracoscopic patients.Methods Fifty-nine patients undergoing elective thoracoscopic lobecto-my,29 males and 30 females,aged 18-64 years,BMI 18.5-26.0 kg/m2,ASA physical status Ⅰ or Ⅱ,were divided into two groups using a random number table method:the PCV-VG mode group(group P,n=29)and the volume-controlled ventilation(VCV)mode group(group V,n=30).The PCV-VG mode was used for one-lung ventilation(OLV)in group P,and the VCV mode was used in group V.Anesthesia in-duction and maintenance medications were consistent in all patients.PaO2 was recorded before induction of anesthesia,5 minutes after intubation,15 minutes after OLV,30 minutes after OLV,and 3 days postopera-tively,and oxygenation index(OI)and intrapulmonary shunt rate(Qs/Qt)were calculated.Peak airway pressure(Ppeak),pulmonary dynamic compliance(Cdyn),and driving pressure(DP)were recorded 5 minutes after intubation,15 minutes after OLV,and 30 minutes after OLV.Clara cell secretory protein-16(CC-16)and interleukin-6(IL-6)concentration were measured before induction of anesthesia and after ex-tubation.Recording the occurrence of PPCs within 1 week after surgery.Results Compared with group V,Ppeak and DP were significantly reduced,Cdyn was increased significantly in group P 15 minutes and 30 minutes after OLV(P＜0.05),PaO2 and OI were significantly increased in group P 3 days postoperatively(P＜0.05),CC-16 and IL-6 concentrations were significantly reduced in group P after extubation(P＜0.05).Compared with group V,the incidence of PPCs was significantly reduced in group P(P＜0.05).Conclusion During one-lung ventilation for thoracoscopic surgery,the pressure-controlled ventilation vol-ume-guaranteed mode reduces peak airway pressure and driving pressure,improves pulmonary dynamic compliance and improves oxygenation,reduces the incidence of PPCs.

Objective To investigate the correlation between different operation timing of thoraco-scopic partial pneumonectomy and intraoperative intrapulmonary shunt rate in patients with novel coronavirus(SARS-CoV-2)infection.Methods A total of 120 patients,65 males and 55 females,aged 30-75 years,BMI 18.5-25.0 kg/m2,ASA physical status Ⅰ or Ⅱ,scheduled for elective thoracoscopic partial pneumo-nectomy from December 2022 to May 2023 were selected.The patients with SARS-CoV-2 infection were di-vided into three groups according to different operation timing after infection:5-8 weeks after infection(group B),9-12 weeks after infection(group C),and 13-16 weeks after infection(group D),30 pa-tients in each group.In addition,30 non-infected patients were selected as the control group(group A).Blood gas analysis was performed at 10 minutes of two-lung ventilation(TLV)and 15 and 30 minutes of one-lung ventilation(OLV)to measure radial artery and mixed venous blood gases.Intrapulmonary shunt rate(Qs/Qt)was calculated accordingly.Multiple linear regression analysis was used to investigate the cor-relation between different operation timing and intrapulmonary shunt rate in patients with SARS-CoV-2 infec-tion.The occurrence of postoperative pulmonary complications(PPCs)within 7 days after surgery was re-corded.Results Compared with group A,groups B and C exhibited significant decreases in PaO2 levels and increases in Qs/Qt ratios at 10 minutes of TLV as well as at 15 and 30 minutes of OLV(P＜0.05),group D exhibited significant decreases in PaO2 levels and increases in Qs/Qt ratios at 15 and 30 minutes of OLV(P＜0.05),group B exhibited significant increases in postoperative pulmonary infection rates and the incidence of respiratory failure within 7 days after surgery(P＜0.05).Compared with group B,the inci-dence of pulmonary infection and respiratory failure within 7 days after surgery were significantly reduced in group D(P＜0.05).Multiple linear regression analysis revealed that shorter infection time(β=-0.478,95％CI-3.857 to-1.231,P＜0.001),worsening clinical types of infection(β=0.274,95％CI 0.368 to 3.453,P=0.016),and preoperative persistent symptoms(β=-0.240,95％CI-5.986 to-0.537,P=0.019)were associated with increased intrapulmonary shunt rate at 10 minutes of TLV.Shor-ter infection time(β=0.267,95％CI 0.130 to 3.018,P=0.033),worsening clinical types of infection(β=-0.391,95％CI-4.715 to-1.323,P=0.001),preoperative persistent symptoms(β=-0.497,95％CI-10.484 to-4.491,P＜0.001),and preoperative dyspnea(β=-0.246,95％CI-8.596 to-0.691,P=0.022)were associated with increased intrapulmonary shunt rate at 15 minutes of OLV.Conclusion SARS-CoV-2 infection increases intrapulmonary shunt rate 5-8 and 9-12 weeks after infection,but the intrapulmonary shunt rate gradually recovers at 10 minutes of TLV 13-16 weeks after in-fection,and patients who undergo surgery during this interval have a lower incidence of PPCs.The shorter infection time,the aggravation of clinical classification of infection,and the presence of persistent symptoms before surgery are associated with the increase of intrapulmonary shunt rate.

