Objective To explore the correlation between serum vitamin D(VD3)level differences and immune inflammatory markers in elderly sepsis patients.Methods A total of 103 elderly patients with sepsis(aged 65-99 years)in the ICU of the First Affiliated Hospital of Kunming Medical University from January 2020 to December 2022 were collected and divided into two groups according to the diagnostic criteria for VD3 deficiency:VD3 deficiency group(n=32)and VD3 severe deficiency group(n=71).Correlation analysis was conducted by comparing the differences in serum 25-(OH)-D3(VD3)levels,immune function-related indicators upon admission(blood routine,infection-related proteins,combined detection of 12 cytokines,absolute count analysis of lymphocytes and subgroups,quantitative determination of infection-related immune cells,immunoglobulin,and complement),illness severity,and prognostic indicators(APACHE-II score,SOFA score,duration of ICU stay,and 28-day mortality rate).Result(1)Serum VD3 levels were lower in elderly patients with sepsis.No patient was in the VD3 normal or insufficient group.Patients with severe VD3 deficiency had higher APACHE-II scores,SOFA scores,and 28-day mortality rates than those with VD3 deficiency,and these scores were negatively correlated with serum VD3 levels(P<0.001),while the difference in ICU stay duration between the two groups was not statistically significant(P>0.05);(2)WBC,PCT,CRP,and CD4/CD8 in the VD3 deficiency group were all lower than those in the VD3 severe deficiency group(P<0.05),while IL-6,IL-10,CD45+,CD3+/CD45+,and CD19+Abs were all higher than those in the VD3 severe deficiency group(P<0.05);In the VD3 deficiency group,VD3 levels were positively correlated with CD45+(P<0.05 for all),while negatively correlated with IL-6,IL-10,PCT,and CRP(P<0.05 for all);In the VD3 severe deficiency group,there were fewer corre-lation indicators and the correlation strength was not as strong as that in the VD3 deficiency group.Conclusion(1)Elderly patients with sepsis generally have lower levels of VD3,with lower levels associated with more severe illness and poorer prognosis;(2)In elderly sepsis patients,compared to patients with severe VD3 deficiency,patients with VD3 deficiency have lower levels of inflammation,stronger cellular immune response,and stronger correlation,suggesting that the effects of different VD3 levels on immune inflammatory responses may vary in elderly sepsis patients.

Sepsis is a syndrome caused by a dysregulated host response to infection that leads to life-threatening organ dysfunction.The immune response in sepsis is usually divided into two stages:hyperimmunity and immunosuppression.Immunosuppression is considered as the main cause of secondary infection and increased mortality in patients.This article reviews the main mechanisms of immunosuppression in sepsis,including dysfunction of immune cells,release of immunosuppressive cytokines,excessive activation of immunomodulatory cells,increased expression of immune checkpoints,and epigenetic dysregulation that have attracted much attention in recent years.At the same time,it also summarizes the existing and some new immunotherapy strategies.It includes immunostimulatory cytokines(such as GM-CSF,IL-7,IL-15),immune checkpoint inhibitors(such as PD-1/PD-L1 antibody,CTLA-4 antibody,TIM-3 antibody)and emerging immunotherapy methods(such as mesenchymal stem cells,calprotectin inhibitors,triggering receptor expressed on myeloid cells 1 inhibitors).This article focuses on epigenetic regulation mechanisms and emerging immunotherapies such as mesenchymal stem cells,aiming to provide theoretical support for individualized precision immunotherapy in patients with sepsis.

