Objective:To explore the predictive value of peripheral blood neutrophil to lymphocyte ratio (NLR) and serum trimethylamine oxide (TMAO) on in-hospital mortality events in patients with acute myocardial infarction complicated by cardiogenic shock (AMICS) upon admission.Methods:A retrospective collection of medical records of 103 AMICS patients admitted to the Shanxi Yingkang Life General Hospital from January 2018 to June 2023 was conducted. The patients were divided into a survival group ( n=78) and a death group ( n=25) based on whether they experienced in-hospital mortality events. Two groups of peripheral blood NLR, serum TMAO, baseline data, and other laboratory indicators were compared. A logistic regression model was used to analyze the risk factors for in-hospital mortality in AMICS patients and a prediction model was constructed. We established receiver operating characteristic (ROC) curves to evaluate the predictive efficacy of peripheral blood NLR, serum TMAO, peripheral blood NLR+ serum TMAO, and predictive model indicators for in-hospital mortality events in AMICS patients. Results:The peripheral blood NLR and serum TMAO levels in the death group were significantly higher than those in the survival group (all P<0.05). The age of the death group was higher than that of the survival group, and the proportion of hypertension, alanine aminotransferase, creatinine, random blood glucose, proportion of patients who did not receive emergency percutaneous coronary intervention (PCI) treatment, peak cardiac troponin I, B-type natriuretic peptide, Gensini score of coronary artery disease, and C-reactive protein were all higher than those of the survival group. Systolic blood pressure and platelet count were lower than those of the survival group, heart rate and erythrocyte sedimentation rate were faster than those of the survival group, and the pre hospital time was longer than that of the survival group. The differences were statistically significant (all P<0.05). The results of binary logistic regression analysis showed that after adjusting for confounding factors such as age, male proportion, body mass index, proportion of old myocardial infarction, proportion of hypertension, and proportion of PCI history, advanced age, long pre hospital time, failure to receive emergency PCI, elevated peripheral blood NLR, and elevated serum TMAO were independent risk factors for in-hospital mortality in AMICS patients ( P<0.05). The predictive model was obtained as 0.734×age+ 0.277×pre hospital time+ 2.263×failure to receive emergency PCI+ 0.549×peripheral blood NLR+ 0.608×serum TMAO-26.923. The ROC curve results showed that the area under the curve (AUC) values of peripheral blood NLR, serum TMAO, peripheral blood NLR+ serum TMAO, and predictive model indicators for predicting in-hospital mortality events in AMICS patients were 0.744, 0.781, 0.825, and 0.921, respectively. When the optimal cutoff value was taken, the sensitivities were 0.880, 0.520, 0.680, and 0.880, and the specificities were 0.526, 0.923, 0.872, and 0.821, respectively. Conclusions:Advanced age, long pre hospital duration, failure to undergo emergency PCI, elevated peripheral blood NLR, and elevated serum TMAO are independent risk factors for in-hospital mortality in AMICS patients. Peripheral blood NLR, serum TMAO, peripheral blood NLR+ serum TMAO, and prediction models all have certain predictive value for in-hospital mortality events in AMICS patients. Among them, the sensitivity and specificity of the prediction models are high, and the efficacy is good.

