Aim To investigate the impact of Cinobu-facini on the proliferation,invasion,and apoptosis of HepG2 cells and the underlying mechanism.Methods The proliferation of HepG2 cells was assessed using the CCK-8 method following treatment with Cinobufaci-ni.The invasion capability of HepG2 cells was evalua-ted through Transwell assay after exposure to Cinobufa-cini.The apoptosis rates of HepG2 cells post Cinobufa-cini intervention were measured using flow cytometry,and the expression levels of VEGF in the culture medi-um of HepG2 cells were determined using enzyme-linked immunoassay.Furthermore,qRT-PCR and Western blot analyses were conducted to assess the im-pact of Cinobufacini on mRNA and protein expression levels related to the CXCL5/FOXD1/VEGF pathway.The interaction between CXCL5 and FOXD1 was inves-tigated via co-immunoprecipitation.Results Cinobufa-cini treatment led to a gradual decrease in HepG2 cell viability in a dose-dependent manner compared to the control group(P＜0.05).Moreover,Cinobufacini sig-nificantly suppressed HepG2 cell invasion(P＜0.05)while enhancing cell apoptosis(P＜0.05).Notably,Cinobufacini exhibited inhibitory effects on the CX-CL5/FOXD1/VEGF pathway,as evidenced by re-duced expression of related mRNA and proteins(P＜0.05).FOXD1 was identified as the binding site of CXCL5.Overexpression of CXCL5 resulted in in-creased proliferation and VEGF secretion by HepG2 cells(P＜0.05),and increased expression of FOXD1 and VEGF(P＜0.05).However,Cinobufacini inter-vention effectively inhibited liver cancer cell prolifera-tion and invasion(P＜0.05),promoted apoptosis(P＜0.05),reduced VEGF secretion by HepG2 cells(P＜0.05),and downregulated the expression of CXCL5 and FOXD1 in HepG2 cells(P＜0.05);but com-pared with the unexpressed group of Cinobufacini,its ability to inhibit cell activity was weakened(P＜0.05),and its ability to inhibit the expression of CX-CL5,FOXD1,and VEGF was weakened(P＜0.05).Conclusion Cinobufacini may inhibit HepG2 cell pro-liferation and invasion and promote HepG2 cell apopto-sis by regulating the CXCL5/FOXD1/VEGF pathway.

Purpose::Ischemia and hypoxia are the main factors limiting limb replantation and transplantation. Static cold storage (SCS), a common preservation method for tissues and organs, can only prolong limb ischemia time to 4 - 6 h. The normothermic machine perfusion (NMP) is a promising method for the preservation of tissues and organs, which can extend the preservation time in vitro by providing continuous oxygen and nutrients. This study aimed to evaluate the difference in the efficacy of the 2 limb preservation methods. Methods::The 6 forelimbs from beagle dogs were divided into 2 groups. In the SCS group ( n = 3), the limbs were preserved in a sterile refrigerator at 4 °C for 24 h, and in the NMP group ( n = 3), the perfusate prepared with autologous blood was used for the oxygenated machine perfusion at physiological temperature for 24 h, and the solution was changed every 6 h. The effects of limb storage were evaluated by weight gain, perfusate biochemical analysis, enzyme-linked immunosorbent assay, and histological analysis. All statistical analyses and graphs were performed using GraphPad Prism 9.0 one-way or two-way analysis of variance. The p value of less than 0.05 was considered to indicate statistical significance. Results::In the NMP group, the weight gained percentage was 11.72% ± 4.06%; the hypoxia-inducible factor-1α contents showed no significant changes; the shape of muscle fibers was normal; the gap between muscle fibers slightly increased, showing the intercellular distance of (30.19 ± 2.83) μm; and the vascular α-smooth muscle actin (α-SMA) contents were lower than those in the normal blood vessels. The creatine kinase level in the perfusate of the NMP group increased from the beginning of perfusion, decreased after each perfusate change, and remained stable at the end of perfusion showing a peak level of 4097.6 U/L. The lactate dehydrogenase level of the NMP group increased near the end of perfusion and reached the peak level of 374.4 U/L. In the SCS group, the percentage of weight gain was 0.18% ± 0.10%, and the contents of hypoxia-inducible factor-1α increased gradually and reached the maximum level of (164.85 ± 20.75) pg/mL at the end of the experiment. The muscle fibers lost their normal shape and the gap between muscle fibers increased, showing an intercellular distance of (41.66 ± 5.38) μm. The contents of vascular α-SMA were much lower in the SCS group as compared to normal blood vessels.Conclusions::NMP caused lesser muscle damage and contained more vascular α-SMA as compared to SCS. This study demonstrated that NMP of the amputated limb with perfusate solution based on autologous blood could maintain the physiological activities of the limb for at least 24 h.

