ObjectiveTo observe the effects of Youguiwan on bone metabolism and bone morphogenetic protein-2 （BMP-2）/Smad signaling pathway in ovaries-removed rats with osteoporosis and study the mechanism of Youguiwan in the prevention and treatment of osteoporosis. MethodA postmenopausal rat model of osteoporosis was prepared by bilateral ovariectomy. The 40 female SD rats were randomly divided into five groups， including sham operation group， model group， alendronate sodium group （0.1 mg·kg^-1）， and high-dose and low-dose （5.36 and 2.68 g·kg^-1） groups of Youguiwan. The drug was given seven days after modeling， once a day for 12 weeks. After the treatment， the changes in femur tissue structure were observed by micro-CT， including bone mineral density （BMD）， bone volume/total volume （BV/TV）， trabecular number （Tb.N）， trabecular thickness （Tb.Th）， bone surface/bone volume （BS/BV）， and trabecular separation （Tb.Sp）. Ossification was observed by saffrane-solid green staining， and serum levels of bone metabolism markers， including bone alkaline phosphatase （BALP）， osteocalcin （BGP）， type Ⅰ procollagen amino terminal propeptide （PINP）， and tartrate-resistant acid phosphatase 5b （TRACP-5b）， were determined by enzyme-linked immunosorbent assay （ELISA）. The protein and mRNA expression levels of Runx2， BMP-2， and Smad1 in rat femur were detected by Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. ResultCompared with the sham operation group， bone trabecula in the model group was sparse. BMD， BV/TV， Tb.N， and Tb.Th were decreased （P<0.05， P<0.01）. BS/BV （P<0.05） and Tb.Sp were increased. The content of BGP， BALP， PINP， and TRACP-5b in serum was significantly increased （P<0.01）. The mRNA and protein expressions of Runx2， BMP-2， and Smad1 in rat femur were significantly decreased （P<0.05， P<0.01）. Compared with the model group， the number of bone trabeculae in the high-dose and low-dose groups of Youguiwan was increased， and the bone microstructure was improved. BMD， BV/TV， Tb.N， and Tb.Th were increased significantly （P<0.05， P<0.01）， and BS/BV and Tb.Sp were increased. The content of bone metabolic markers decreased （P<0.05， P<0.01）. ConclusionYouguiwan has certain preventive and therapeutic effects on postmenopausal osteoporosis， and its mechanism may be related to promoting bone formation by regulating the BMP-2/Smad signaling pathway.

AIM:To explore the protective effect and mechanism of melittin against LPS induced acute lung injury.METHODS:Mice were intratrache-ally injected with LPS(5 mg/kg)to establish acute lung injury model.The lung coefficient of mice and the lung wet-dry mass ratio were determined.The lung tissue was stained with HE.The number of in-flammatory cells and expression levels of related in-flammatory factors in bronchoalveolar lavage fluid(BALF)were detected.The activities of serum SOD,MDA and GSH were detected.The expression of TLR4,NF-κB p65 and p-NF-κB p65 in lung tissues were determined by immunohistochemistry and Western blot.RESULTS:Melittin had a protective ef-fect on LPS-induced ALI,as evidenced by significant-ly reduced lung coefficient,lung wet/dry mass ra-tio,the number of inflammatory cells and the ex-pression level of inflammatory factors in BALF,and improved pulmonary pathology(P＜0.05,P＜0.01).Compared with model group,the serum MDA level of melittin group was significantly decreased(P＜0.01),while the activities of SOD and GSH were sig-nificantly increased(P＜0.05,P＜0.01).In addition,melittin can significantly reduce the expression of TLR4,inhibit the phosphorylation of NF-κB p65,and reduce the inflammatory response(P＜0.01).CONCLUSION:Melittin can significantly improve LPS-induced ALI,and the protective effect of melit-tin is mainly related to the inhibition of TLR4/NF-κB signaling pathway and the reduction of oxidative stress and inflammation.