Objective To investigate the risk factors affecting the malignancy of amorphous calcifications in the breast and to establish a predictive nomogram.Methods Patients with amorphous calcifications detected by mammography were retrospectively collected,clinical data were obtained from electronic medical record(EMR),and the mammographic features of the patients were assessed by diagnostic physicians.The risk factors affecting the malignancy of amorphous calcifications were analyzed to develop a predictive model and to assess the performance of the model.Results A total of 153 amorphous calcifications in 144 patients were included in the study,and the overall malignancy rate of calcifications was 20.92％.Patient's age ≥45 years,linear distribution of calcifications,unilateral single or unilateral multiple calcifications,and a larger maximum ratio of calcification extent all predicted a higher probability of malignancy,establishing a nomogram based on these 4 risk factors,with a 3.65％predicted probability of malig-nancy as the cut-off,33.99％(52/153)of patients were allowed to be spared biopsy.Conclusion Patient's age and the distribution,number,and maximum ratio of calcifications may be the risk predictors of malignancy for amorphous calcifications,with nomogram con-struction for distinguishing benignity from malignancy of amorphous calcifications via combining with mammographic features and clinical data.

OBJECTIVE Notoginseng Radix et Rhizoma residues were used as raw material to compare the difference in the im-pact of different probiotics and composite probiotic on various indicator components through probiotic fermentation,aiming to explore the optimal fermentation process.METHODS The fermentation process was optimized using single factor and Box-Behnken response surface methodology,and the antioxidant capacity of the fermentation product was assessed through in vitro antioxidant experiments.RESULTS The results showed the optimum fermentation processes were 48 h of aerobic fermentation,36 h of anaerobic fermentation,ratio of 2∶3∶1 for Streptococcus thermophilus,Bifidobacterium bifidum and Lactobacillus acidophilus,solid-liquid ratio of 0.14 g·mL-1,inoculation quantity 5%,fermentation temperature 33℃.Under the optimal fermentation conditions,the content of neutral polysaccharide,acidic polysaccharide,and total flavonoid in Notoginseng Radix et Rhizoma residue increased by 105.64%,96.98%and 123.83%,respectively,which were high than those single-strain fermentation.The IC50 values of scavenging DPPH and ABTS free radicals in the fermentation products were 1.774 mg·mL-1 and 3.065 mg·mL-1,respectively,and the power of reducing Fe3+was 0.138 mmol FeSO4 g-1.The antioxidant capacity was significantly enhanced compared to the unfermented residues.CON-CLUSION The optimum fermentation process can significantly elevate the content of indicator components in Notoginseng Radix et Rhizoma residues and enhance its antioxidant capacity.Compared to single-strain fermentation,the content of various indicator compo-nents is significantly increased,showing no apparent antagonistic effect among the probiotics mentioned above.The study provides sci-entific evidence and data support for the resource utilization of Notoginseng Radix et Rhizoma residues.