ObjectiveTo determine the optimal dose of Dachengqi Decoction （大承气汤， DCQD） for the treatment of acute intestinal obstruction （AIO） through a randomized， double-blind， dosage parallel controlled， multi-center clinical trial， and to providee evidence support for the reasonable dosage of DCQD in clinical practice. MethodsBased on the commonly used clinical dose of DCQD， three different groups were set up， including low-dose group which used Dahuang （Radix et Rhizoma Rhei） 12 g， Houpo （Cortex Magnoliae Officinalis） 9 g， Zhishi （Fructus Aurantii Immaturus） 9 g， and Mangxiao （Natrii Sulfas） 4.5 g， medium-dose group using Dahuang 36 g， Houpo 27 g， Zhishi 27 g， Mangxiao 13.5 g， and high-dose group using Dahuang 60 g， Houp0 45 g， Zhishi 45 g and Mangxiao 22.5 g. Initially， 149 AIO patients with Yangming （阳明） bowel excess syndrome were randomly assigned to three groups using a stratified randomization method， and both the patients and the doctors were blinded. In addition to conventional western medicine treatment， each group was given 12 bags of granules made from the raw herbs of DCQD at different doses， taken orally or injected through a gastric catheter once every 6 hours， 3 bags each time， for 3 consecutive days. After treatment， the indicators of the three groups of patients with primary AIO and non-primary AIO were evaluated respectively， and the full analysis set （FAS） and per-protocol set （PPS） were used for analysis. The primary outcomes were the time to recover voluntary bowel movements and voluntary flatulence. The secondary outcomes were the ideal rate of spontaneous defecation and the ideal rate of spontaneous flatus. The occurrence of adverse events during the study was recorded and analyzed using the safety analysis set （SS）. ResultsA total of 91 patients with primary AIO and 58 patients with non-primary AIO were included in the FAS and SS analysis， while 80 primary AIO patients and 56 non-primary AIO patients were included in the PPS analysis. Both FAS and PPS analysis showed significant differences in the time to recover voluntary bowel movements and voluntary flatulence among primary AIO patients in different dose groups of DCQD （P＜0.01）， and the high- and medium-dose groups assumed less time than the low-dose group （P＜0.05）. There was no statistically significant difference in the ideal rate of spontaneous defecation and spontaneous flatus among the three groups （P＞0.05）. And consistent results were seen in the non-primary AIO patients among the three groups. Five adverse events occurred in primary AIO patients （3 in the low-dose group， 1 in the medium-dose group， and 1 in the high-dose group）， mainly manifested as abdominal distension and abdominal pain， and there was no statistically significant difference in the incidence of adverse events （P＞0.05）. No adverse events occurred in patients with non-primary AIO. ConclusionDCQD， as an effective treatment for patients with AIO， is commonly used at a medium dose for patients with primary AIO and at a high dose for patients with non-primary AIO. The therapeutic advantage is mainly reflected in the shorter time to recover spontaneous defecation and spontaneous flatulence and the improvement of intestinal function.

The aim of this study was to produce recombinant porcine interferon gamma (rPoIFN-γ) by Chinese hamster ovarian (CHO) cells expression system and to analyze its antiviral activity. Firstly, we constructed the recombinant eukaryotic expression plasmid pcDNA3.1-PoIFN-γ and transfected into suspension cultured CHO cells for secretory expression of rPoIFN-γ. The rPoIFN-γ was purified by affinity chromatography and identified with SDS-PAGE and Western blotting. Subsequently, the cytotoxicity of rPoIFN-γ was analyzed by CCK-8 test, and the antiviral activity of rPoIFN-γ was evaluated using standard procedures in VSV/PK-15 (virus/cell) test system. Finally the anti-Seneca virus A (SVA) of rPoIFN-γ activity and the induction of interferon-stimulated genes (ISGs) and cytokines were also analyzed. The results showed that rPoIFN-γ could successfully expressed in the supernatant of CHO cells. CCK-8 assays indicated that rPoIFN-γ did not show cytotoxicity on IBRS-2 cells. The biological activity of rPoIFN-γ was 5.59×10⁷ U/mg in VSV/PK-15 system. Moreover, rPoIFN-γ could induced the expression of ISGs and cytokines, and significantly inhibited the replication of SVA. In conclusion, the high activity of rPoIFN-γ was successfully prepared by CHO cells expression system, which showed strong antiviral activity on SVA. This study may facilitate the investigation of rPoIFN-γ function and the development of novel genetically engineered antiviral drugs.
