Objective To investigate whether the extracts of Patrinia heterophylla(MTH)inhibits the activation of macrophages induced by lipopolysaccharide(LPS)in vitro and the inflammatory response induced in vivo by suppressing the nuclear factor kappa B(NF-κB)signaling pathway.Methods Cellular studies:RAW264.7 cells were divided into 6 groups:control group,LPS group,LPS+Dexamethasone(DEX)group and LPS+MTH(low,medium,high dose)groups.The LPS group,LPS+DEX and LPS+MTH group were induced with a final concentration of 100 ng/ml LPS.While the LPS+DEX group was additionally treated with 5.0 ng/ml DEX,the LPS+MTH group was additionally treated with MTH(final concentrations of 0.1,0.2 and 0.4 mg/ml).The cell activity was detected using CCK-8 assay,cell invasion was detected using Transwell assay,cell apoptosis was detected using flow cytometry,and interleukin-6(IL-6),interleukin-17(IL-17)and tumor necrosis factor-α(TNF-α)were detected using real-time fluorescence quantitative PCR and enzyme-linked immunosorbent assay,respectively.The expression and secretion levels of nitric oxide(NO)in cells were detected by Griess assay,total reactive oxygen species(ROS)levels were detected by flow cytometry.The phosphorylation level of p65 and inhibitor of κB(IκB)were detected by protein immunoblotting assay.The transcriptional activity was detection by luciferase reporter gene assay.Animal studies:50 rats were randomly divided into 5 groups,10 per group,namely the control group,LPS group,LPS+DEX group,LPS+MTH low-dose group(6 g/kg)and LPS+MTH high-dose group(24 g/kg).LPS was injected into the lungs of rats,and the groups were orally administered at 36 h,24 h,12 h before modeling,and 12 h,24 h after modeling.12 h after the last administration,bronchoalveolar lavage fluid was collected,and the IL-6,IL-17,TNF-α and NF-κB signaling pathway-related indicators in the bronchoalveolar lavage fluid were measured.Results Cellular studies:Compared with the control group,the cell activity,invasion,apoptosis,IL-6,IL-17 and TNF-α mRNA and protein,NO,ROS,phosphorylation level of P536 protein S536 site and IκB protein S32 site and transcription activity of NF-κB had significantly increased in the LPS group(all P＜0.05).Compared with the LPS group,the cell activity,invasion,IL-6,IL-17,and TNF-αmRNA and protein,NO,ROS,phosphorylation level of P536 protein S536 site and IκB protein S32 site and transcription activity of NF-κB had significantly decreased in 3 LPS+MTH groups(all P＜0.05),and apoptosis was significantly increased in 3 LPS+MTH groups(all P＜0.05).All of which showed a dose-dependent trend of MTH.Animal studies:Compared with the control group,the LPS group showed a significant increase in IL-6,IL-17,TNF-α and phosphorylation levels of p65 protein at S536 site and IκB protein at S32 site(all P＜0.05).Compared with the LPS group,the 2 LPS+MTH groups showed a significant decrease in IL-6,IL-17,TNF-α and phosphorylation levels of p65 protein at S536 site and IκB protein at S32 site(all P＜0.05).These indicators showed a dose-dependent trend with MTH.Conclusion MTH can inhibit the activation of mouse macrophage RAW264.7 induced by LPS and the inflammatory response in the lungs of rats induced by LPS,which may be related to the inhibition of the NF-κB signaling pathway.

Objective:This study aims to summarize the theoretical basis and practical effect of interdisciplinary team building in the National Clinical Medical Research Center for Aging and Medicine, Huashan Hospital.Methods:Interactive memory was employed as the theoretical basis for building cross-disciplinary teams, and the work was carried out in three dimensions of specialization, credibility, and coordination.Results:The center had shaped a coordinative team atmosphere and strengthened its integrity in five dimensions, including discipline construction, open platform, innovation curation, science popularization, and technology transfer. The center also promoted the implementation of research tasks in the three core areas.Conclusions:As the aging healthcare issues are highly diversified, complex, and systematic, the center needs to continuously improve its ability to build interdisciplinary teams and provide a platform for integrating various types of resources into the solution of healthcare problems of the elderly.