OBJECTIVE To investigate the intervention effect of Dabufei decoction on acute lung injury in rats with high altitude hypoxia by regulating the expression of the HIF-1α/NLRP3 signaling pathway and related molecules. METHODS Sixty SPF SD rats were randomly divided into blank group, model group, positive drug group, Dabufei decoction high-dose, medium-dose, and low-dose groups with 10 rats in each group. After 3 d of adaptation to feeding, the rats in the blank group and model group were given the same amount of normal saline by gavage, and the rats in Dabufei decoction high-dose, medium-dose, and low-dose groups were given gavage for 14 d, respectively. The positive drug group was given dexamethasone by intraperitoneal injection for three consecutive days before entering the chamber. Except for the blank group, the rats in each group were exposed to hypoxia in the experimental animal low-pressure simulation chamber from the 15th day for three consecutive days. At the end of the experiment, the wet to dry ratio(W/D) of the rat lung tissue was detected. The morphology of lung tissue was observed by HE staining. ELISA detected the levels of IL-1β and IL-18 in serum. Western blotting and RT-qPCR were used to detect the protein and mRNA expressions of HIF-1α, NLRP3, GSDMD, and caspase-1 in the lung tissue of rats. RESULTS The W/D value showed that compared with the blank group, the W/D of the model group was significantly increased(P<0.01). Compared with the model group, the W/D of rats in the positive drug group, Dabufei decoction high-dose group, medium-dose, and low-dose groups was significantly decreased(P<0.01 or P<0.05). HE results showed that compared with the blank group, alveolar septum thickening, pulmonary interstitial congestion, edema, inflammatory cell infiltration, and a small amount of exudation in the alveolar cavity were seen in the lung tissue of the model group. Compared with the model group, the thickening of alveolar walls in the positive drug group, Dabufei decoction high-dose group, medium-dose, and low-dose groups were reduced, and the pulmonary interstitial congestion, edema, and inflammatory cell infiltration were significantly reduced. ELISA results showed that IL-1β and IL-18 in rat serum were significantly higher in the model group than in the blank group(P<0.01). Compared with the model group, the levels of IL-1β and IL-18 in the serum of the rats in the positive drug group, Dabufei decoction high-dose group, medium-dose, and low-dose groups were significantly decreased(P<0.05 or P<0.01). Moreover, the results of Western blotting and RT-qPCR showed that compared with the blank group, the relative protein and mRNA expressions of HIF-1α, NLRP3, GSDMD, and caspase-1 in the lung tissue of the model group were significantly increased(P<0.01). Compared with the model group, the relative protein and mRNA of HIF-1α, NLRP3, caspase-1 and GSDMD in the positive drug group and Dabufei decoction high-dose group were significantly decreased(P<0.05 or P<0.01), the relative protein of HIF-1α, NLRP3, caspase-1 and GSDMD in Dabufei decoction medium-dose group were significantly decreased and HIF-1α, caspase-1 mRNA were significantly decreased(P<0.05), the relative protein of HIF-1α and GSDMD in the low-dose group was decreased(P<0.05). The positive drug group and Dabufei decoction high-dose group had the more significant effect. CONCLUSION Dabufei decoction can regulate the HIF-1α/NLRP3 signaling pathway, inhibit pyroptosis and reduce inflammation, and has a certain protective effect on acute lung injury in rats with high altitude hypoxia.

This study was performed to screen the sequence of specific nanobody targeting SARS-CoV-2 S protein,and to express and purify recombinant anti-SARS-CoV-2 nanobodies,and evaluate their characteristics and application potential.Based on the previously constructed natural bacteriophage nanobody display library,the anti-SARS-CoV-2 S protein nanobody sequences were screened,with biotinylated SARS-CoV-2 S protein receptor-binding domain antigen as target,by biotin-streptavidin liquid phase screening method,phage-ELISA and sequencing.Recombinant anti-SARS-CoV-2 S protein nanobody expression vectors were constructed,the protein was induced by IPTG and then purified,and their characteristics and application potential were analyzed by SDS-PAGE,Westem blot and ELISA methods.Two anti-SARS-CoV-2 S protein nanobody sequences were successfully obtained,and the corresponding prokaryotic expression bacteria were constructed.The recombinant anti-SARS-CoV-2 S protein nanobody was expressed and purified successfully,which can specifically bind to SARS-CoV-2 S protein.In conclusion,the recombinant anti-SARS-CoV-2 S protein nanobodies have screened and expressed successfully,which provides a basis for its application in SARS-CoV-2 detection and treatment,and facilitates related researches.