Swine ; Animals ; Cricetinae ; Interferon-gamma/pharmacology* ; Cricetulus ; CHO Cells ; Sincalide ; Recombinant Proteins/pharmacology* ; Antiviral Agents/pharmacology*

The aim of this study was to produce E^rns protein of bovine viral diarrhea virus (BVDV) by using suspensively cultured CHO cells expression system and to analyze the immunogenicity of the purified E^rns protein. In this study, the recombinant eukaryotic expression plasmid pcDNA3.1-BVDV-E^rns was constructed based on the gene sequence of BVDV-1 NADL strain. The E^rns protein was secreted and expressed in cells supernatant after transfecting the recombinant expression plasmid pcDNA3.1-BVDV-E^rns into CHO cells. The expression and purification of the E^rns protein was analyzed by SDS-PAGE, the reactivity was determined with anti-His monoclonal antibodies and BVDV positive serum with Western blotting. Immunogenicity analysis of the E^rns protein was determined after immunizing New Zealand white rabbits, and the serum antibodies were tested by indirect ELISA (iELISA) and indirect immunofluorescence (IFA). The serum neutralizing titer of the immunized rabbits was determined by virus neutralization test. The concentration of the purified E^rns protein was up to 0.886 mg/mL by BCA protein quantification kit. The results showed that the E^rns protein could be detected with anti-His monoclonal antibodies and anti-BVDV sera. Serum antibodies could be detected by iELISA on the 7th day post-prime immunization, and the antibody level was maintained at a high titer until the 28th day post-immunization. The antibody titer was 1:128 000. Furthermore, the expression of the E^rns protein in BVDV-infected MDBK cells could be detected with immunized rabbits sera by IFA. Moreover, antigen-specific neutralizing antibodies of 2.71 log₁₀ was induced in rabbits. In this study, purified BVDV E^rns protein was successfully produced using CHO suspension culture system, and the recombinant protein was proved to have a good immunogenicity, which may facilitate the development of BVD diagnosis method and novel subunit vaccine.
Rabbits ; Animals ; Cricetinae ; Cricetulus ; CHO Cells ; Antibodies, Viral ; Diarrhea Viruses, Bovine Viral/genetics* ; Antibodies, Monoclonal/genetics* ; Diarrhea ; Viral Vaccines/genetics*

Objective:To study the incidences of vitamin K 1 and K 2 deficiency (VKD) in umbilical cord blood (UBC) of neonates and the dynamic changes and influencing factors of serum vitamin K 1 levels after preventive vitamin K 1 supplementation. Methods:From January 2021 to June 2022, neonates born in the Obstetrics Department of our hospital were prospectively enrolled and the levels of vitamin K 1 and K 2 in UBC and serum vitamin K 1 levels at 14 d and 28 d after vitamin K 1 supplementation were measured. The neonates were assigned into hospitalization group and healthy group and further assigned into early-preterm, late-preterm and full-term groups based on gestational age (GA). The incidences of VKD of different GA were studied. Dynamic changes of vitamin K 1 levels were calculated. Multivariate logistic regression was used to analyze the influencing factors of vitamin K 1 levels in hospitalization group at 28 d. Results:A total of 100 neonates were included. 80 neonates were hospitalized, including 25 early-preterm, 25 late-preterm and 30 full-term. 20 were healthy full-term neonates. No significant differences existed in the incidences of VKD of different GA ( P>0.05), however, the overall incidences were high (82.0% and 84.0%, respectively). After preventive vitamin K 1 supplementation, the levels of vitamin K 1 in full-term and preterm groups at 14 d were higher than at birth and 28 d. The levels of vitamin K 1 in hospitalized full-term neonates at 14 d and 28 d were higher than hospitalized preterm neonates. The levels of vitamin K 1 at 28 d in healthy group was significantly higher than hospitalization group ( P<0.05). Multivariate logistic regression analysis showed that maternal complications during pregnancy ( OR=5.889, 95% CI 1.621-21.399, P=0.007) and neonatal antibiotic use ( OR=5.615, 95% CI 1.833-17.221, P=0.003) were risk factors and formula feeding ( OR=0.389, 95% CI 0.193-0.786, P=0.008) was a protective factor for VKD. Conclusions:VKD is common in neonates. The serum vitamin K 1 level increases significantly after preventive vitamin K 1 supplementation. The vitamin K 1 levels of hospitalized full-term neonates at 14 d and 28 d are higher than hospitalized preterm neonates. The levels of vitamin K 1 at 28 d in hospitalized neonates are influenced by feeding methods, maternal complications during pregnancy and neonatal antibiotic use.