Objective To investigate the influencing factors of peripheral infections of knee joint tumor prosthesis as well as the value of serum D-D and TLR2 in predicting the infection risks so as to provide a reference for early diagnosis of tumorous periprosthetic infection(PJI)of knee joint.Methods The patients who were treated and followed up in our department from January 2008 to June 2020 were selected.According to the inclusion and exclusion criteria,136 of the patients were selected.The data including age,gender,BMI,history of diabetes,smoking history,tumor location,stage of malignant tumor,operation time,osteotomy length,intraoperative bleed-ing,and the percentage of neutrophils,leukocytes,serum D-dimer,and serum TLR value 3 days after operation were collected.The risk factors of PJI and the diagnostic value of serum D-dimer and serum TLR were analyzed.Results The incidence of PJI was 11.76%.Postoperative chemotherapy and operation time≥180 min were the risk factors of PJI(P<0.05).The area under curve(AUC)of the combination of two indicators,serum D-dimer and serum TLR2 were 0.917,0.894 and 0.778,respectively.The AUC of TLR2 was lower than that of the combina-tion of two indicators(P<0.05);The sensitivity was 0.975,0.908 and 0.708,respectively,and the specificity was 0.75,0.75,and 0.812,respectively.Conclusion Postoperative chemotherapy and operation time≥180 min are the risk factors of PJI.The combination of D-dimer and TLR2 has good diagnostic value.

OBJECTIVE To explore the effect of acupuncture on the learning and memory ability and intestinal flora structure of senescence-accelerated mouse prone 8(SAMP8)mice.METHODS Eighteen 6-month-old SAMP8 mice were randomly divided in-to model group and acupuncture group,with 9 mice in each group,and 9 senescence-accelerated mouse resistant strain 1(SAMR1)mice of the same month as the normal group.The acupuncture group selected Baihui and Yongquan points for acupuncture,5 times a week,20 min each time,for 8 consecutive weeks.Morris water maze was used to detect the learning and memory ability of mice in each group,immunofluorescence method was used to detect β-amyloid protein(Aβ)expression in the cerebral cortex,and 16S rDNA technology was used to detect changes in intestinal flora.RESULTS The results of the water maze experiment showed that compared with the normal group,the learning and memory abilities of the mice in the model group were significantly reduced(P<0.05),and the learning and memory abilities of the mice in the acupuncture group were significantly improved compared with the model group(P<0.05).Immunofluorescence results showed that compared with the normal group,the model group had increased positive expression of Aβ in the cerebral cortex,and obvious Aβ plaque deposition could be seen;the acupuncture group had fewer Aβ plaques than the model group.Alpha diversity index showed that compared with the normal group,the Ace and Simpson values of the model group de-creased(P<0.05)and the Shannon value increased(P<0.05);compared with the model group,the Ace and Chao values of the acu-puncture group increased(P<0.05,P<0.01).In the beta diversity analysis,the results of principal component analysis(PCA)and principal coordinate analysis(PCoA)showed that the samples in each group were relatively clustered,and the distance between sam-ples in each group was far away.The Venn diagram showed that there were 664,689,and 649 OTUs in the normal group,model group,and acupuncture group,respectively.Results of intestinal flora structure analysis showed that compared with the normal group,the abundance of Bacteroidetes,Ruminococcaceae,and Eubacterium xylanophilum in the model group increased(P<0.05),and the abundance of Fusobacterium,unclassified Fusobacterium,and Porphyromonas decreased(P<0.05);compared with the model group,the abundance of Bacteroidetes and Eubacterium xylanophilum decreased in the acupuncture group(P<0.05),and the abundance of Fusobacterium,unclassified Fusobacterium,and Porphyromonas increased(P<0.05).CONCLUSION Acupuncture can reduce cortical Aβ deposition and improve learning and memory abilities in SAMP8 mice,which may be related to regulating the structure of intestinal flora and increasing species diversity.