ObjectiveTo observe the effect of Huangqi Baihe granules on the hypoxia-inducible factor 1α （HIF-1α）/nuclear factor-κB （NF-κB）/NOD-like receptor hot protein domain related protein 3 （NLRP3） signaling pathway in a rat model of high altitude hypoxia. MethodSixty male SPF SD rats were randomly divided into blank group， model group， dexamethasone group （5 mg·kg^-1）， and high， middle， and low-dose groups of Huangqi Baihe granules （4.1， 2.05， 1.025 g·kg^-1）. Among them， each Chinese medicine group was administrated orally for continuously 14 d， once a day， and the dexamethasone group was injected intraperitoneally for continuously 3 d as the positive control group. On the 15^th d， the model group， dexamethasone group， and high， middle， and low dose groups of Huangqi Baihe granules were exposed to the simulated high altitude， low pressure， and low oxygen environment in the animal low-pressure simulation cabin， and the exposure lasted for 3 d. Blood was collected from the abdominal aorta and serum was separated， and the brain tissue was taken after being killed. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in brain tissue. Enzyme-linked immunosorbent assay （ELISA） was used to detect the content of tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and interleukin-1β （IL-1β） in rat serum. Western blot was used to detect HIF-1α， NLRP3， phosphorylated nuclear factor-κB （p-NF-κB）， NF-κB， desquamation D （GSDMD）， and cysteine aspartate-specitis protein-1（Caspase-1） in rats of each group. The mRNA expression levels of HIF-1α， NLRP3， NF-κB p65， GSDMD， and Caspase-1 were detected by real-time quantitative polymerase chain reaction （Real-time PCR）. ResultThe results of HE staining showed that as compared with the normal group， the pathological sections of brain tissues in the model group showed that pyramidal cells were loosely arranged and distributed in disorder， with different sizes. Compared with the model group， the pathological changes in pyramidal cells in the dexamethasone group and high and middle-dose groups of Huangqi Baihe granules were reduced. The results of ELISA showed that as compared with the normal group， the content of TNF-α， IL-6， and IL-1β in the serum of rats in the model group was significantly higher （P<0.01）. Compared with the model group， the content of TNF-α， IL-6， and IL-1β in the serum of rats in the dexamethasone group and high and middle-dose groups of Huangqi Baihe granules decreased significantly （P<0.05， P<0.01）. The results of Western blot showed that as compared with the normal group， the relative protein expression levels of HIF-1α， NLRP3， p-NF-κB p65， GSDMD， and Caspase-1 in the brain tissue of the model group were significantly higher （P<0.01）. As compared with the model group， the relative expressions of HIF-1α， NLRP3， p-NF-κB p65， GSDMD， and Caspase-1 in the brain tissue of rats in the dexamethasone group and the high-dose group of Huangqi Baihe granules were significantly decreased （P<0.05， P<0.01）. The relative protein expression levels of HIF-1α， NLRP3， p-NF-κB p65， and Caspase-1 in the brain tissue of rats in the middle-dose group of Huangqi Baihe granules decreased significantly （P<0.01）， and the relative protein expression of HIF-1α in the brain tissue of rats in the low-dose group of Huangqi Baihe granules was reduced （P<0.05）. The Real-time PCR analysis showed that as compared with the normal group， the mRNA expression levels of HIF-1α， NLRP3， NF-κB p65， GSDMD， and Caspase-1 in the brain tissue of the model group were significantly increased （P<0.01）. As compared with the model group， the mRNA expression levels of HIF-1α， NLRP3， NF-κB p65， GSDMD， and Caspase-1 in the brain tissue of rats in the dexamethasone group were significantly decreased （P<0.01）. The mRNA expression levels of HIF-1α， NF-κB p65， GSDMD， and Caspase-1 in the brain tissue of rats in the high-dose group of Huangqi Baihe granules decreased significantly （P<0.01）. The mRNA expression levels of HIF-1α， NLRP3， and Caspase-1in the brain tissue of rats in the middle-dose group of Huangqi granules decreased （P<0.05， P<0.01）. ConclusionThe protective effect of Huangqi Baihe granules on acute brain injury in low-pressure hypoxic rats may be related to the HIF-1α/NF-κB/NLRP3 signaling pathway.

