ObjectiveTo establish a method for the identification of Haliotidis Concha and its counterfeits， and to improve its quality evaluation method. MethodsA total of 17 batches of Haliotis discus hannai， 4 batches of H. ruber， 3 batches of H. laevigata， 3 batches of H. ovina， 3 batches of H. diversicolor， 3 batches of H. asinina， 3 batches of H. iris were collected. Ultra-high performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry（UPLC-Q-Exactive-Orbitrap-MS/MS） was used to analyze the hydrolysates of different original Haliotidis Concha and its counterfeits， and the potential characteristic ions of each species were screened by Venn diagram. UPLC-triple quadrupole tandem mass spectrometry（UPLC-QqQ-MS/MS） was used to validate the characteristic ions， and the specific detection method of the characteristic ions was established. ResultsA total of 1 182， 167， 47， 89， 104， 203， 424 potential characteristic ions were screened from H. discus hannai， H. ruber， H. laevigata， H. ovina， H. diversicolor， H. asinina and H. iris， respectively. And 9 characteristic ions were selected. The precision， stability and repeatability of the 9 characteristic ions in the established identification method met the requirements. Different original Haliotidis Concha and its counterfeits could detect their own characteristic ions， including m/z 631.83-886.48（double charge） and m/z 631.83-443.74（double charge） of H. discus hannai， m/z 699.28-232.11（double charge） and m/z 699.28-544.27（double charge） of H. ruber， m/z 535.76-752.37（double charge） and m/z 535.76-548.28（double charge） of H. laevigata， m/z 708.35-442.28（double charge） and m/z 708.35-215.14（double charge） of H. ovina， m/z 561.33-614.86（triple charge）， m/z 561.33-468.28（triple charge）， m/z 608.29-618.32（double charge） and m/z 608.29-390.21（double charge） of H. diversicolor， m/z 769.85-274.10（double charge）， m/z 769.85-532.75（double charge）， m/z 827.43-646.36（single charge）， m/z 827.43-257.12（single charge） of H. asinina， and m/z 468.24-576.29（double charge） and m/z 468.24-505.26（double charge） of H. iris. ConclusionIn this study， a total of 9 characteristic ions are screened from 6 kinds of original Haliotidis Concha and its counterfeits， and a specific identification method is established， which is helpful to solve the limitations of the existing quality evaluation methods of Haliotidis Concha， and provide a basis for the production， circulation and medication quality.

ObjectiveTo establish a method for the identification of Haliotidis Concha and its counterfeits， and to improve its quality evaluation method. MethodsA total of 17 batches of Haliotis discus hannai， 4 batches of H. ruber， 3 batches of H. laevigata， 3 batches of H. ovina， 3 batches of H. diversicolor， 3 batches of H. asinina， 3 batches of H. iris were collected. Ultra-high performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry（UPLC-Q-Exactive-Orbitrap-MS/MS） was used to analyze the hydrolysates of different original Haliotidis Concha and its counterfeits， and the potential characteristic ions of each species were screened by Venn diagram. UPLC-triple quadrupole tandem mass spectrometry（UPLC-QqQ-MS/MS） was used to validate the characteristic ions， and the specific detection method of the characteristic ions was established. ResultsA total of 1 182， 167， 47， 89， 104， 203， 424 potential characteristic ions were screened from H. discus hannai， H. ruber， H. laevigata， H. ovina， H. diversicolor， H. asinina and H. iris， respectively. And 9 characteristic ions were selected. The precision， stability and repeatability of the 9 characteristic ions in the established identification method met the requirements. Different original Haliotidis Concha and its counterfeits could detect their own characteristic ions， including m/z 631.83-886.48（double charge） and m/z 631.83-443.74（double charge） of H. discus hannai， m/z 699.28-232.11（double charge） and m/z 699.28-544.27（double charge） of H. ruber， m/z 535.76-752.37（double charge） and m/z 535.76-548.28（double charge） of H. laevigata， m/z 708.35-442.28（double charge） and m/z 708.35-215.14（double charge） of H. ovina， m/z 561.33-614.86（triple charge）， m/z 561.33-468.28（triple charge）， m/z 608.29-618.32（double charge） and m/z 608.29-390.21（double charge） of H. diversicolor， m/z 769.85-274.10（double charge）， m/z 769.85-532.75（double charge）， m/z 827.43-646.36（single charge）， m/z 827.43-257.12（single charge） of H. asinina， and m/z 468.24-576.29（double charge） and m/z 468.24-505.26（double charge） of H. iris. ConclusionIn this study， a total of 9 characteristic ions are screened from 6 kinds of original Haliotidis Concha and its counterfeits， and a specific identification method is established， which is helpful to solve the limitations of the existing quality evaluation methods of Haliotidis Concha， and provide a basis for the production， circulation and medication quality.