Objective To investigate whether the extracts of Patrinia heterophylla(MTH)inhibits the activation of macrophages induced by lipopolysaccharide(LPS)in vitro and the inflammatory response induced in vivo by suppressing the nuclear factor kappa B(NF-κB)signaling pathway.Methods Cellular studies:RAW264.7 cells were divided into 6 groups:control group,LPS group,LPS+Dexamethasone(DEX)group and LPS+MTH(low,medium,high dose)groups.The LPS group,LPS+DEX and LPS+MTH group were induced with a final concentration of 100 ng/ml LPS.While the LPS+DEX group was additionally treated with 5.0 ng/ml DEX,the LPS+MTH group was additionally treated with MTH(final concentrations of 0.1,0.2 and 0.4 mg/ml).The cell activity was detected using CCK-8 assay,cell invasion was detected using Transwell assay,cell apoptosis was detected using flow cytometry,and interleukin-6(IL-6),interleukin-17(IL-17)and tumor necrosis factor-α(TNF-α)were detected using real-time fluorescence quantitative PCR and enzyme-linked immunosorbent assay,respectively.The expression and secretion levels of nitric oxide(NO)in cells were detected by Griess assay,total reactive oxygen species(ROS)levels were detected by flow cytometry.The phosphorylation level of p65 and inhibitor of κB(IκB)were detected by protein immunoblotting assay.The transcriptional activity was detection by luciferase reporter gene assay.Animal studies:50 rats were randomly divided into 5 groups,10 per group,namely the control group,LPS group,LPS+DEX group,LPS+MTH low-dose group(6 g/kg)and LPS+MTH high-dose group(24 g/kg).LPS was injected into the lungs of rats,and the groups were orally administered at 36 h,24 h,12 h before modeling,and 12 h,24 h after modeling.12 h after the last administration,bronchoalveolar lavage fluid was collected,and the IL-6,IL-17,TNF-α and NF-κB signaling pathway-related indicators in the bronchoalveolar lavage fluid were measured.Results Cellular studies:Compared with the control group,the cell activity,invasion,apoptosis,IL-6,IL-17 and TNF-α mRNA and protein,NO,ROS,phosphorylation level of P536 protein S536 site and IκB protein S32 site and transcription activity of NF-κB had significantly increased in the LPS group(all P＜0.05).Compared with the LPS group,the cell activity,invasion,IL-6,IL-17,and TNF-αmRNA and protein,NO,ROS,phosphorylation level of P536 protein S536 site and IκB protein S32 site and transcription activity of NF-κB had significantly decreased in 3 LPS+MTH groups(all P＜0.05),and apoptosis was significantly increased in 3 LPS+MTH groups(all P＜0.05).All of which showed a dose-dependent trend of MTH.Animal studies:Compared with the control group,the LPS group showed a significant increase in IL-6,IL-17,TNF-α and phosphorylation levels of p65 protein at S536 site and IκB protein at S32 site(all P＜0.05).Compared with the LPS group,the 2 LPS+MTH groups showed a significant decrease in IL-6,IL-17,TNF-α and phosphorylation levels of p65 protein at S536 site and IκB protein at S32 site(all P＜0.05).These indicators showed a dose-dependent trend with MTH.Conclusion MTH can inhibit the activation of mouse macrophage RAW264.7 induced by LPS and the inflammatory response in the lungs of rats induced by LPS,which may be related to the inhibition of the NF-κB signaling pathway.

Objectives: . We aimed to develop a new calculation model for calcium requirements in dialysis patients following parathyroidectomy. Methods: . A total of 98 patients with secondary hyperparathyroidism receiving parathyroidectomy from January 2014 to January 2022 were enrolled in this study. Among these patients, 78 were randomly selected for construction of the calcium requirement calculation model, and the remaining 20 patients were selected for model validation. The calcium requirement model estimated the total calcium supplementation for 1 week after surgery using variables with significant relationships in the derivation group by stepwise multiple linear regression analysis. Bias, precision, and accuracy were measured in the validation group to determine the performance of the model. Results: . The model was as follows: calcium requirement for 1 week after surgery=33.798–8.929×immediate postoperative calcium+0.190×C-reactive protein–0.125×age+0.002×preoperative intact parathyroid hormone+0.003×preoperative alkaline phosphatase (R2=0.8). The model was successfully validated. Conclusion . We generated a novel model to guide calcium supplementation. This model can assist in stabilizing the serum calcium levels of patients during the early postoperative period. Furthermore, it contributes to the individualized and precise treatment of hypocalcemia in patients following parathyroidectomy.

