ObjectiveTo explore the changes in volatile components， total polysaccharides， enzyme activity， and chromaticity value of Myristicae Semen Koji（MSK） during the fermentation process， and conduct correlation analysis. MethodsBased on gas chromatography-mass spectrometry（GC-MS）， the changes of volatile components in MSK at different fermentation times were identified. The phenol sulfuric acid method， dinitrosalicylic acid method（DNS）， and carboxymethyl cellulose sodium salt method（CMC-Na） were used to investigate the total polysaccharide content， amylase activity， and cellulase activity during the fermentation process. Visual analysis technology was used to explore the changes in chromaticity values， revealing the fermentation process of MSK and the dynamic changes of various measurement indicators， partial least squares-discriminant analysis（PLS-DA） was used to explore the differential compounds of MSK at different fermentation degrees， and Pearson correlation analysis was used to explore the correlation between volatile components of MSK and total polysaccharides， enzyme activity， and chromaticity values. ResultsA total of 60 volatile compounds were identified from MSK， the relative contents of components such as （+）-α-pinene， β-phellandrene， β-pinene， （+）-limonene， and p-cymene obviously increased， while the relative contents of components such as safrole， methyl isoeugenol， methyleugenol， myristicin， and elemicin significantly decreased. During the fermentation process， the total polysaccharide content showed an upward trend， while the activities of amylase and cellulase showed an initial increase followed by a decrease， and reached their maximum value at 40 h. the overall brightness（L^*） and total color difference（ΔE^*） gradually increased， while the changes in red-green value（a^*） and yellow-blue value（b^*） were not obvious. PLS-DA results showed that MSK could be clearly distinguished at different fermentation times， and 13 differential biomarkers were screened out. Pearson correlation analysis results showed that the contents of α-terpinene， β-phellandrene， methyleugenol， β-cubebene and myristic acid had an obvious correlation with chromaticity values. ConclusionAfter fermentation， the volatile components， total polysaccharides， amylase activity， and cellulase activity of MSK undergo significant changes， and there is a clear correlation between them and chromaticity values， which reveals the dynamic changes in the fermentation process and related indicators of MSK， laying a foundation for the quality control.

ObjectiveTo evaluate the effect and safety of Jiuxin Pill （救心丸） on exercise tolerance and quality of life in patients with stable angina pectoris （SAP）. MethodsA randomised， double-blind， placebo-controlled， multicentre study design was used to enroll 170 patients of SAP from nine centres， which were divided into 85 patients each in the trial group and control group with 1∶1 ratio. Both groups maintained the original western medicine treatment plan， and added Jiuxin Pill or placebo respectively， 2 pills （0.05 g） each time twicely for 28 days. The main outcomes were total exercise time （TED） in the exercise treadmill test and Seattle Angina Questionnaire （SAQ） scores including physical limitation （PL）， angina stability （AS）， angina frequency （AF）， treatment satisfaction （TS）， and disease perception （DP）. The secondary outcomes were exercise treadmill test indicators including heart rate recovery in 1 min （HRR1）， metabolic equivalents （METs）， maximum magnitude of ST-segment depression， and the Borg rating of perceived exertion scale， the average number of angina attacks per week， withdrawal and reduction rate of nitroglycerin， traditional Chinese medicine syndrome scores， incidence of major adverse cardiovascular events. Safety indicators were evaluated and the occurrence of adverse events during the trial was recorded. Data was collected before treatment， day 28±2 in treatment period， and follow-up at day 56 which is 28±2 days after treatment period finished. ResultsEighty-four and eighty-five patients respectively from trial group and control group were included to the full analysis set （FAS） and safety analysis set （SS）. Compared with the group before treatment and with the control group after treatment， the trial group had higher TED， HRR1， and METs， and lower maximum magnitude of ST-segment depression and Borg rating of perceived exertion scores after treatment （P＜0.01）. Compared with the group before treatment and with the control group after treatment and at follow-up， the total SAQ score and scores of AS， AF， TS and DP of the trial group after treatment and at follow-up elevated， while the average number of angina attacks per week and traditional Chinese medicine syndrome scores reduced （P＜0.01）. There was no statistically significant difference in the withdrawal and reduction rate of nitroglycerin between groups （P＞0.05）. Major adverse cardiovascular events occurred in 1 case （1/84， 1.19%） in the trial group and 1 case （1/85， 1.18%） in the control group， and the difference between groups was not statistically significant （P＞0.05）. A total of 3 cases of adverse events occurred in the trial group （3/84， 3.57%）， and a total of 6 cases of adverse events occurred in the control group （6/85， 7.06%）， and there was no statistically significant difference in the incidence of adverse events between groups （P＞0.05）. ConclusionIn the treatment of SAP， Jiuxin Pill combined with conventional western medicine can further enhance exercise tolerance， improve quality of life， and demonstrate great safety.