Patients with overlap syndrome(OS)of autoimmune liver disease may present with more than one biochemical,immunological,histological or cholangiography features of autoimmune liver disease(AILD)and often require a combination of immunosuppressants for treatment.Pulmonary cryptococcosis is a type of invasive pneumomycosis caused by Cryptococcus neoformans or Cryptococcus gattii and has a relatively high incidence rate in immunocompromised patients.This case report presents a patient with OS who was found to have pulmonary cryptococcosis during immunosuppressive therapy and developed abnormal liver function during antifungal treatment.Based on the liver function of the patient,the feasibility of adjusting antifungal agents was assessed,and active treatment strategies for novel cryptococcal infection were developed under the close monitoring of liver function,which helped to avoid the progression of infection.It is suggested that before the initiation of immunosuppressive therapy,systemic foci of infection should be comprehensively evaluated,and suspicious foci of infection should be monitored continuously.

Objective:To investigate the expression of signal recognition particle 14(SRP14)in hepatocellular carcinoma(HCC)and its clinical significance.Methods:The data of SRP14 expression in HCC were obtained from bioinformatics study,and from investigation with quantitative reverse transcription polymerase chain reaction(qRT-PCR),immunohistochemical staining and Western blotting in clinical samples.The Kaplan-Meier analysis was used to determine the associations between SRP14 mRNA expression and the overall survival,progression-free survival,and disease-specific survival of HCC patients.The effect of SRP14 on the proliferation and migration of HCC cells were determined by EdU staining,MTS,Transwell and wound-healing assays.The potential mechanism for SRP14 regulating HCC was explored through Kyoto Encyclopedia of Genes and Genomes(KEGG)and Gene Ontology(GO)enrichment analysis as well as qRT-PCR.Results:According to the data from GSE14520,TNMplot database and clinical samples,compared with paired tumor-adjacent tissues,non-paired tumor-adjacent tissues and normal tissues,the mRNA expression of SPR14 in HCC tissues was upregulated(all P<0.05).In clinical samples,compared with paired tumor-adjacent tissues,the protein expression of SPR14 in HCC tissues was increased(P<0.05).The increased mRNA expression of SRP14 was associated with good overall survival,progression-free survival,and disease-specific survival in HCC patients.SRP14 inhibited the proliferation and migration of HCC cells in vitro.According to the KEGG and GO enrichment analysis,in non-specific HCC,the genes co-expressed with SRP14 may predominantly regulate protein synthesis,processing,and transport,while in nonalcoholic fatty liver disease related HCC,the genes co-expressed with SRP14 could control multiple signaling pathways such as MAPK,cAMP,PI3K-Akt,and Wnt.Mechanistically,SRP14 up-regulated the mRNA expression of tumor suppressor gene GPRC5A in HCC cells(P<0.05).Conclusion:SRP14 may regulate HCC progression and influence patient prognosis.

Objective:To compare the characteristics of background mucosa, lesion features, and the efficiency of endoscopic submucosal dissection(ESD)between elderly and non-elderly patients with early gastric cancer(EGC).Methods:This study retrospectively collected data on patients with EGC who underwent ESD treatment at Beijing Hospital from April 2020 to December 2022.The clinical characteristics, background mucosa, lesion features, ESD outcomes, and pathological results of the patients were analyzed to compare the differences between elderly and non-elderly patients.Results:A total of 100 patients with EGC were selected, comprising 57 patients in the elderly group and 43 patients in the non-elderly group, with a total of 111 lesions identified(64 lesions in the elderly group and 47 lesions in the non-elderly group).The proportion of patients with a history of chronic atrophic gastritis was significantly higher in the elderly group(89.5%、51/57)compared to the non-elderly group(74.4%、32/43), with a statistically significant difference( P=0.047).Additionally, the difference in the extent of atrophy between elderly patients with EGC and their non-elderly counterparts was statistically significant( P=0.022).Among these patients, the proportion of those classified as Kimura-Takemoto C0 to C1 in the elderly group(15.6%、10/64)was lower than that in the non-elderly group(40.4%、19/47).In contrast, the proportion of patients classified as C2 to C3 in the elderly group(65.6%、42/64)was higher than that in the non-elderly group(51.1%、24/47), and the proportion of those classified as O1 to O3 in elderly patients(12.5%、8/64)was also higher than in the non-elderly group(4.3%、2/47).Furthermore, the difference in the extent of intestinal metaplasia between elderly and non-elderly patients with early gastric cancer was statistically significant( P=0.007).The overall proportion of total intestinal metaplasia in elderly patients(85.9%、55/64)was significantly higher than that in non-elderly patients(61.7%、29/47).Notably, the proportion of patients exhibiting extensive intestinal metaplasia(intestinal metaplasia present in both the gastric antrum and gastric body)was greater in the elderly group(43.8%、28/64)compared to the non-elderly group(23.4%、11/47).The Kyoto gastric cancer risk endoscopic score for elderly patients with EGC was(2.43±1.28)points, significantly higher than that of the non-elderly group(1.72±1.41)points, with a statistically significant difference observed( t=2.778, P=0.006).No statistically significant differences were observed in the proportions of total resection rates, R0 resections, curative resections, or postoperative complications following ESD when comparing elderly patients with EGC to their non-elderly counterparts. Conclusions:The proportion of extensive atrophy and intestinal metaplasia was higher in the background mucosa of elderly patients with EGC, and correspondingly, the Kyoto endoscopic gastric cancer risk score was elevated.Therefore, endoscopic examinations for elderly patients with chronic atrophic gastritis should be conducted with greater care and comprehensiveness.