Objective:To detect expression levels of complement regulatory proteins CD55 and CD59 in the peripheral blood of patients with Stevens-Johnson syndrome (SJS) /toxic epidermal necrolysis (TEN), and to preliminarily analyze their potential roles in the occurrence of SJS/TEN.Methods:Hospitalized patients with SJS/TEN (SJS/TEN group) were collected from the Department of Dermatology of the First Affiliated Hospital of Anhui Medical University and the Second Affiliated Hospital of Anhui Medical University from December 2017 to December 2022. Meanwhile, patients with maculopapular exanthema (MPE) and healthy physical examinees were also collected and served as the mild group and healthy control group, respectively. Flow cytometry was performed to determine the proportions of CD4 + T lymphocytes and CD8 + T lymphocytes in peripheral blood mononuclear cells (PBMCs). The expression levels of inflammatory cytokines tumor necrosis factor (TNF) -α, interleukin (IL) -6, IL-17, IL-10, interferon (IFN) -γ, IL-2, and IL-4 were detected using flow cytometric bead array technology. The mRNA expression levels of CD55 and CD59 in PBMCs were detected by real-time fluorescence-based quantitative PCR (qRT-PCR). Flow cytometry was also performed to determine the protein expression of CD55 and CD59 on the surface of CD8 + T lymphocytes. Statistical analyses were carried out using one-way analysis of variance and Tukey's test. Results:Totally, 13 patients with SJS/TEN, 27 patients with MPE, and 40 healthy controls were collected. Among the SJS/TEN patients, there were 8 males and 5 females, with their age being 18 to 84 (47.15 ± 19.99) years, and disease duration being 7.74 ± 2.63 days. No significant differences were observed in the gender distribution or age among the 3 groups (both P > 0.05). The proportions of CD4 + T lymphocytes did not differ among the 3 groups ( F = 3.84, P = 0.051). The proportions of CD8 + T lymphocytes in the peripheral blood were significantly higher in the SJS/TEN group (25.60% ± 4.57%) than in the healthy control group (16.20% ± 6.28%; q = 4.59, P = 0.018). The expression levels of inflammatory cytokines TNF-α, IL-6, IL-10, IL-17, and IFN-γ were significantly higher in the SJS/TEN group than in the healthy control group and mild group (all P < 0.001). In addition, the mRNA expression of CD55 ( F = 9.46, P < 0.001) and CD59 in PBMCs ( F = 15.14, P < 0.001) was significantly lower in the SJS/TEN group than in the mild group and healthy control group. The protein expression levels of CD55 ( F = 51.51, P < 0.001) and CD59 ( F = 31.59, P < 0.001) on the surface of CD8 + T lymphocytes were also significantly lower in the SJS/TEN group than in the other two groups and the healthy control group, respectively. Conclusion:Complement regulatory proteins CD55 and CD59 were downregulated in SJS/TEN patients, which may be associated with the activation of CD8 + T lymphocytes and excessive inflammatory responses.

Objective:To analyze the clinical characteristics and prognostic factors of high-risk neuroblastoma (HR-NB) patients with skeletal metastasis.Methods:The clinical features of 336 newly treated HR-NB patients with skeletal metastases admitted to the Department of Medical Oncology of Beijing Children′s Hospital, Capital Medical University from January 2007 to December 2018 were analyzed retrospectively.Kaplan-Meier method was used for the survival analysis, and Log- Rank test was used for univariate prognosis analysis.The Cox model was used to analyze the multifactorial prognostic analysis. Results:A total of 336 HR-NB patients were recruited, involving 188 males and 148 females with the median age of onset of at 43 (4-148) months.Skeletal metastases affected the viscerocranium (89 cases, 26.5%), neurocranium (193 cases, 57.4%), vertebrae (298 cases, 88.7%), sternum and ribs (183 cases, 54.5%), pelvis (270 cases, 80.4%), upper limbs (182 cases, 54.2%) and lower limbs (240 cases, 71.4%). The 5-year event-free survival (EFS) rate and overall survival (OS) rate were (30.4±2.7)% and (41.3±2.9)%, respectively.Univariate analysis showed a significantly lower 5-year OS rate in skeletal metastatic HR-NB patients with poor prognostic classification, the morphology of neuroblastoma (stroma-poor) and ganglioneuroblastoma (intermixed), high index of mitosis-karyorrhexis index, lactate dehydrogenase≥587 U/L, serum ferritin≥92 μg/L, MYCN amplification and 1p loss of heterozygosity, and metastases in the viscerocranium, neurocranium, vertebrae, sternum and ribs, pelvis, upper limbs and lower limbs (all P<0.05). The 5-year OS rate of HR-NB patients with all 7 regions of skeletal metastases was only (14.2±5.9)%, which was significantly lower than that in patients with a single region metastasis or multi-region metastases[(66.0±10.2)% vs.(43.6±3.4)%, χ2=45.722, P<0.05]. Cox multifactorial analysis showed that MYCN amplification ( HR=4.165, 95% CI: 2.356-7.363) and the viscerocranium metastasis ( HR=2.560, 95% CI: 1.519-4.315) were the independent risk factors affecting the prognosis of HR-NB patients with skeletal metastases (all P<0.05). Conclusions:The prognosis is extremely poor in HR-NB patients with multiple skeletal metastases at the initial diagnosis.The amplification of MYCN and the viscerocranium metastasis are the poor prognostic factors for HR-NB patients with skeletal metastases.