Objective:To analyze the effects of SARS-CoV-2 infection and vaccination on virus mutation.Methods:The whole genome sequencing sequences of 2 659 local SARS-CoV-2 specimens from Jiangsu Province in 2023 were selected for analysis, and relevant information such as demographic and clinical characteristics were collected, and the effects of infection and vaccination on the genome-wide mutation rate and S gene′s selective pressure of the virus were analyzed by univariate and multivariate linear regression models.Results:The average age of these infected patients was 55.0 (31.0, 74.0) years, 1 150 cases (43.2%) in the age group of ≥60 years, 1 367 cases (51.4%) were males, 2 044 cases (76.9%) had a history of COVID-19 vaccination, and 1 629 cases (61.3%) had the first-time infection. The clinical symptoms of the infected patients were mainly mild, with a total of 2434 cases (91.5%), and 29 cases (1.1%) with severe symptoms or more. The average substitution rate of SARS-CoV-2 was 9.69 (9.38, 9.98)×10 -4 subs/site/year, and the dN/dS value of the S gene was 6.08 (5.56, 8.66), which was significantly greater than that of 1 ( P<0.001), indicating positive selection. The result of univariate and multivariate linear regression model analysis showed that the SARS-CoV-2 substitution rate was higher in those with vaccination history and reinfection, aged 20-30 years, ≥60 years, and the SARS-CoV-2 substitution rate was lower in males with moderate clinical symptoms and severe disease and above. Those with a history of vaccination and reinfection, aged 50-60 years old, ≥60 years old have smaller S gene dN/dS. Conclusions:Under the immune pressure exerted by vaccination and infection, the genome-wide mutation of SARS-COV-2 accelerated, but the non-synonymous mutation rate of the S gene decreased. The mechanism causing these phenomena needs further study.

Objective:To explore the correlation between the levels of serum lipoxin A4 (LXA4), cysteine-rich protein 61 (Cyr61) and disease severity in patients with knee osteoarthritis.Methods:A total of 106 patients with knee osteoarthritis treated in the Xianning Central Hospital (the First Affiliated Hospital of Hubei University of Science and Technology) from October 2017 to October 2019 were selected as the study subjects, and the patients were divided into mild to moderate group (63 cases) and severe group (43 cases) according to the Kellgren-Lawrence (K-L) grading criteria. Meanwhile 80 healthy subjects who underwent physical examination during the same period were selected as the control group. The levels of serum LXA4 and Cyr61 were detected by enzyme-linked immunosorbent assay. Spearman correlation was used to analyze the correlation between serum LXA4 and Cyr61 levels and the severity of knee osteoarthritis. Logistic regression was used to analyze the influencing factors of the severity of knee osteoarthritis. Receiver operative characteristic (ROC) curve was used to analyze the predictive efficacy of serum LXA4 and Cyr61 in patients with severe knee osteoarthritis.Results:The serum LXA4 level in the control group was (56.47 ± 9.62) μg/L , which was significantly higher than that in the mild to moderate group (46.16 ± 7.77) μg/L and the severe group (39.57 ± 6.20) μg/L, and the serum LXA4 level in the mild to moderate group was higher than that in the severe group, there were statistical differences ( P<0.05). The serum Cyr61 level in the control group was (25.07 ± 2.77) ng/L, which was significantly lower than that in the mild to moderate group (89.76 ± 10.47) ng/L and the severe group (96.88 ± 8.47) ng/L, and the serum LXA4 level in the mild to moderate group was lower than that in the severe group, there were statistical differences ( P<0.05). Spearman correlation analysis showed that the severity of disease in patients with knee osteoarthritis was negatively correlated with serum LXA4 level ( r = - 0.451, P<0.05) and positively correlated with serum Cyr61 level ( r = 0.358, P<0.05). The results of Logistic regression analysis showed that serum Cyr61 and LXA4 were predictors of the severity of knee osteoarthritis ( P<0.05). The area under the curve (AUC) of combination of serum LXA4 and Cyr61 in predicting severe knee osteoarthritis was 0.819, the sensitivity was 79.07%, and the specificity was 79.34%. Conclusions:The decrease of serum LXA4 level and the increase of Cyr61 level in patients with knee osteoarthritis are related to the severity of the disease. The combination of the two has the ability of clinical auxiliary diagnosis of the severity of osteoarthritis.