Objective:To investigate the risk factors for high-output stoma (HOS) in patients undergoing rectal cancer radical resection combined with terminal ileostomy.Methods:A retrospective analysis was conducted on the medical records of 104 patients who underwent radical resection for rectal cancer combined with terminal ileostomy at Northern Jiangsu People′s Hospital from January 2020 to December 2022. Among them, 77 were male and 27 were female, with ages ranging from 31 to 84 years and an average age of 62.60 years. HOS was defined as an average stoma output ≥1500 mL/24 h within 3 days after resuming oral intake. Based on the presence of HOS post-surgery, patients were divided into the HOS group ( n=29) and the non-HOS group ( n=77). The two groups were compared based on gender, age, body mass index (BMI), preoperative albumin, hemoglobin, fasting blood glucose, surgery duration, surgical approach, intraoperative blood loss, postoperative albumin, hemoglobin, white blood cell count, and C-reactive protein (CRP). Data were analyzed using SPSS27.0. Continuous data were expressed as mean±standard deviation( ± s), and independent t-tests or non-parametric tests were used for group comparisons. Chi-square tests were used for categorical data, and logistic regression was employed to evaluate the influence of various factors on the occurrence of HOS. Results:There were no statistically significant differences between the two groups in terms of gender, preoperative albumin, hemoglobin, fasting blood glucose, surgery duration, surgical approach, intraoperative blood loss, postoperative albumin, hemoglobin, or white blood cell count ( P>0.05). Univariate logistic regression analysis indicated that older patients were more likely to develop HOS ( P=0.037, OR=1.047), those with lower BMI were at higher risk of HOS ( P<0.001, OR=0.448), and elevated postoperative CRP was associated with an increased likelihood of HOS ( P<0.001, OR=1.027). Multivariate logistic regression analysis showed that BMI ( OR=0.302, 95% CI: 0.164-0.555, P<0.001) and postoperative CRP ( OR=1.045, 95% CI: 1.023-1.068, P<0.001) were independent risk factors for the occurrence of HOS. Conclusion:Lower BMI and elevated postoperative CRP are significant independent risk factors for the development of HOS in rectal cancer patients.

Pemphigus is a group of life-threatening chronic autoimmune bullous diseases. It is currently believed that the humoral immunity mediated by anti-desmoglein IgG antibodies is the primary mechanism underlying the occurrence of pemphigus. However, recent studies have indicated a critical role of various T cell subsets in the occurrence of pemphigus.

