Objective To observe clinical efficacy of probiotics agents in the prevention and treatment of severe pulmonary infection in elderly patients with antibiotic-associated diarrhea (AAD).Methods 60 cases of elderly patients with severe pulmonary infection (more than or equal to 60 years old) were randomly divided into the treatment group ( 31 cases ) and control group ( 29 cases ) , the control group received broad-spectrum antibiotics or using two linked above anti-infection treatment, the treatment group were added with probiotic agent ( lactobacillus complex capsules).The diarrhea, the use of antibiotics and the stool routine, bacteria before and after 5, 10, 15 days of group were observed.Results After 15 days treatment, the number of Escherichia coli in treatment group was lower than that in control group (P<0.05), the number of Bacterium lacticum and Bifidobacterium bifidum in treatment group were higher than those in control group ( P<0.05 ) .The AAD rate in treatment goup was 12.90%, which was lower than 41.38% in control group (P<0.05).There were significant differences in beginning time and duration of diarrhea between two groups (P<0.05). Conclusion The intestinal probiotics reduced induced by antibiotics in elderly patients with severe pulmonary infection , the probiotics agents could redress intestinal flora imbalance, keep the steady state of intestinal flora, and prevent and cure the antibiotic-associated diarrhea.

An efficient method for the isolation and purification of 12,13-dihydroxyeuparin from Radix Eupatorii Chinensis by high speed counter-current chromatography (HSCCC) was established in this paper.The ether extracts of Radix Eupatorii Chinensis were purified by HSCCC with a solvent system of hexyl hydride ethyl acetate-methanol-water (1∶2∶1∶2,v/v/v/v).The upper phase was used as the stationary phase and the lower phase as the mobile phase.About 8.4 mg of 12,13-dihydroxyeuparin was obtained from 200 mg of ether extracts from Radix Eupatorii Chinensis in one-step HSCCC separation,with the purity of 96.71％,as determined by HPLC.After methanolwater recrystallization,the purity of 12,13-dihydroxyeuparin reached 99.83％.Such a simple and effective method was fairly useful to prepare pure compound as reference substances for related study on Radix Eupatorii Chinensis.

A reversed phase high performance liquid chromatography （HPLC） method was established for the simultaneous determination of 12, 13-dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture. A Grace Apollo Cl8 column （250 mm × 4.6 mm, 5 μm） was used as the stationary phase and the mobile phase was composed of acetonitrile and aqueous phosphoric acid （0.2%, v/v）. Gradient elution was carried out at the flow rate of 1.0 mL/min and the column temperature was 30 ℃. An ultraviolet （UV） detector was used with a selected wavelength of 240 nm. Calibration curves were linear within the concentration range of 4.6-45.75 μg/mL for 12, 13-dihydroxyeuparin （r〉0.9999） and 106.9-1068.9μg/mL for glycyrrhizic acid （r〉0.9999）, respectively. Recoveries were 102.18% for 12, 13-dihydroxyeuparin and 101.17% for glycyrrhizic acid. The method developed could be applied to the simultaneous determination of 12, 13- dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture.

A reversed phase high performance liquid chromatography (HPLC) method was established for the simultaneous determination of 12,13-dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture.A Grace Apollo C18 column (250 mm× 4.6 mm,5 μm) was used as the stationary phase and the mobile phase was composed of acetonitrile and aqueous phosphoric acid (0.2％,v/v).Gradient elution was carried out at the flow rate of 1.0 mL/min and the column temperature was 30 ℃.An ultraviolet (UV) detector was used with a selected wavelength of 240 nm.Calibration curves were linear within the concentration range of 4.6-45.75 μg/mL for 12,13-dihydroxyeuparin (r＞0.9999) and 106.9-1068.9μg/mL for glycyrrhizic acid (r＞0.9999),respectively.Recoveries were 102.18 ％ for 12,13-dihydroxyeuparin and 101.17％ for glycyrrhizic acid.The method developed could be applied to the simultaneous determination of 12,13-dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture.