Objective:To investigate whether there are differences in disease outcomes among patients with Crohn′s disease (CD) L4 subtype.Methods:A total of 488 CD patients who were initially diagnosed at the Quzhou People′s Hospital and the Sir Run Run Shaw Hospital Affiliated to Zhejiang University School of Medicine from January 2017 to December 2019 were collected. They were divided into L4a, L4b, and non-L4 groups based on disease site classification, and the differences in intestinal complications and first abdominal surgery rates among the three groups were compared.Results:Among the 488 CD patients included in the study, 196(40.2%) were classified as L4 type, with L4b type being the predominant type (28.3%, 138/488). By confirming the time-dependent receiver operating characteristic (ROC) curve, the disease site can predict the surgical rate during follow-up of CD patients. Compared with non-L4 patients, L4b patients had a higher follow-up surgery rate [59.4%(76/128) vs 27.4%(80/292), P<0.001], while L4a patients had a lower rate [10.3%(6/58) vs 27.4%(80/292), P=0.009 6]. At the same time, the recurrence rate of intestinal obstruction in L4b patients during follow-up was significantly higher than that in L4a and non-L4 patients [46.9%(60/128) vs 6.9%(4/58), P<0.001; 46.9%(60/128) vs 11.6%(34/292), P=0.007]. The use of biologics can reduce the surgery rate in L4b patients ( HR: 1.93, 95% CI: 1.03-3.63, P=0.040 6). Conclusions:The incidence rate of L4b is high in newly diagnosed CD patients, and L4b is a high risk factor for poor prognosis of CD, so early use of biological agents is recommended. Accurate classification of disease sites can guide individualized clinical treatment.

Objective:To analyze the influence of serological indexes on the liver cirrhosis (LC) in patients with chronic hepatitis B, and to construct a nomogram model.Methods:The clinical data of 220 patients with chronic hepatitis B in Xianning Central Hospital from January 2019 to December 2021 were retrospectively analyzed. Among them, 42 patients developed LC (LC group), and 178 cases did not develop LC (non-LC group). The patient′s fasting peripheral venous blood was taken in the morning. The platelet, red blood cell, white blood cell, fasting blood glucose, alanine aminotransferase (ALT), aspartate aminotransferase (AST), triacylglycerol (TG), total cholesterol (TC), total bilirubin (TBiL), albumin, globulin, alkaline phosphatase (ALP), γ-glutamyltransferase (GGT), prothrombin time (PT), thrombin time (TT), D-dimer (D-D), alpha-fetoprotein (AFP) and C-reactive protein (CRP) were detected. Receiver operating characteristic (ROC) curve was used to analyze the efficacy of each index in predicting LC in patients with chronic hepatitis B. Multivariate Logistic regression was used to analyze the independent risk factors for LC in patients with chronic hepatitis B. The R software "rms" package was used