Hepatic sinusoidal obstruction syndrome (HSOS) is the most common early complication in patients with hematopoietic stem cell transplantation (HSCT). According to reports, the average incidence of HSOS post-HSCT is 13.7% (0~62%). HSOS may be accompanied by multiple organ failure, and the mortality rate can exceed 80% at 100 days of onset. Defibrotide is currently the safest and most effective drug used to prevent and treat HSOS post-HSCT. It is currently the only drug approved in the United States for the treatment of HSOS with lung/renal dysfunction post-HSCT. In addition, it has been approved in the European Union for the treatment of severe HSOS cases post-HSCT. This article briefly describes and summarizes the research progress, clinical application, pharmacokinetics, efficacy, safety, dose and treatment of defibrotide in HSOS post-HSCT.

Objective:To explore the application of ArcCheck system in the validation of Helical and Direct tomotherapy plans for esophageal cancer and summarize relevant experience.Methods:The Helical and Direct tomotherapy verification plans were established for 32 patients with esophageal cancer at different positions according to the doctor′s instructions, which were verified by the ArcCHECK system to compare the passing rate of the results.The correlation between the volume of the target area and the passing rate of the planned verification was analyzed. The therapeutic verification plan with a small target volume was made. The target area was placed at the center of ArcCHECK phantom and the area of detectors to statistically compare the verification passing rates.Results:Helical plan showed a significantly higher passing rate than the Direct plan ( P<0.01). The correlation coefficients between the target volume and the passing rate of the Helical and Direct plans were -0.364 and -0.042, and the P values were 0.041 and 0.819, respectively. For the Helical plan, when the 3%/2mm criterion was adopted, there was significant difference between placing the high-dose area at the center of the phantom and the area of detectors ( P=0.005), and the passing rate of the latter was higher. There was no significant difference in the other cases (all P>0.05). Conclusions:The passing rate of the Helical plan is generally higher than that of the Direct plan, which may be related to the angular response of the ArcCHECK detector and the fact that more reference points are not included for calculation due to low-dose radiation. In addition, it may also be related to the higher requirements of Direct plan for tomotherapy dose control system. In the Helical verification plan, when the 3%/3mm criterion is adopted, the larger the target volume, the higher the possibility of lower passing rate, whereas the correlation coefficient between them is relatively low. The high-dose area can be verified by the plans at the center of the phantom or the detection point. With the comprehensive consideration, we suggest putting it at the center of the phantom.

ObjectiveTo investigate the value of Model for End-Stage Liver Disease (MELD) and Child-Turcotte-Pugh (CTP) score in predicting the prognosis of patients with hepatic sinusoidal obstruction syndrome (HSOS) associated with Gynura segetum (Lour.) Merr. MethodsA total of 49 patients with HSOS associated with Gynura segetum (Lour.) Merr. who were admitted to Beijing YouAn Hospital, Beijing Ditan Hospital, The Fifth Medical Center of Chinese PLA General Hospital, Tianjin Third Central Hospital, and The First Affiliated Hospital of Xinxiang Medical University from January 2012 to July 2018 were enrolled and followed up for three years, with death as the outcome event. MELD and CTP scores were calculated according to the laboratory examination and clinical data on admission, and according to CTP score, the patients were divided into CTP class A (CTP score 5-6) group(n=8), CTP class B (CTP score 7-9) group(n=23), and CTP class C (CTP score ≥10) group(n=18). The patients were divided into death group(n=12) and survival group(n=37) according to the clinical outcome during follow-up. The Mann-Whitney U test was used for comparison of continuous data between groups, and the Kruskal-Wallis H test was used for ranked data. The area under the receiver operator characteristic (ROC) curve (AUC) was used to investigate the ability of CTP and MELD scores in predicting death. The Kaplan-Meier survival curves were used to determine the long-term prognosis of patients with different CTP and MELD scores, and the log-rank test was used for comparison. The ROC curve was used to evaluate the performance of these two scoring systems in predicting death. ResultsA total of 12 patients died during the 3-year follow-up period. The patients with HSOS had a median MELD score of 13.443 (8.792-18.379), and the death group had a significantly higher MELD score than the survival group ［19.84 (15.49-25.41) vs 11.58 (8.60-15.79), Z=-3.511, P＜0.001］. The patients with HSOS had a CTP score of 6-12, and of all 49 patients, 8 (16.3%) had CTP class A HSOS, 23 (46.9%) had CTP class B HSOS, and 18 (36.7%) had CTP class C HSOS. The mortality rate of the patients increased significantly with the increase in CTP score (χ2=16.078, P＜0.05). The mortality rates of the patients with CTP class A, B, and C HSOS were 0.0%, 13.0%, and 50.0%, respectively (χ2=10343, P＜0.05). The Kaplan-Meier analysis showed that the patients with a MELD score of ＜14.294 4 had a significantly better 3-year prognosis than those with a MELD score of ≥14.294 4 (χ2=14.893, P＜0.001). The higher the CTP score, the poorer the 3-year prognosis of patients (χ2=11.083, P＜0.05). CTP class had an AUC of 0.780 (95% confidence interval ［CI］: 0.639-0.922) in predicting the prognosis of HSOS patients, while MELD score had an AUC of 0.840 (95%CI: 0.722-0.958), and there was no significant difference between the two scores (Z=2.63, P＞0.05). ConclusionBoth MELD and CTP scores can predict the risk of death in patients with HSOS, with similar performance in predicting the prognosis of patients, and further studies are needed to validate their clinical value.

Objective： ：To explore a novel chimeric antigen receptor (CAR)-T cell treatment to treat Multiple Myeloma (MM) via target B cell maturation antigen (BCMA). Methods： ：A CAR-BCMA molecular was constructed based on mouse originated BCMA scFv, and was packaged into lentiviral vector and transfected into T cells from healthy donors to construct CAR-BCMA-T cells. The BCMApositive cell lines A549-BCMA, A549-BCMAOFP and K562-BCMA were constructed as target cells. Then, the CAR-BCMA-T cells were co-incubated with the constructed target cells and human myeloma U266 cells, and the cytotoxic effects of CAR-BCMA-T cells were evaluated via CCK-8 and FACS. Finally, the CAR-BCMA-T cells originated from MM patients were constructed, and its cytotoxicity against A549-BCMA were examined; in addition, the IFN-γ release level in CAR-BCMA-T cells was evaluated by ELISA and FACS. Results: After 11 days’incubation, the CAR-BCMA-T cells originated from healthy donors amplified 300 times with a positive rate of 43%. The BCMApositive target cell lines were constructed successfully. Under an effector : target ratio of 5:1, the killing rates of CARBCMA-T cells against A549-BCMA, K562-BCMA and U266 were about 80%, 60%, and 80%, respectively, which were significantly higher than those against BCMA negative cells; and the cytotoxicity was related to the BCMA expression level in target cells. What’ s more, at the effector : target ratio of 20:1, the CAR-BCMA-T cells originated from MM patients were demonstrated to exhibit a killing rate of more than 95% againstA549-BCMApositive cells, and produced large amount of IFN-γ. Conclusion: CAR-BCMA-T cells originated from both healthy and MM donors were successfully constructed, and they can effectively and specifically kill BCMA positive tumor cells.