A high-speed counter-current chromatography (HSCCC) method was successfully developed for the preparative separation and purification of deoxyschizandrin from Schisandrae Sphenantherae Fructus in one step. The purity of deoxyschizandrin was 98.5%, and the structure was identified by MS, UV and NMR. This method was simple, fast, convenient and appropriate to prepare pure compound as reference substances for related research on Schisandrae Sphenantherae Fructus.

Objective To investigate the effects of B lymphocyte-induced maturation protein-1 ( Blimp-1) on the number and function of splenic lymphocytes.Methods The mice with defective Blimp-1 in T cells were generated by cross-breeding B6.Blimp-1flox/flox mice with B6.Lck-Cre mice.The mononuclear lymphocytes isolated from spleen of T cell conditional Blimp-1 knockout (Blimp-1CKO) mice and wild type ( WT) C57/B6 mice were comparatively analyzed.Alterations of CD4+T and CD8+T cell subsets, the secre-tion of cytokines as well as the expression of C-C chemokine receptor type 7 ( CCR7 ) and Sphingosine-1-phosphate receptor 1 (S1P1) in mice from the two groups were analyzed by flow cytometry.The changes of CD19+B cell subsets were also detected.Results Compared with WT mice, the total numbers of mononu-clear cells, T and B lymphocytes were all significantly increased in Blimp-1CKO mice ( P<0.05) .The ab-solute numbers of CD4+T, CD8+T and CD19+CD5+CD1d+B cells in mice form Blimp-1CKO group were higher than those of the control group (P<0.05), however, no significant differences with the percentages of these cell populations were observed between two groups.Higher numbers and percentages of CD19+CD5+B cells were detected in mice from Blimp-1CKO group (P<0.01).The Blimp-1CKO mice showed increased secretion of IFN-γ, TNF-α, IL-17 and IL-2, but decreased expression of CCR7 on CD8+T cells as com-pared with WT mice (P<0.05).No significant differences with the changes of S1P1 were found between the two groups.Conclusion Blimp-1 played an important role in the maintenance of number, phenotype and function of T cells.Furthermore, not only T cells but also B cell subsets in mice were affected by the dele-tion of Blimp-1 in T cells.