to construct a nomogram model to predict the LC in patients with chronic hepatitis B, the correction curve was used to internally verify the prediction model, and the decision curve evaluated the efficacy of the prediction model.Results:The TBiL, ALP, GGT, PT, TT, D-D, AFP and CRP in LC group were significantly higher than those in non-LC group: (50.57 ± 5.61) μmol/L vs. (46.69 ± 3.92) μmol/L, (105.23 ± 30.60) U/L vs. (75.14 ± 26.45) U/L, (68.73 ± 19.47) U/L vs. (50.39 ± 14.21) U/L, (13.88 ± 1.98) s vs. (13.01 ± 2.10) s, (18.88 ± 2.56) s vs. (15.98 ± 2.43) s, (2.62 ± 1.04) mg/L vs. (1.34 ± 0.63) mg/L, (4.19 ± 1.95) μg/L vs. (2.66 ± 1.21) μg/L and (8.54 ± 1.22) mg/L vs. (7.47 ± 0.79) mg/L, the platelet, ALT, AST and albumin were significantly lower than those in the non-LC group: (129.63 ± 32.66) × 10 9/L vs. (183.53 ± 56.31) ×10 9/L, (131.27 ± 22.19) U/L vs. (157.57 ± 38.67) U/L, (112.76 ± 19.57) U/L vs. (125.16 ± 21.84) U/L and (29.79 ± 6.17) g/L vs. (33.52 ± 5.89) g/L, and there were statistical differences ( P<0.01 or <0.05); there were no statistical differences in red blood cell, white blood cell, fasting blood glucose, TG, TC and globulin between the two groups ( P>0.05). ROC curve analysis result showed that the area under the curve (AUC) of AFP, platelet, ALT, AST, ALP, GGT, TBiL, albumin, D-D, CRP, PT and TT for predicting LC in patients with chronic hepatitis B were 0.731, 0.798, 0.723, 0.676, 0.766, 0.762, 0.710, 0.673, 0.856, 0.759, 0.603 and 0.786, and the optimal cut-off values were 4.64 μg/L, 162.56 × 10 9/L, 155.67 U/L, 122.37 U/L, 95.17 U/L, 68.96 U/L, 49.95 μmol/L, 28.8 g/L, 1.64 mg/L, 8.55 mg/L, 12 s and 18 s. Multivariate Logistic regression analysis result showed that AFP (>4.64 μg/L), platelet (≤162.56 × 10 9/L), ALP (>95.17 U/L), GGT (>68.96 U/L), D-D (>1.64 mg/L) and TT (>18 s) were independent risk factors for LC in patients with chronic hepatitis B ( OR = 1.278, 1.428, 1.488, 1.356, 1.513 and 1.369; 95% CI 1.109 to 1.369, 1.269 to 1.623, 1.217 to 1.894, 1.127 to 1.669, 1.342 to 1.878 and 1.169 to 1.583; P<0.05 or <0.01). The AFP, platelet, ALP, GGT, D-D and TT were used as predictors to construct a nomogram model for predicting the LC in patients with chronic hepatitis B. The correction curve of the nomogram model to predict the LC in patients with chronic hepatitis B was close to the ideal curve (C-index was 0.739, 95% CI 0.615 to 0.876); the decision curve analysis result showed that the prediction model had higher clinical net benefit when the risk threshold > 0.26 than a single index, and that it had significantly additional clinical net benefit. Conclusions:The AFP, platelets, ALP, GGT, D-D and TT are independent risk factors for LC in patients with chronic hepatitis B, and the nomogram model constructed based on these factors could provide important guidance for the prevention and treatment of LC in patients with chronic hepatitis B.