Objective:To isolate and identify SARS-CoV-2 epidemic strains and analyze the sequence characteristics of the virus strains following serial passages.Methods:Eleven nasopharyngeal swabs positive for SARS-CoV-2 antigen were collected from December 2022. Quantitative real-time PCR was used to detect SARS-CoV-2 nucleic acid, and positive specimens were inoculated onto Vero cells for virus isolation. The isolated strains were identified by Western blot and indirect immunofluorescence assay. The morphology of the isolated strains was observed using transmission electron microscopy. Nucleic acid was extracted from the isolates and passaged viruses for further sequencing and analysis.Results:All 11 specimens tested positive for SARS-CoV-2 using quantitative real-time PCR. SARS-CoV-2 strains were successfully isolated from seven specimens, and could be adaptively cultured, passaged, and expanded on Vero cells, achieving a peak titer exceeding 10 6.25 50% cell culture infectious dose (CCID 50)/ml. Western blot and indirect immunofluorescence results showed that the isolates could be specifically recognized by monoclonal antibodies and convalescent serum against SARS-CoV-2. Transmission electron microscopy revealed oval-shaped viral particles with diameters of approximately 100 nm. Next-generation sequencing of the viral isolates demonstrated a sequence homology greater than 99.50% with the Wuhan-Hu-1 reference strain (NC_045512) and 99.98% among the seven isolated strains, and all of the isolates belonged to the Omicron BF.7 variant. Sequence analysis after continuous passage and plaque purification of the BJ-NVSI-20230005 isolate showed that compared with passages 1-3, passages 4-6 had one nucleotide site mutation (C→T) in the ORF1ab gene and a deletion of 3 bp in the E gene, which resulted in a change from leucine to phenylalanine and the deletion of valine, respectively. Polymorphisms were observed in the sequences of plaque-purified clones. Conclusions:The seven successfully isolated SARS-CoV-2 strains all belong to the SARS-CoV-2 BF.7 variant, which is consistent with the prevalence trend in mainland China in December 2022.

Background: Long coronavirus disease 2019 (COVID-19) in recovered patients (RPs) is gradually recognized by more people. However, how long it will last and the underlining mechanism remains unclear. Methods: We conducted a prospective follow-up study to evaluate the long-term symptoms and clinical indices of RPs at one-year after discharge from Union Hospital, Wuhan, China between December 2020 to May 2021. We also performed the 16S rRNA sequencing of stool samples from RPs and healthy controls (HCs) and analyzed the correlation between the gut microbiota and long COVID-19. Results: In total, 187 RPs were enrolled, among them, 84 (44.9%) RPs reported long COVID-19 symptoms at one-year after discharge. The most common long-term symptoms were cardiopulmonary symptoms, including chest tightness after activity (39/187, 20.9%), palpitations on exercise (27/187, 14.4%), sputum (21/187, 11.2%), cough (15/187, 8.0%) and chest pain (13/187, 7.0%), followed by systemic symptoms including fatigue (34/187, 18.2%) and myalgia (20/187, 10.7%), and digestive symptoms including constipation (14/187, 7.5%), anorexia (13/187, 7.0%), and diarrhea (8/187, 4.3%). Sixty-six (35.9%) RPs presented either anxiety or depression (42/187 [22.8%] and 53/187 [28.8%] respectively), and the proportion of anxiety or depression in the long symptomatic group was significantly higher than that in the asymptomatic group (41/187 [50.6%] vs. 25/187 [24.3%]). Compared with the asymptomatic group, scores of all nine 36-Item Short Form General Health Survey domains were lower in the symptomatic group (all P < 0.05). One hundred thirty RPs and 32 HCs (non-severe acute respiratory syndrome coronavirus 2 infected subjects) performed fecal sample sequencing.Compared with HCs, symptomatic RPs had obvious gut microbiota dysbiosis including significantly reduced bacterial diversities and lower relative abundance of short-chain fatty acids (SCFAs)-producing salutary symbionts such as Eubacterium_hallii_group, Subdoligranulum, Ruminococcus, Dorea, Coprococcus, and Eubacterium_ventriosum_group. Meanwhile, the relative abundance of Eubacterium_hallii_group, Subdoligranulum, and Ruminococcus showed decreasing tendencies between HCs, the asymptomatic group, and the symptomatic group. Conclusion This study demonstrated the presence of long COVID-19 which correlates with gut microbiota dysbiosis in RPs at one-year after discharge, indicating gut microbiota may play an important role in long COVID-19.

The induction of hepatocyte-like cells (HLCs) in vitro is one of the effective ways to obtain a large number of useful hepatocyte, and these HLCs can be used in disease modeling, drug design, and toxicological evaluation. At present, the induction of HLCs in vitro is mainly achieved by introducing exogenous transcription factors, cytokines or small-molecule compounds. Since small-molecule compounds have the advantages of structural diversity, controllable time and dose, and convenient and safe operation, scientists are devoted to screening out the small-molecule compounds to replace exogenous transcription factors and cytokines, and such compounds have a promising application prospect in the field of regenerative medicine. This article reviews the studies on the in vitro induction of HLCs from pluripotent stem cells and other adult stem cells and summarizes the application of small-molecule compounds in the in vitro induction of HLCs, in order to provide ideas and references for the in vitro induction of HLCs.