Objective: To evaluate the clinical characteristics and prognostic factors of patients with Philadelphia-negative myeloproliferative neoplasm-accelerated phase/blast phase (MPN-AP/BP) . Methods: A total of 67 patients with MPN-AP/BP were enrolled from February 2014 to December 2021 at the Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences. Their clinical features and prognostic factors were analyzed retrospectively. Results: ① Sixty-seven patients with MPN-AP/BP with a median age of 60 (range, 33-75) years, including 31 males (46.3% ) and 36 females (53.7% ) , were analyzed. Forty-eight patients progressed from primary myelofibrosis (PMF) , and 19 progressed from other myeloproliferative neoplasms (MPNs) , which included polycythemia vera, essential thrombocythemia, and MPN unclassifiable. Patients who progressed from PMF had higher lactate dehydrogenase (LDH) levels than those who progressed from other MPNs (925.95 vs. 576.2 U/L, P=0.011) , and there were higher proportions of patients who progressed from PMF with splenomegaly (81.4% vs. 57.9% , P=0.05) , a myelofibrosis grade of ≥2 (93.6% vs. 63.2% , P=0.004) , and a shorter duration from diagnosis to the transformation to AP/BP (28.7 vs. 81 months, P=0.001) . ② JAK2V617F, CALR, and MPLW515 were detected in 41 (61.2% ) , 13 (19.4% ) , and 3 (4.5% ) patients, respectively, whereas 10 (14.9% ) patients did not have any driver mutations (triple-negative) . Other than driver mutations, the most frequently mutated genes were ASXL1 (42.2% , n=27) , SRSF2 (25% , n=16) , SETBP1 (22.6% , n=15) , TET2 (20.3% , n=13) , RUNX1 (20.3% , n=13) , and TP53 (17.2% , n=11) . The ASXL1 mutation was more enriched (51.1% vs. 21.1% , P=0.03) , and the median variant allele fraction (VAF) of the SRSF2 mutation (median VAF, 48.8% vs. 39.6% ; P=0.008) was higher in patients who progressed from PMF than those who progressed from other MPNs. ③ In the multivariate analysis, the complex karyotype (hazard ratio, 2.53; 95% confidence interval, 1.06-6.05; P=0.036) was independently associated with worse overall survival (OS) . Patients who received allogeneic stem cell transplantation (allo-HSCT) (median OS, 21.3 vs. 3 months; P=0.05) or acute myeloid leukemia-like (AML-like) therapy (median OS, 13 vs. 3 months; P=0.011) had significantly better OS than those who received supportive therapy. Conclusion: The proportions of patients with PMF-AP/BP with splenomegaly, myelofibrosis grade ≥2, a higher LDH level, and a shorter duration from diagnosis to the transformation to AP/BP were higher than those of patients with other Philadelphia-negative MPN-AP/BP. The complex karyotype was an independent prognostic factor for OS. Compared with supportive therapy, AML-like therapy and allo-HSCT could prolong the OS of patients with MPN-AP/BP.
Male ; Female ; Humans ; Adult ; Middle Aged ; Aged ; Blast Crisis/drug therapy* ; Primary Myelofibrosis/genetics* ; Prognosis ; Splenomegaly ; Retrospective Studies ; Myeloproliferative Disorders/genetics* ; Mutation ; Leukemia, Myeloid, Acute ; Janus Kinase 2/genetics*