Objective:To investigate the safety and efficacy of total pelvic exenteration (TPE) for treating late complications of radiation-induced pelvic injury.Methods:This was a descriptive case series study. The inclusion criteria were as follows: (1) confirmed radiation-induced pelvic injury after radiotherapy for pelvic malignancies; (2) late complications of radiation-induced pelvic injury, such as bleeding, perforation, fistula, and obstruction, involving multiple pelvic organs; (3) TPE recommended by a multidisciplinary team; (4) patient in good preoperative condition and considered fit enough to tolerate TPE; and (5) patient extremely willing to undergo the procedure and accept the associated risks. The exclusion criteria were as follows: (1) preoperative or intraoperative diagnosis of tumor recurrence or metastasis; (2) had only undergone diversion or bypass surgery after laparoscopic exploration; and (3) incomplete medical records. Clinical and follow-up data of patients who had undergone TPE for late complications of radiation-induced pelvic injury between March 2020 and September 2022 at the Sixth Affiliated Hospital of Sun Yat-sen University were analyzed. Perioperative recovery, postoperative complications, perioperative deaths, and quality of life 1 year postoperatively were recorded.Results:The study cohort comprised 14 women, nine of whom had recto-vagino-vesical fistulas, two vesicovaginal fistulas, one ileo-vesical fistula and rectal necrosis, one ileo-vesical and rectovaginal fistulas, and one rectal ulcer and bilateral ureteral stenosis. The mean duration of surgery was 592.1±167.6 minutes and the median blood loss 550 (100–6000) mL. Ten patients underwent intestinal reconstruction, and four the Hartmann procedure. Ten patients underwent urinary reconstruction using Bricker's procedure and 7 underwent pelvic floor reconstruction. The mean postoperative hospital stay was 23.6±14.9 days. Seven patients (7/14) had serious postoperative complications (Clavien-Dindo IIIa to IVb), including surgical site infections in eight, abdominopelvic abscesses in five, pulmonary infections in five, intestinal obstruction in four, and urinary leakage in two. Empty pelvis syndrome (EPS) was diagnosed in five patients, none of whom had undergone pelvic floor reconstruction. Five of the seven patients who had not undergone pelvic floor reconstruction developed EPS, compared with none of those who had undergone pelvic floor reconstruction. One patient with EPS underwent reoperation because of a pelvic abscess, pelvic hemorrhage, and intestinal obstruction. There were no perioperative deaths. During 18.9±10.1 months of follow-up, three patients died, two of renal failure, which was a preoperative comorbidity, and one of COVID-19. The remaining patients had gradual and significant relief of symptoms during follow-up. QLQ-C30 assessment of postoperative quality of life showed gradual improvement in all functional domains and general health at 1, 3, and 6 months postoperatively (all P<0.05). Conclusions:TPE is a feasible procedure for treating late complications of radiation-induced pelvic injury combined with complex pelvic fistulas. TPE is effective in alleviating symptoms and improving quality of life. However, the indications for this procedure should be strictly controlled and the surgery carried out only by experienced surgeons.

Objective:To investigate the safety and efficacy of total pelvic exenteration (TPE) for treating late complications of radiation-induced pelvic injury.Methods:This was a descriptive case series study. The inclusion criteria were as follows: (1) confirmed radiation-induced pelvic injury after radiotherapy for pelvic malignancies; (2) late complications of radiation-induced pelvic injury, such as bleeding, perforation, fistula, and obstruction, involving multiple pelvic organs; (3) TPE recommended by a multidisciplinary team; (4) patient in good preoperative condition and considered fit enough to tolerate TPE; and (5) patient extremely willing to undergo the procedure and accept the associated risks. The exclusion criteria were as follows: (1) preoperative or intraoperative diagnosis of tumor recurrence or metastasis; (2) had only undergone diversion or bypass surgery after laparoscopic exploration; and (3) incomplete medical records. Clinical and follow-up data of patients who had undergone TPE for late complications of radiation-induced pelvic injury between March 2020 and September 2022 at the Sixth Affiliated Hospital of Sun Yat-sen University were analyzed. Perioperative recovery, postoperative complications, perioperative deaths, and quality of life 1 year postoperatively were recorded.Results:The study cohort comprised 14 women, nine of whom had recto-vagino-vesical fistulas, two vesicovaginal fistulas, one ileo-vesical fistula and rectal necrosis, one ileo-vesical and rectovaginal fistulas, and one rectal ulcer and bilateral ureteral stenosis. The mean duration of surgery was 592.1±167.6 minutes and the median blood loss 550 (100–6000) mL. Ten patients underwent intestinal reconstruction, and four the Hartmann procedure. Ten patients underwent urinary reconstruction using Bricker's procedure and 7 underwent pelvic floor reconstruction. The mean postoperative hospital stay was 23.6±14.9 days. Seven patients (7/14) had serious postoperative complications (Clavien-Dindo IIIa to IVb), including surgical site infections in eight, abdominopelvic abscesses in five, pulmonary infections in five, intestinal obstruction in four, and urinary leakage in two. Empty pelvis syndrome (EPS) was diagnosed in five patients, none of whom had undergone pelvic floor reconstruction. Five of the seven patients who had not undergone pelvic floor reconstruction developed EPS, compared with none of those who had undergone pelvic floor reconstruction. One patient with EPS underwent reoperation because of a pelvic abscess, pelvic hemorrhage, and intestinal obstruction. There were no perioperative deaths. During 18.9±10.1 months of follow-up, three patients died, two of renal failure, which was a preoperative comorbidity, and one of COVID-19. The remaining patients had gradual and significant relief of symptoms during follow-up. QLQ-C30 assessment of postoperative quality of life showed gradual improvement in all functional domains and general health at 1, 3, and 6 months postoperatively (all P<0.05). Conclusions:TPE is a feasible procedure for treating late complications of radiation-induced pelvic injury combined with complex pelvic fistulas. TPE is effective in alleviating symptoms and improving quality of life. However, the indications for this procedure should be strictly controlled and the surgery carried out only by experienced surgeons.