To study the methods and principles for simultaneous treatment in the children with pectus excavatum (PE) combined with congenital cardiothoracic diseases.
 Methods: The medical records of all children, who underwent simultaneous repair of PE combined with congenital cardiothoracic diseases, were retrospectively reviewed in Hunan Children's Hospital from January 2007 to September 2018. The patients were divided into a PE combined with congenital heart disease (CHD) group (n=17) and a PE combined with thoracic disease group (n=10). The repair with a custom-made sternal lifting device, a Nuss repair, were performed in the treatment of PE, and the correction of the CHD was performed by heart open surgery using cardiopulmonary bypass (through sternotomy or right infra-axillary thoracotomy) or by transcatheter closure under echocardiography or X-ray-guided percutaneous intervention in the PE combined with CHD group. The children in the PE combined with thoracic disease group underwent thoracic surgery plus Nuss procedure concurrently.
 Results: All 27 pediatric patients underwent simultaneous repair of the PE combined with congenital cardiothoracic diseases. In the PE combined with CHD group, the duration of hospital stay ranged from 8.0 to 25.0 (13.2±4.8) days. Two patients had delayed healing of the surgical wound and 1 patient developed a small left pleural effusion postoperatively. In the PE combined with thoracic disease group, the duration of hospital stay ranged from 10.0 to 34.0 (19.9±7.5) days. One patient was complicated with chylothorax and 2 patients were complicated with pleural effusionin. The treatment for the patients in the 2 groups was satisfactory. No severe complications like surgical death, severe bleeding, chest organ injuries, and implant rejections were observed.
 Conclusion: According to the characteristics of patients, individualized programs should be selected in order to correct children's PE combined with congenital cardiothoracic diseases in the same period, which are safe, effective and can avoid the risk of multiple operations and anesthesia, and can reduce the financial burden of family.
Cardiac Surgical Procedures ; Child ; Funnel Chest ; surgery ; Heart Defects, Congenital ; Humans ; Minimally Invasive Surgical Procedures ; Retrospective Studies ; Sternotomy ; Treatment Outcome

Objective To screen the optimized Buyang Huanwu Decoction (BYHWD);To verify it. Methods H2O2 was used to induce PC12 cell oxidative stress models. MTT method was used to determine the prevention effects of BYHWD at different concentrations (0.1, 0.2, 0.5, 1.0, 2.0, 3.5 mg/mL) on in vitro oxidative stress cell models to define the optimized concentration. Orthogonal design was used to divide BYHWD single medicine into decomposed BYHWD groups, control group (only with DMEM), normal group (without H2O2 and medicine processing), and model group, to investigate the protective effects on PC12 cells. Optimized BYHWD was screened to decide the compatibility ratio of each medicine. MTT was used to detect the cell survival rate in each group. Middle cerebral artery occlusion was used to replicate MACO rat models. SD rats were randomly divided into sham-operation group, model group, BYHWD group and optimized BYHWD high-, medium-and low-dose groups. Each medication group was given relevant medicine for gavage. The screened results were verified. Results Compared with other decomposed BYHWD groups, the protective effects of the compatibility of Astragali Radix+Chuanxiong Rhizoma+Pheretima on PC12 cells was the best (P<0.05), which was nearly equaled to BYHWD. Compared with the model group, BYHWD and the optimized one could evidently reduce cerebral cortex infarction area and improve the impaired brain edema (P<0.05), and the medium-dose group was the best. Conclusion The optimized BYHWD ratio is:Astragali Radix:Chuanxiong Rhizoma:Pheretima=10:3:1.