Objective:To investigate the clinical efficacy of gastroscopic 'sandwich’ injection and sequential laparoscopic splenectomy combined with pericardial devascularization in the treatment of gastric variceal bleeding.Methods:The retrospective cohort study was conducted. The clinical data of 52 patients with cirrhotic portal hypertension combined with gastric variceal bleeding who were admitted to Affiliated Hospital of Jiaxing University between March 2011 and October 2019 were collected. There were 33 males and 19 females, aged (63±9)years, with a range of 41-83 years. Of the 52 patients, 31 undergoing gastroscopic 'sandwich’ injection and sequential laparoscopic splenectomy combined with pericardial devascularization and 21 undergoing laparoscopic splenectomy combined with pericardial devascularization were allocated into sequential group and laparoscopic group, respectively. Observation indicators: (1) surgical situations; (2) complications; (3) changes in gastric varices after treatment; (4) follow-up. Follow-up was performed using telephone interview combined with outpatient examination to detect survival of patients up to December 2019. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was analyzed by the t test. Measurement data with skewed distribution were represented as M (range), and comparison between groups was analyzed by the Mann-Whitney U test. Count data were expressed as absolute numbers, and comparison between groups was analyzed using the chi-square test or Fisher exact probability. Ordinal data was analyzed by nonparametric rank sum test. Results:(1) Surgical situations: patients of sequential group and laparoscopic group underwent surgery successfully. The operation time and volume of intraoperative blood loss of the sequential group were (112±16)minutes and (57±11)mL, respectively, versus (103±14)minutes and (55±9)mL of the laparoscopic group, showing no significant difference in the above indicators between the two groups ( t=0.963, 1.052, P>0.05). (2) Complications: 11 patients in the sequential group had postoperative complications including 1 case with perioperative bleeding, 2 cases with postoperative gastrointestinal rebleeding, 4 cases with ascites, 4 cases with portal vein thrombosis. There was no death in the sequential group. Sixteen patients in the laparoscopic group had postoperative complications including 2 cases with perioperative bleeding, 6 cases with postoperative gastrointestinal rebleeding, 4 cases with ascites, 4 cases with portal vein thrombosis. Three patients died in the laparoscopic group. There was no significant difference in the cases with perioperative bleeding or cases with ascites between the two groups ( P>0.05) and no significant difference in the cases with portal vein thrombosis or death between the two groups ( χ2=0.082, 0.082, P>0.05). There was a significant difference in the cases with postoperative gastrointestinal rebleeding between the two groups ( P<0.05). Cases with postoperative gastrointestinal rebleeding, cases with ascites, cases with portal vein thrombosis in the sequential group were cured after the treatment of gastroscopy, low salt diet combined with diuretic or low dose warfarin, respectively. Of the 6 patients with postoperative gastrointestinal rebleeding in the laparoscopic group, 3 were cured after the treatment of gastroscopy and 3 died due to failure to rescue in time. Cases with ascites and cases with portal vein thrombosis in the laparoscopic group were cured after the treatment of low salt diet plus diuretic or low dose warfarin, respectively. (3) Changes in gastric varices after treatment: at postoperative 6 months, 31 patients in the sequential group were diagnosed with negative gastric varices; 15 of 21 patients in the laparoscopic group were diagnosed with negative gastric varices, 3 cases were diagnosed with obvious gastric varices and 3 cases were diagnosed with severe gastric varices. There was a significant difference in the cases with gastric varices between the two groups ( Z=-3.128, P<0.05). At postoperative 12 months, 29 patients in the sequential group and 13 patients in the laparoscopic group were diagnosed with negative gastric variceal. There were 2 patients in the sequential group diagnosed with obvious gastric varices, and 8 patients in the laparoscopic group diagnosed with gastric varices including 3 cases with obvious gastric varices and 5 cases with severe gastric varices. There was a significant difference in the cases with gastric varices between the two groups ( Z=-2.933, P<0.05). Cases with obvious gastric varices in the sequential group were cured after the treatment of gastroscopy. Cases with obvious or severe gastric varices in the laparoscopic group were cured after the treatment of gastroscopy except 1 died due to massive gastrointestinal hemorrhage. (4) Follow-up: 52 patients were followed up for 1-8 years, with a median time of 4 years. All the 31 patients in the sequential group and 18 ptients in the laparoscopic group survived. Conclusion:Gastroscopic 'sandwich’ injection and sequential laparoscopic splenectomy combined with pericardial devascularization in the treatment of gastric variceal bleeding has low recurrence rate of varicosity and low incidence of rebleeding.

Objective To explore the characteristics of TCM syndromes in patients with coronary heart disease complicated with hypertension and diabetes based on latent structure model.Methods A multi-center cross-sectional study was used to collect the four diagnostic information of patients with coronary heart disease by using the Four Diagnostic Collection Scales of TCM developed by our research group,and a clinical database was established.Combined with frequency analysis and expert opinions,the four diagnostic information was screened to construct a latent structure model.Results Totally 702 patients with coronary heart disease complicated with hypertension and diabetes were collected.A total of 40 high-frequency four-diagnosis symptoms were screened,and 18 latent variables were obtained by latent structure model analysis.The model BIC score was-17 604.38.The latent variables in the model were comprehensively clustered,and it was found that the syndrome elements of disease location in patients with coronary heart disease complicated with hypertension and diabetes were mainly heart,liver,kidney,etc.,and the syndrome elements of disease nature were mainly qi deficiency,yin deficiency,qi stagnation,phlegm turbidity,blood stasis,etc.The syndrome types were mainly phlegm and blood stasis syndrome,spleen and kidney yang deficiency syndrome,liver depression and qi stagnation syndrome,liver and kidney yin deficiency syndrome,damp-heat intrinsic syndrome,and the mutual information and information coverage between each syndrome type and the four diagnostic information were suggested.Conclusion Latent structure model analysis can provide a qualitative and quantitative basis for the syndrome classification of patients with coronary heart disease complicated with hypertension and diabetes.