Objective: To compare clinical and laboratory features between JAK2 exon12 and JAK2 V617F mutated polycythemia vera (PV) . Method: We collected data from 570 consecutive newly-diagnosed subjects with PV and JAK2 mutation, and compared clinical and laboratory features between patients with JAK2 exon12 and JAK2 V617F mutation. Results: 543 (95.3%) subjects harboured JAK2 V617F mutation (JAK2 V617F cohort) , 24 (4.2%) harboured JAK2 exon12 mutations (JAK2 exon12 cohort) , and 3 (0.5%) harboured JAK2 exon12 and JAK2 V617F mutations. The mutations in JAK2 exon12 including deletion (n=10, 37.0%) , deletion accompanied insertion (n=10, 37.0%) , and missense mutations (n=7, 25.9%) . Comparing with JAK2 V617F cohort, subjects in JAK2 exon12 cohort were younger [median age 50 (20-73) years versus 59 (25-91) years, P=0.040], had higher RBC counts [8.19 (5.88-10.94) ×10(12)/L versus 7.14 (4.11-10.64) ×10(12)/L, P<0.001] and hematocrit [64.1% (53.7-79.0%) versus 59.6% (47.2%-77.1%) , P=0.001], but lower WBC counts [8.29 (3.2-18.99) ×10(9)/L versus 12.91 (3.24-38.3) ×10(9)/L, P<0.001], platelet counts [313 (83-1433) ×10(9)/L versus 470 (61-2169) ×10(9)/L, P<0.001] and epoetin [0.70 (0.06-3.27) versus 1.14 (0.01-10.16) IU/L, P=0.002] levels. We reviewed bone marrow histology at diagnosis in 20 subjects with each type of mutation matched for age and sex. Subjects with JAK2 exon12 mutations had fewer loose megakaryocyte cluster (40% versus 80%, P=0.022) compared with subjects with JAK2 V617F. The median follow-ups were 30 months (range 4-83) and 37 months (range 1-84) for cohorts with JAK2 V617F and JAK2 exon12, respectively. There was no difference in overall survival (P=0.422) and thrombosis-free survival (P=0.900) . Conclusions: Compared with patients with JAK2 V617F mutation, patients with JAK2 exon12 mutation were younger, and had more obvious erythrocytosis and less loose cluster of megakaryocytes.
Adult ; Aged ; Aged, 80 and over ; Bone Marrow/pathology* ; Exons ; Humans ; Janus Kinase 2/genetics* ; Middle Aged ; Mutation ; Mutation, Missense ; Polycythemia Vera/genetics* ; Young Adult

Objective: To analyze the course of disease and epidemiological parameters of COVID-19 and provide evidence for making prevention and control strategies. Methods: To display the distribution of course of disease of the infectors who had close contacts with COVID-19 cases from January 1 to March 15, 2020 in Guangdong Provincial, the models of Lognormal, Weibull and gamma distribution were applied. A descriptive analysis was conducted on the basic characteristics and epidemiological parameters of course of disease. Results: In total, 515 of 11 580 close contacts were infected, with an attack rate about 4.4%, including 449 confirmed cases and 66 asymptomatic cases. Lognormal distribution was fitting best for latent period, incubation period, pre-symptomatic infection period of confirmed cases and infection period of asymptomatic cases; Gamma distribution was fitting best for infectious period and clinical symptom period of confirmed cases; Weibull distribution was fitting best for latent period of asymptomatic cases. The latent period, incubation period, pre-symptomatic infection period, infectious period and clinical symptoms period of confirmed cases were 4.50 (95%CI:3.86-5.13) days, 5.12 (95%CI:4.63-5.62) days, 0.87 (95%CI:0.67-1.07) days, 11.89 (95%CI:9.81-13.98) days and 22.00 (95%CI:21.24-22.77) days, respectively. The latent period and infectious period of asymptomatic cases were 8.88 (95%CI:6.89-10.86) days and 6.18 (95%CI:1.89-10.47) days, respectively. Conclusion: The estimated course of COVID-19 and related epidemiological parameters are similar to the existing data.
COVID-19 ; Cohort Studies ; Contact Tracing ; Humans ; Incidence ; Prospective Studies