Objective To investigate whether GLT25D2 gene regulates autophagy in acetaminophen (APAP)-induced hepatotoxicity injury.Methods GLT25D2+/+ wild-type C57BL/6J mice and GLT25D2-/- C57BL/6J mice were selected as subjects. ①In vivo experiment: 20 for wild-type mice and 20 for GLT25D2-/- mice were respectively divided into phosphate buffer (PBS) control group and APAP intervention group according to random number table, with 10 mice in each group. The hepatotoxicity injury model of mice was reproduced by intraperitoneal injection of 25 g/L APAP solution 500 mg/kg. The PBS control group was intraperitoneally injected with the same amount of PBS. The mice were sacrificed immediately after model reproduction, and the liver tissues were harvested. Western Blot was used to detect the expressions of autophagy-related proteins ATG5, ATG7, microtubule-associated protein 1 light chain 3 (LC3) and P62. The ultrastructural changes in liver tissue were observed under electron microscope to observe the level of autophagy. ②In vitro experiment: primary hepatocytes extracted by two-step collagen perfusion from one GLT25D2+/+wild-type mouse and one GLT25D2-/- mouse were divided into two parts respectively. One part was treated with 5 mmol/L APAP solution. The cells were harvested at 0, 8, and 12 hours, and the expressions of autophagy-related proteins ATG5, ATG7, LC3, and P62 were determined by Western Blot. The other part was transfected with the green fluorescent protein-LC3 plasmid (GFP-LC3) for 24 hours. The cells were cultured with PBS (PBS control group) or 5 mmol/L APAP (APAP intervention group) for 12 hours, and the positive expression of GFP-LC3 was observed under the fluorescence microscope, thereby reflecting the expression of autophagosomes.Results ①In vivo experiment: compared with the corresponding PBS control group, the expressions of the positive-associated proteins ATG5, ATG7 and LC3-Ⅱ in liver tissue of the APAPintervention group were down-regulated in the wild-type and GLT25D2-/- mice, while the expression of the negative correlation protein P62 was up-regulated, indicating that the overall level of autophagy decreased after treatment with APAP. Compared with wild-type mice, the expressions of autophagy positive correlation proteins ATG5 and ATG7 were up-regulated in GLT25D2-/- mice (ATG5/β-actin: 1.21±0.29 vs. 0.84±0.19, ATG7/β-actin:1.29±0.14 vs. 1.54±0.40, bothP > 0.05), LC3-Ⅱ expression was slightly down-regulated (LC3-Ⅱ/β-actin: 0.52±0.06 vs. 0.58±0.06,P > 0.05), while negative correlation protein P62 was down-regulated (P62/β-actin: 1.13±0.94 vs. 1.54±0.40,P > 0.05), indicating that the expression of autophagy in GLT25D2-/- mice was higher than that in wild-type mice. Ultrastructural observation under electron microscope showed that the number of autophagosomes in the liver tissue of wild-type mice did not change significantly after APAP intervention as compared with that in PBS control group, but the number of autophagosomes in GLT25D2-/- mice was increased. ②In vitro experiment: with the prolongation of APAP intervention, the expressions of ATG5 and ATG7 in the primary hepatocytes of wild-type and GLT25D2-/-mice were up-regulated, LC3 was slightly fluctuated, and the expression of negative-related protein P62 was gradually down-regulated. The peak value or the trough value reached at 12 hours. It was indicated that the expression of autophagy in APAP-stimulated cells was enhanced with a time-dependent manner. Compared with wild-type mice, the expressions of autophagy correlation proteins ATG5, ATG7, LC3-Ⅱ and P62 were up-regulated in GLT25D2-/- mice at 12 hours (ATG5/β-actin: 0.93±0.09 vs. 0.74±0.06, ATG7/β-actin: 0.80±0.09 vs. 0.65±0.10, LC3-Ⅱ/β-actin:1.35±0.30 vs. 1.15±0.20, P62/β-actin: 0.36±0.02 vs. 0.31±0.03, allP > 0.05), indicating that the expression of autophagy was enhanced after gene knockout. Fluorescence microscopy showed that GFP-LC3 positive cells in both wild-type and GLT25D2-/- mice hepatocytes were significantly increased after APAP intervention as compared with those of PBS control group, and the proportion of GFP-LC3 positive cells in GLT25D2-/- mice was significantly higher than that in wild-type mice (0.64±0.08 vs. 0.36±0.05,P < 0.05).Conclusions GLT25D2 is a negative regulator of autophagy. Knockout of GLT25D2 gene can enhance the autophagy level of APAP-induced hepatotoxicity injury in mice.