AIM:To explore the protective effect and mechanism of melittin against LPS induced acute lung injury.METHODS:Mice were intratrache-ally injected with LPS(5 mg/kg)to establish acute lung injury model.The lung coefficient of mice and the lung wet-dry mass ratio were determined.The lung tissue was stained with HE.The number of in-flammatory cells and expression levels of related in-flammatory factors in bronchoalveolar lavage fluid(BALF)were detected.The activities of serum SOD,MDA and GSH were detected.The expression of TLR4,NF-κB p65 and p-NF-κB p65 in lung tissues were determined by immunohistochemistry and Western blot.RESULTS:Melittin had a protective ef-fect on LPS-induced ALI,as evidenced by significant-ly reduced lung coefficient,lung wet/dry mass ra-tio,the number of inflammatory cells and the ex-pression level of inflammatory factors in BALF,and improved pulmonary pathology(P＜0.05,P＜0.01).Compared with model group,the serum MDA level of melittin group was significantly decreased(P＜0.01),while the activities of SOD and GSH were sig-nificantly increased(P＜0.05,P＜0.01).In addition,melittin can significantly reduce the expression of TLR4,inhibit the phosphorylation of NF-κB p65,and reduce the inflammatory response(P＜0.01).CONCLUSION:Melittin can significantly improve LPS-induced ALI,and the protective effect of melit-tin is mainly related to the inhibition of TLR4/NF-κB signaling pathway and the reduction of oxidative stress and inflammation.

Objective:To evaluate the expressions of three biomarkers combination of CD27, CD38 and human leucocyte antigen (HLA)-DR in the application of discrminating active tuberculosis (ATB) and latent tuberculosis infection (LTBI).Methods:Sixty cases of ATB and 44 cases of LTBI were enrolled from March 2021 to February 2022 in Huashan Hospital, Fudan University and Wuxi Fifth People′s Hospital. Freshly isolated peripheral blood mononuclear cells (PBMC) from patients were stimulated with 6 kDa early secretory antigenic target/culture filtrate protein 10 peptide pools. The expressions of CD27, CD38 and HLA-DR on Mycobacterium tuberculosis-specific CD4 + T lymphocytes were evaluated by polychromatic flow cytometry. Mann-Whitney U test was used for statistical analysis. The area under the receiver operator characteristic curve (AUROC) was used to evaluate the diagnostic value of biomarkers in discriminating ATB and LTBI. Results:The frequencies of CD27 -, CD38 +, HLA-DR +, CD27 -CD38 +, CD27 -HLA-DR + and CD38 + HLA-DR + in ATB group were all higher than those in LTBI group, and the differences were all statistically significant ( U=26.00, 451.00, 384.00, 8.00, 7.00 and 184.00, respectively, all P<0.001). The AUROC of CD27 -CD4 + interferon-γ(IFN-γ) + T lymphocytes was 0.71 with a cut-off value of 52.31%, with the sensitivity of 50.00% and specificity of 87.20%. The AUROC of CD38 + CD4 + IFN-γ + T lymphocytes was 0.82 with a cut-off value of 30.25%, with the sensitivity of 73.40% and specificity of 89.70%. The AUROC of HLA-DR + CD4 + IFN-γ + T lymphocytes was 0.85 with a cut-off value of 36.60%, with the sensitivity of 66.00% and specificity of 94.90%. The AUROC of CD27 -CD38 + CD4 + IFN-γ + T lymphocytes was 0.80 with a cut-off value of 8.82%, with the sensitivity of 90.60% and specificity of 61.50%. The AUROC of CD27 -HLA-DR + CD4 + IFN-γ + T lymphocytes was 0.83 with a cut-off value of 18.62%, with the sensitivity of 75.00% and specificity of 79.50%. The AUROC of CD38 + HLA-DR + CD4 + IFN-γ + T lymphocytes was 0.93 with a cut-off value of 22.35%, with the sensitivity of 79.70% and specificity of 100.00%. Conclusions:The expressions of CD27 -, CD38 + and HLA-DR + in Mycobacterium tuberculosis-specific CD4 + T lymphocytes are higher in ATB group compared to LTBI group. ATB and LTBI could be well discriminated by detecting the expressions of CD27, CD38 and HLA-DR on CD4 + IFN-γ + T lymphocytes with flow cytometry.

With the continuous progress of research methodology in the real world and the growing maturity of artificial intelligence technology， a method for conducting “quantitative” research to guide clinical practice based on traditional Chinese medicine （TCM） diagnosis and treatment data was gradually developed. However， there is still a need for further improvements in the overall design of studies and the transformation of findings into clinical practice. Based on this， we put forward a comprehensive overall design concept and application approach for real-world study and artificial intelligence research based on clinical diagnosis and treatment data of TCM. This approach consists of five steps： Constructing a research-based database with a large sample size and high data quality； Mining and classification of core prescriptions； Conducting cohort studies to evaluate the effectiveness of core prescriptions； Utilizing case-control studies to clarify the dominant population； Establishing predictive models to achieve precision medicine. Additionally， it is imperative for researchers to establish a standardized system for collecting TCM variables and processing data， optimize the determination and measurement methods of confounding factors， further improve and promote methodologies， and strengthen the training of interdisciplinary talents. By following this research method， we anticipate that the clinical translation of research findings will be facilitated， leading to advancements in TCM precision medicine. Real-world study and artificial intelligence research share similar research foundations， and clinical applications complement each other. In the future， the two will merge together.