Gut microbiome sequencing studies have great potential to translate microbial analysis outcomes into human health research. Sequencing strategies of 16S amplicon and whole-metagenome shotgun (WMS) are two main methods in microbiome research with respective advantages. However, how sample heterogeneity, sequencers and library preparation protocols affect the sequencing reproducibility of gut microbiome needs further investigation. This study aims to provide a reference for the selection of sequencing technologies by comparing differences in microbial composition from different sampling sites. The results of three widely adopted sequencers showed that the technical repetition correlation (r= 0. 94) was high in WMS method, while the biological repetition correlation (r = 0. 69) was low. Bray-Curtis distance identified that dissimilarity from biological replicates was larger than that of technical replicates (P<0. 001). In addition, dissimilarity and specific taxonomic profiles were observed between 16S and WMS datasets. Our results imply that homogenization is a necessary step before sample DNA extraction. The sequencers contributed less to taxonomic variation than the library preparation protocols. We developed an empirical Bayes approach that " borrowed information" in calculations and analyzed batch effect parameters using standardized data and prior distributions of (non-) parameters, which may improve population comparability between 16S and WMS and provide a basis for further application to fusion analysis of published 16S and microbial datasets.

Objective:To explore the influence of intensive analgesia on the incidence of post-traumatic stress disorder (PTSD) in acute trauma patients, and to develop new ideas for the prevention and treatment of PTSD.Methods:From January 2018 to November 2019, a prospective study was conducted on trauma patients who visited the Emergency Center of Affiliated Hospital of Xuzhou Medical University and met the enrollment criteria. The patients were divided into the intensive analgesia group (< 4) and non-intensive analgesia group (≥ 4) according to the mean pain score in 30 days. The epidemiological data, trauma-related parameters, analgesic schemes, VAS score, PCL-5 score, HADS score and incidence of PTSD of enrolled patients were collected. Appropriate statistical methods were used to analyze differences among the indicators between the two groups.Results:Eighty-four acute trauma cases were included in the study, 39 cases in the intensive analgesia group and 45 in the non-intensive analgesia group. There was no significant difference in baseline data between the two groups (all P>0.05). The incidence rate of PTSD and PCL-5 score of patients in the intensive analgesia group were all significantly lower than those in the non-intensive analgesia group in 1 month after the trauma (all P< 0.05). The HADS anxiety and depression scores of patients in the intensive analgesic group were significantly lower than those in the non-intensive analgesic group (all P< 0.05). All the analgesics were converted into the dosage of dezocine for comparison. The total dosage of analgesics (dezocine) used in patients of the intensive analgesia group was significantly higher than that in the non-intensive analgesia group within 30 days after injury ( P< 0.05). Conclusion:In the acute trauma patients, intensive analgesia after trauma can significantly reduce the incidence of PTSD as well as improve anxiety and depression symptoms.

Objective:To investigate the relationship between the changes in inflammatory markers levels and the onset of post-traumatic stress disorder (PTSD) in the early stage of acute trauma..Methods:From January 2018 to June 2020, patients with acute trauma who were admitted to the Affiliated Hospital of Xuzhou Medical University were selected as subjects. Peripheral venous blood was collected on admission, on the 3rd and 7th day after trauma for routine blood test, C-reactive protein (CRP) and procalcitonin (PCT). The neutrophil to lymphocyte ratio (NLR) was calculated. The PCL-5 scale was used to evaluate PTSD symptoms one month later. The patients were divided into the PTSD group and non-PTSD group with the score of 38 as the boundary. The change rule of NLR in the PTSD group and the non-PTSD group were analyzed.Results:Ninety-one trauma patients were enrolled, including 23 patients in the PTSD group and 68 patients in the non-PTSD group. Compared with the healthy control group, the NLR of 91 trauma patients on admission, on the 3rd and 7th day were significantly higher (all P< 0.01). The NLR of the PTSD group was increased on the 7th day after trauma, which was significantly higher than that of the non-PTSD group ( P= 0.025). The non-PTSD group showed a decreasing trend, of which NLR on the 7th day was significantly lower than that on admission ( P= 0.001). In addition, high level of NLR on the 7th day after trauma (β= 0.206, P= 0.01) was a risk factor for PTSD onset. Conclusions:Dynamic monitoring of the changes in NLR after acute trauma would be of great clinical value to early warning of PTSD.