Objective To investigate the effects of L-carnitine on myocardial enzymes in infants with congenital heart disease undergoing open cardiac operation with cardio pulmonary bypass (CPB).Methods From Mar.2011 to Nov.2012,there were 60 infants with ventricular septal defects,whom were divided randomly into the test (n =30) and control (n =30) groups.L-carnitine was put in the cold crystal cardiac arresting liquid in the test group (6 g/L),other experimental conditions were the same between two groups.Before CPB,at 1 h,24 h,7 d after the pump-off,venous blood was drawn to test the level of serum creatine kinase (CK),CK-MB isozyme,cardiac Troponin I (cTnI).Blood samples were taken at different time points for the analysis of tlymphocyte and its subtype by flow cytometry.Observing the dopamine and other vasoactive drug 8 hours and 24 hours after the pump-off,using data obtained from the t-test done statistically.Results The levels of CK and CK-MB and cTnI had no difference between two groups before operation.From the end of CPB,the levels of CK and CK-MB and cTnI were significantly lower in the test group than that in the control group (P ＜0.05).The automatic jump higher rate (95％ vs 70％,P ＜ 0.05) and the amount of dopamine were significantly less than the control group (P ＜ 0.05).Conclusions L-carnitine improved obviously microcirculation of ischemic cardiac reperfusion injury undergoing open cardiac operation under CPB and heart function.It had a good protective effect on myocardium in infants.

OBJECTIVETo establish the double HPLC fingerprints of water-soluble composition and amino acids precolumn derivative reagent of 13 batches of Isatidis Radix micropower.

METHODThe gradient elution was adopted with Hypersil BDS C18 column (4.6 mm x 250 mm, 5 microm). Water-soluble ingredients were detected with acetonitrile-0.1% phosphoric acid solution as mobile phase, flow rate 0.5 mL x min(-1), column temperature 20 degrees C, and the injection volume 10 microL. Amino acid ingredient were derived by PITC, and then were detected with mobile phase of 0.1 mol x L(-1) sodium acetate buffer solution (pH 6.5) - acetonitrile, flow rate 1.0 mL x min(-1), column temperature 30 degrees C, and the injection volume 5 microL. Both of the absorption wavelengths were 254 nm. Pharmacopoeia Commission "Chinese chromatographic fingerprint evaluation system (version 2.0)" was adopted to analyse, fingerprints of Isatidis Radix micropower were established, at the same time 4 main ingredients were recognized by the SPSS cluster analysis.

RESULTCommon mode of Fingerprint to water-soluble and amino acids ingredient of Isatidis Radix micropower was established, then adenosine, epigoitrin and 15 amino acids were identified as characteristic peaks. Cluster analysis showed that different kinds of the herbal Isatidis Radix micropower from different areas were different levels in the main ingredients.

CONCLUSIONDouble fingerprints of Isatidis Radix micropower is established. Each peak is optimally separated in chromatogram, which provides a scientific basis for quality control of Isatidis Radix micropower.

Objective To establish a dose-effect curve of premature condensation chromosome ring (PCC-R)in lymphocytes of human peripheral blood after exposed to high doses of Y-rays. Methods Peripheral blood samples was drawn from three healthy individuals, and exposed to 60Co γ-rays with doses between 0 and 30 Gy. The frequencies of PCC-R in premature condensation chromosome (PCC) cells obtained by Okadaic acid (OA) induction were calculated, and a dose-effect curve was fitted. Results PCC index tapered with dose. Frequencies of PCC-R per cell increased until 20 Gy, and then saturation was observed. The results were fitted to a lineal model up to 20 Gy: y = - 0.020 + 0.052D ,where y was the frequencies of PCC-R per cell, D was the radiation dose (Gy). Conclusins The highest dose could be estimated is 20 Gy by the dose-effect curve established with PCC-R method. Its utility and validity will be verified in the future application of radiation accident.