The first rate-limiting enzyme of the serine synthesis pathway (SSP), phosphoglycerate dehydrogenase (PHGDH), is hyperactive in multiple tumors, which leads to the activation of SSP and promotes tumorigenesis. However, only a few inhibitors of PHGDH have been discovered to date, especially the covalent inhibitors of PHGDH. Here, we identified withangulatin A (WA), a natural small molecule, as a novel covalent inhibitor of PHGDH. Affinity-based protein profiling identified that WA could directly bind to PHGDH and inactivate the enzyme activity of PHGDH. Biolayer interferometry and LC-MS/MS analysis further demonstrated the selective covalent binding of WA to the cysteine 295 residue (Cys295) of PHGDH. With the covalent modification of Cys295, WA blocked the substrate-binding domain (SBD) of PHGDH and exerted an allosteric effect to induce PHGDH inactivation. Further studies revealed that with the inhibition of PHGDH mediated by WA, the glutathione synthesis was decreased and intracellular levels of reactive oxygen species (ROS) were elevated, leading to the inhibition of tumor proliferation. This study indicates WA as a novel PHGDH covalent inhibitor, which identifies Cys295 as a novel allosteric regulatory site of PHGDH and holds great potential in developing anti-tumor agents for targeting PHGDH.

Objective: : EID3 (EP300-interacting inhibitor of differentiation) was identified as a novel member of EID family and plays a pivotal role in colorectal cancer development. However, its role in glioma remained elusive. In current study, we identified EID3 as a novel oncogenic molecule in human glioma and is critical for glioma cell survival, proliferation and invasion. Methods: : A total of five patients with glioma were recruited in present study and fresh glioma samples were removed from patients. Four weeks old male non-obese diabetic severe combined immune deficiency (NOD/SCID) mice were used as transplant recipient models. The subcutaneous tumor size was calculated and recorded every week with vernier caliper. EID3 and AMP-activated protein kinase α1 (AMPKα1) expression levels were confirmed by real-time polymerase chain reaction and Western blot assays. Colony formation assays were performed to evaluate cell proliferation. Methyl thiazolyl tetrazolium (MTT) assays were performed for cell viability assessment. Trypan blue staining approach was applied for cell death assessment. Cell Apoptosis DNA ELISA Detection Kit was used for apoptosis assessment. Results: : EID3 was preferentially expressed in glioma tissues/cells, while undetectable in astrocytes, neuronal cells, or normal brain tissues. EID3 knocking down significantly hindered glioma cell proliferation and invasion, as well as induced reduction of cell viability, apoptosis and cell death. EID3 knocking down also greatly inhibited tumor growth in SCID mice. Knocking down of AMPKα1 could effectively rescue glioma cells from apoptosis and cell death caused by EID3 absence, indicating that AMPKα1 acted as a key downstream regulator of EID3 and mediated suppression effects caused by EID3 knocking down inhibition. These findings were confirmed in glioma cells generated patient-derived xenograft models. AMPKα1 protein levels were affected by MG132 treatment in glioma, which suggested EID3 might down regulate AMPKα1 through protein degradation. Conclusion : Collectively, our study demonstrated that EID3 promoted glioma cell proliferation and survival by inhibiting AMPKα1 expression. Targeting EID3 might represent a promising strategy for treating glioma.