Objective:To investigate the current management of nosocomial infection at medical institutions of all levels in Changzhou, so as to provide basis for standardizing nosocomial infections control of hospitals within a medical alliance.Methods:An electronic questionnaire was customized for online survey of 91 hospitals affiliated to eight regional medical alliances in Changzhou city in March 2019. The survey covered such aspects as general conditions of the hospital, profile of nosocomial infection control administrators and other staffing, supervision of hospital nosocomial infection programs, and training needs, as well as outstanding problems and suggestions.Frequency number and percentage represent enumeration data, and χ2 test was used to analyze the in-group differences of medical institutions of three levels. Results:Tertiary public hospitals were superior to the secondary and primary hospitals in organizational structure, professional staffing and target monitoring, with the differences of statistical significance( P<0.05). The most urgent training needs of medical institutions at all levels were knowledge in determination and reporting of infectious diseases/nosocomial infection/infection outbreaks; top imperatives and recommendations were development of operation rules for primary medical institutions and standardization of workflows. Conclusions:Staff of primary medical institutions need capacity building in nosocomial infection control; primary hospitals are equipped with incomplete nosocomial infection control information platform; key departments in general lack homogenous management. Tertiary hospitals are encouraged to play leadership in medical alliances in achieving standardized, homogenous and informationized nosocomial infection control within the medical alliances.

Objective:To investigate the effect of post-traumatic pain on post-traumatic stress disorder (PTSD).Methods:A prospective trial was conducted with the recruitment of patients referred to the Emergency Center of Affiliated Hospital of Xuzhou Medical University for acute trauma from January 2018 to June 2019, with the exclusion criteria: age < 18 years, severe eye injury, severe craniocerebral injury and other critical conditions. The clinical data and written informed consent were collected at admission, the post-traumatic pain was assessed by VAS scores and the trauma severity was assessed by ISS score. APACHEⅡ score were evaluated within 24 h after admission. One month after trauma, patients with history of mental illness, or history of major psychogenic trauma within one year or drug addiction were further excluded. Accordingly, 64 eligible patients were evaluated by VAS, and the PCL-5 scale was used to evaluated their PTSD symptoms. The patients were divided into the PTSD group and non-PTSD group according to PCL-5 score≥ 38, and the difference between the two groups in post-traumatic VAS scores was compared. Logistic regression analysis was used to analyze the risk factors of PTSD. Spearman correlation analysis was adopted to establish the correlation between VAS score at admission and PCL-5 score at one month after trauma. The receiver operator characteristic (ROC) curve was used to analyze the predictive value of traumatic pain intensity for PTSD onset.Results:Sixty-four patients were selected and defined as the PTSD group ( n=19) and non-PTSD group ( n=45). The VAS score at admission was significantly higher in the PTSD group than that in the non-PTSD group ( P=0.006). There was no significant difference in VAS scores and VAS variations at 1 month, and in ISS scores at admission and APACHEⅡ scores within 24 h after admission between the two groups. Traumatic pain was an independent risk factor for PTSD ( P=0.043). VAS score at admission was positively correlated with PCL-5 score at 1 month post-traumatically ( r=0.355, P=0.004). In addition, ROC curve analysis showed that VAS score > 8 at admission had predictive value for PTSD (sensitivity=100%, specificity=33%, P=0.000 2). Conclusions:Post-traumatic severe pain is an independent risk factor for PTSD, which requires prompt medical intervention.