Objective:We observed and compared the differences in immune reconstruction between single-infusion anti-B-cell maturation antigen (BCMA) , chimeric antigen receptor T cells (CAR-T) , and combined infusion of anti-CD19 CAR-T cells in the treatment of recurrent/refractory multiple myeloma (RRMM) .Methods:Sixty-one patients with RRMM who underwent CAR-T cell therapy in our hospital from June 2017 to December 2020 were selected. Among them, 26 patients received anti-BCMA target, and 35 patients received anti-BCMA combined with anti-CD19 target. Using flow cytometry, we determined T cell subsets (CD3 +, CD4 +, CD8 +, CD4 +/CD8 +) , B cells (CD19 +) , and NK cells (CD16 + CD56 +) at different time points before and after CAR-T treatment, and detected immunoglobulin IgG, IgA and IgM levels by immunoturbidimetry. We compared the reconstruction rules of lymphocyte subsets and immunoglobulins in the two groups. Results:CD8 + T lymphocytes recovered most rapidly after the infusion of CAR-T cells, returning to pre-infusion levels at 3 months and 1 month after infusion, respectively[BCMA: 695 (357, 1264) /μl vs 424 (280, 646) /μl; BCMA+CD19: 546 (279, 1672) /μl vs 314 (214, 466) /μl]. NK cells returned to normal levels at 3 months after infusion in both groups[BCMA: 171 (120, 244) /μl, BCMA+CD19: 153 (101, 218) /μl (Normal reference range 150-1100/μl) ]; however, the NK cells were not maintained at stable levels in the BCMA CAR-T cells group. The recovery of CD4 + T lymphocytes in both groups was slow and remained persistently low within 12 months after infusion, and no recovery was observed in most patients. The reversal of the ratio of CD4 +/CD8 + lasted for more than a year. The levels of CD19 + B cells in both groups returned to baseline 3 months after infusion[BCMA: 62 (10, 72) /μl vs 57 (24, 78) /μl; BCMA+CD19: 40 (4, 94) /μl vs 29 (14, 46) /μl]. IgG returned to the pre-infusion level 12 months after infusion in the group with anti-BCMA cells alone, but not in the group with combined infusion of CD19 CAR T cells[7.82 (6.03, 9.64) g/L vs 6.92 (4.62, 12.76) g/L]. IgA returned to pre-infusion levels at 9 and 12 months after infusion, respectively[BCMA: 0.46 (0.07, 0.51) g/L vs 0.22 (0.12, 4.01) g/L; BCMA+CD19: 0.46 (0.22, 0.98) g/L vs 0.27 (0.10, 0.53) g/L]. IgM in both groups returned to pre-infusion levels 6 months after infusion[BCMA: 0.43 (0.06, 0.60) g/L vs 0.20 (0.13, 0.37) g/L; BCMA+CD19: 0.53 (0.10, 0.80) g/L vs 0.16 (0.11, 0.28) g/L]. There was no significant difference in the indexes of lymphocyte subpopulation reconstruction and immunoglobulin recovery between the two groups at each time point. Conclusion:This study showed that in patients with RRMM treated with CAR-T cells, the appropriate target antigen can be selected without considering the difference of immune reconstruction between anti-BCMA CAR-T and combined anti-CD19 CAR-T therapy.

Biological hydroxyapatite(BHA)is widely used in the treatment of clinical bone defects due to its good biocompatibility and osteoconductivity.The clinical application of mateiral size is based on the principle of bone defect area adaptation,which contributes to diversity of BHA sizes.However,different sizes correspond to different hierarchi-cal levels of bone biomimicry.As the size changes,the bone biomimicry hierarchy evolves accordingly and influences the process of bone repair and regeneration through osteo-coagulo-immunomodulation,leading to unstable bone graft outcomes.Therefore,this paper reviews the size effect of clinical BHA,analyzes the multilevel structure of natural bone,proposes the evolution of bone biomimetic hierarchy triggered by the size of BHA,and further analyzes the size-media-ted osteo-coagulo-immunomodulation.Based on the hierarchical levels of bone and its osteo-coagulo-immunomodula-tion effect,we provide a new understanding of the biolog-ical principle of the size effect of biomaterials and a theo-retical basis for the basic research and clinical application of different size BHA materials.