Astragalosides are the main active constituents of traditional Chinese medicine Huang-Qi, of which cycloastragenol-type glycosides are the most typical and major bioactive compounds. This kind of compounds exhibit various biological functions including cardiovascular protective, neuroprotective, etc. Owing to the limitations of natural sources and the difficulties encountered in chemical synthesis, re-engineering of biosynthetic machinery will offer an alternative and promising approach to producing astragalosides. However, the biosynthetic pathway for astragalosides remains elusive due to their complex structures and numerous reaction types and steps. Herein, guided by transcriptome and phylogenetic analyses, a cycloartenol synthase and four glycosyltransferases catalyzing the committed steps in the biosynthesis of such bioactive astragalosides were functionally characterized from Astragalus membranaceus. AmCAS1, the first reported cycloartenol synthase from Astragalus genus, is capable of catalyzing the formation of cycloartenol; AmUGT15, AmUGT14, AmUGT13, and AmUGT7 are four glycosyltransferases biochemically characterized to catalyze 3-O-xylosylation, 3-O-glucosylation, 25-O-glucosylation/O-xylosylation and 2'-O-glucosylation of cycloastragenol glycosides, respectively. These findings not only clarified the crucial enzymes for the biosynthesis and the molecular basis for the structural diversity of astragalosides in Astragalus plants, also paved the way for further completely deciphering the biosynthetic pathway and constructing an artificial pathway for their efficient production.

Docynia longiunguis is a plant uniquely present in China and is of high edible and medicinal value. The analysis of its chloroplast genome will help clarify the phylogenetic relationship among Docynia and facilitate the development and utilization of D. longiunguis resources. Based on the alignment of chloroplast genome sequences of related species, the phylogeny and codon preference were analyzed. The total length of D. longiunguis chloroplast genome sequence was 158 914 bp (GenBank accession number is MW367027), with an average GC content of 36.7%. The length of the large single-copy (LSC), the small single-copy (SSC), and inverted repeats (IRs) are 87 020 bp, 19 156 bp, and 26 369 bp, respectively. A total of 102 functional genes were annotated, including 72 protein-coding genes, 26 tRNA genes, and 4 rRNA genes. The best model for constructing phylogenetic tree was TVM+F+R2. D. longiunguis and Docynia indica were clustered into a single group, while Docynia and Malus were clustered into a single group. Comparison of the chloroplast genome sequences of D. longiunguis and its five related species revealed that trnY (GUA)-psbD, ndhC-trnV (UAC), accD-psaI, psbZ-trnfM (CAU), ndhF-trnL gene regions varied greatly. The nucleic acid diversity analysis showed that there were 11 high variation areas with nucleotide variability > 0.01, all were located in the LSC and SSC regions. Except for D. longiunguis, the trnH genes in other sequences were located at the IRs/LSC junction and did not cross the boundary. Codon preference analysis showed that D. longiunguis chloroplast genome has the largest number of isoleucine (Ile) codons, up to 1 205. D. longiunguis has the closest genetic relationship with Malus baccata, Malus sieboldii, Malus hupehensis and Chaenomeles sinensis. Its chloroplast genome codon prefers to end with A/T. The chloroplast genome of D. longiunguis and other Rosaceae chloroplast genomes showed great differences in gene distribution in four boundary regions, while relatively small differences from the chloroplast genomes of Docynia delavayi and D. indica of the same genus were observed. The genome annotation, phylogenetic analysis and sequence alignment of chloroplast genome of D. longiunguis may facilitate the identification, development and utilization of this species.
Codon Usage ; Genome, Chloroplast ; Genomics ; Phylogeny ; Rosaceae