Objective:To investigate the role of glycoprotein A repetitions predominant (GARP) in the pathogenesis of tuberculosis through regulatory T cell (Treg), in order to provide new targets for the treatment of tuberculosis.Methods:Sixty patients with active pulmonary tuberculosis (ATB) admitted to Huashan Hospital, Fudan University and Wuxi Fifth People′s Hospital from January to September 2021 were included. And six individuals with latent tuberculosis infection (LTBI), and 16 healthy controls (HC) were recruited during the same period. Flow cytometry was performed to detect the proportion of Treg in the peripheral blood, and the expressions of GARP and transforming growth factor-β1 (TGF-β1) on Treg in different groups. Mann-Whitney U test was used for statistical analysis. Results:Among the 60 patients with ATB, 23 patients did not receive anti-tuberculosis drug therapy, 17 patients were treated for less than three months, ten patients were treated for three to less than six months, and ten patients were treated for greater than or equal to six months. The percentage of CD4 + CD25 + forkhead box protein 3 (Foxp3) + Treg in untreated ATB patients was 7.50%(5.67%, 9.00%), which was higher than that in HC (5.57%(5.03%, 6.09%)), and the difference was statistically significant ( U=95.00, P=0.010). The percentage of GARP expressing in CD4 + CD25 + Foxp3 + Treg in untreated ATB patients was 10.37%(7.79%, 12.90%), which was higher than that in LTBI (7.02%(5.15%, 8.81%)) and HC (5.33%(4.26%, 6.67%)), respectively, and the differences were both statistically significant ( U=31.00, P=0.040; U=36.00, P<0.001, respectively), while there was no significant difference between LTBI and HC ( U=25.00, P=0.095). The percentage of CD4 + CD25 + Foxp3 + Treg expressing TGF-β1 in untreated ATB patients was 7.13%(4.25%, 8.89%), which was higher than that in HC (3.59%(2.10%, 5.17%)), and the difference was statistically significant ( U=71.00, P=0.001). The expressions of GARP in CD4 + CD8 -CD25 + Foxp3 + Treg in patients with ATB treated for less than three months group, three to less than six months group and greater than or equal to six months group were 7.82%(3.94%, 13.17%), 6.92%(5.61%, 9.47%) and 7.26%(5.82%, 9.64%), respectively. The expressions of TGF-β1 in CD4 + CD8 -CD25 + Foxp3 + Treg in the above three treatment groups were 11.16%(7.91%, 15.23%), 8.66%(5.43%, 12.54%) and 7.82%(6.01%, 9.53%), respectively, and the expression of TGF-β1 in CD4 + CD8 -CD25 + Foxp3 + Treg in the patients with ATB treated for less than three months group was higher than that in the greater than or equal to six months group, the difference was statistically significant ( U=37.50, P=0.024). Conclusions:Foxp3/GARP/TGF-β1 pathway may be involved in the immune mechanism of Treg regulating the pathogenesis of tuberculosis, and GARP may be a new target for anti-tuberculosis therapy.

Objective We detected the level of serum calmodulin，γ-aminobutyric acid（GABA），glutamic acid（Glu） and GABA/Glu in insomnia patients，explored their association with insomnia.Methods The casecontrol study was used. This study comprised 119 insomnia patients and 122 healthy control subjects. Enzyme linked immunosorbent assay（ELISA） was utilized to measure the level of serum CaM，GABA，Glu and GABA/Glu. The correlation between the level of serum CaM and GABA/Glu was performed by correlation analyze. Results Our results suggested the level of serum CaM,GABA,GABA/Glu were significantly lower，and the level of serum Glu was significantly higher in patients with insomnia compared to the control group（P<0.05）.The level of serum CaM was positively correlated with GABA/Glu in insomnia patients（r=0.475，P<0.05）.Conclusion CaM downregulation increases the risk of insomnia，and the mechanism may be related to the downregulation of GABA/Glu level.

Objective:To investigate the diagnostic values of interleukin-22 (IL-22), interferon-γ(IFN-γ)and macrophage migration inhibition factor (MIF) in pleural effusion for tuberculosis pleurisy.Methods:From April 2018 to May 2019, a total of 77 patients including 45 cases of tuberculous pleurisy, 19 cases of malignant pleurisy, 13 cases of parapneumonia and 13 cases of healthy control in Wuxi Fifth People′s Hospital were enrolled. The levels of IL-22, IFN-γ and MIF in plasma and pleural effusion were detected by enzyme linked immunosorbent assay (ELISA). Mann-Whitney U test was used for statistical analysis.The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic values of IL-22, IFN-γ and MIF for tuberculous pleurisy. Results:The median levels of IL-22, IFN-γ, MIF and adenosine deaminase in 45 cases with pleural effusion in tuberculosis pleurisy group were 396.8 ng/L, 2 200.0 ng/L, 241.3 μg/L and 70.8 U/L, respectively, which were all significantly higher than 32 cases with non-tuberculosis pleurisy group, including 19 cases with malignant pleurisy and 13 cases with parapneumonia (52.8 ng/L, 232.3 ng/L, 179.6 μg/L and 17.0 U/L, respectively). The differences were all statistically significant ( U=179.000, 118.500, 287.000, 162.000, respectively, all P<0.05). The median levels of IL-22 and IFN-γ in plasma of tuberculosis pleurisy group were 20.0 ng/L and 45.9 ng/L, respectively, which were both higher than healthy control group (14.3 ng/L and 33.4 ng/L, respectively). The level of MIF was 96.2 μg/L, which was lower than healthy control (159.5 μg/L). The differences were all statistically significant ( U=74.000, 13.000 and 73.000, respectively, all P<0.05). The areas under ROC curve (AUC) of IL-22, IFN-γ and MIF in pleural effusion for the diagnosis of tuberculosis pleurisy were 0.876, 0.917 and 0.682, respectively.The sensitivities were 93.75%, 100.00% and 63.64%, respectively; the specificities were 82.22%, 91.11% and 65.85%, respectively. The median levels of IL-22 and IFN-γ in plasma in tuberculosis pleurisy group at two months of follow-up after anti-tuberculosis therapy were 16.0 ng/L and 33.9 ng/L, respectively, which were both lower than baseline (20.0 ng/L and 44.7 ng/L, respectively). The differences were both statistically significant ( U=2.156 and 2.221, respectively, both P<0.05). Conclusion:IFN-γ and IL-22 in pleural effusion could be used as effective indicators to identify tuberculous pleurisy, and the dynamic monitoring of IL-22 in patients′plasma could be an important biomarker in evaluating the efficacy of anti-tuberculosis treatment.

Objective To investigate the effect of dexmedetomidine hydrochloride ( Dex ) preconditioning on renal function in rats after renal ischemia reperfusion injury under high glucose condition. Methods SD rats were randomly divided into 6 groups:NG-Sham operated group,NG-I/R group, NG-Dex group,HG-Sham operated group,HG-I/R group,HG-Dex group. Renal ischemia reperfusion model was established except Sham groups. Dex 50 μg·kg-1 was injected intraperitoneally 30 min before ischemia in the Dex preconditioning group,25% glucose 3 g·kg-1 was given intraperitoneally before the renal ischemia reperfusion model was established in high glucose groups. Blood glucose and renal function of each group were detected . Renal pathologic changes were observed with hematoxylin-eosin staining. Apoptosis of renal tissue was detected by TUNEL method. Results BUN,Cr and apoptosis rate in NG-I/R group were higher than those in NG-Sham operated group ( P<0.05);BUN,Cr and apoptosis rate in NG-Dex group were lower than those in NG-I/R group ( P<0.05);BUN,Cr and apoptosis rate in HG-I/R group and HG-Dex group were higher than those in NG-I/R group and NG-Dex group,respectively (P<0.05); However,there was no significant difference between HG-I/R group and HG-Dex group ( P>0.05) . Conclusion Dex has a protective effect on renal function after renal ischemia reperfusion, but this effect is inhibited in high glucose condition, which may relate to the increasing of kidney cells apoptosis.

Objective:To investigate the effect of different ventilation time in the prone position on patients with endogenous/exogenous ARDS.Methods:30 endogenous/30 exogenous ARDS patients were randomly devided into 4 groups,ventilation in the prone position for 2 h and 4 h.Recording the score of APCHEII,oxygenation index,the absorption situation in X-ray,HR,MAP,extubation time,the time out of ICU.Results:The APCHEII scores HR and MAP in four groups have no significant statistics (P＞0.05);4h ventilation for endogenous ARDS patients has a better indicators than 2 h in oxygenation index,the absorption situation in X-ray,extubation time and the time out of ICU (P＜0.05);2 h and 4 h ventilation for exogenous ARDS patients can improve indicators above,two groups have no significant statistics (P＞0.05),the results of exogenous groups are precede than endogenous group (P＜0.05).Conclusion:Ventilation in the prone position can improve the situation of ARDS patients,both endogenous patients and exogenous patients.Exogenous ARDS patients have a better treatment effect after the ventilation of 2h,however,endogenous patients need longer time and have a non-ideal prognosis.