Objective:To explore the effects of dietary phosphate restriction education on serum phosphorus level, dietary phosphate intake and the knowledge of hyperphosphatemia in maintenance hemodialysis (MHD) patients.Methods:This study was a retrospective cohort study. A total of 116 hemodialysis patients in Huashan Hospital, Huadong Hospital and Tongji Hospital from October 2019 to December 2020 were enrolled in this study. They were divided into short-term group (84 cases) and long-term group (32 cases). The short-term group did not receive education or received education≤60 minutes. Meanwhile, the long-term group received education>60 minutes. Serum phosphorus level, dietary phosphate intake and knowledge of hyperphosphatemia were compared between the two groups after 4 weeks.Results:At baseline, age [64(56, 69) years old vs 65(60, 73) years old, Z=-1.493, P=0.136], the proportion of males [58.3%(49/84) vs 56.3%(18/32), χ2=0.041, P=0.839], dialysis age [55(26, 130) months vs 53(20, 132) months, Z=-0.062, P=0.951], body mass index, diabetes history, single-pool Kt/V, proportion of calctriol used, blood calcium, blood phosphorus, intact parathyroid hormone and dietary protein, dietary phosphorus and dietary phosphorus protein ratio had no statistical significance between short-term group and long-term group (all P>0.05). Adequate dietary phosphate restriction education reduced dietary phosphate intake [777.98(653.81, 943.16) mg/d vs 896.56(801.51, 1 015.51) mg/d, Z=-2.903, P=0.004], phosphate/protein ratio [13.16(11.52, 14.21) mg/g vs 15.27(13.31, 17.48) mg/g, Z=-3.929, P<0.001] and serum phosphorus level [(1.42±0.37) mmol/L vs (1.85±0.44) mmol/L, t=4.984, P<0.001]. Meanwhile, such education significantly improved achievement rate of serum phosphorus (62.5% vs 41.7%, χ2=4.034, P=0.045). In addition, patients in long-term group answered more questions correctly (completely correct plus partially correct) about the causes (93.8% vs 72.6%, χ2=6.120, P=0.013), poor prognosis (78.1% vs 52.4%, χ2=6.372, P=0.012) of hyperphosphatemia as well as the types of food with high phosphate (65.6% vs 52.4%, χ2=1.650, P=0.199). Conclusion:Adequate dietary phosphate restriction education reduces serum phosphorus level and dietary phosphate intake, and improves the knowledge of hyperphosphatemia in MHD patients.

Objective:To screen and identify H-2 d-restricted T cell epitopes in fusion (F) and attachment (G) glycoproteins of Nipah virus (NiV) in mice. Methods:The complete peptides (single peptide contains 15 amino acids, and 10 amino acids were repeated in the front and back peptides) derived from F and G antigens were mixed into peptide libraries. BALB/c mice were immunized with DNA vaccines expressing NiV F and G proteins alone and in combination. The full sequence peptide libraries of F and G antigens were mixed into peptide pools by matrix design, and spleen cells of immunized mice were collected and analyzed by IFN-γ ELISPOT assay to detect the dominant H-2 d-restricted epitope peptides. Results:Twelve dominant H-2 d-restricted peptides were screened from the F protein-specific peptide library and the 56th peptide produced the strongest reaction. Four dominant peptides were screened from the G protein-specific peptide library and the 72nd peptide produced the strongest reaction. Conclusions:In this study, 12 F antigen-specific and 4 G antigen-specific H-2 d restricted dominant T cell epitopes of NiV were screened and identified by IFN-γ ELISPOT, which could provide reference for immunological analysis of NiV and vaccine research.

Objective:To establish an in vitro culture system of small intestinal organoid in normal mice and perform functional identification, and to provide an in vitro research tool for material transport in the intestine under chronic kidney disease. Methods:The small intestinal crypts of C57BL/6J mice were isolated, extracted and cultured in an in vitro three dimension culture system. The formation of small intestinal organoid was observed with inverted microscope. The tissue structure of the small intestinal organoid was observed by hematoxylin and eosin staining. The cellular composition of the small intestinal organoid was identified by immunofluorescence. The expression of substance absorption-related transporters in the small intestinal organoid was detected by real time fluorescence quantitative PCR. Results:The small intestinal crypts were successfully extracted. The organoids of small intestine and different intestinal segments were successfully constructed. The cultured organoids had vigorous proliferation ability and maintained proliferation ability after passing through generations. Immunofluorescence results showed that the small intestinal organoids expressed mucin2, chromogranin A, oflm4 and lysozyme, which were different types of intestinal cell biomarkers. The PCR results showed that small intestinal organoids expressed calcium, phosphate and sodium absorption-related transporters, and the mRNA expression levels of major transporters for sodium and phosphate absorption in different intestinal segments-like organs were consistent with those in vivo, which was consistent with the characteristics of small intestinal segmental absorption. Conclusions:The successful construction of small intestine and different intestinal segments organoids, and the first observation of the expression of substance absorption-related transporters in such organoid, provide a stable and convenient in vitro research tool for the development of intestinal substance transport in chronic kidney disease.

Closed-loop hospital management can effectivly cope with the COVID-19 pandemic. In order to ensure the continuity of treatments for hemodialysis patients under closed-loop management and minimize possible medical and infection risks, Huashan Hospital affiliated to Fudan University and 9 hospitals in Shanghai established a hemodialysis alliance in January 2021.The alliance optimized hemodialysis resources within the region through overall planning by preparing sites, materials and personnel shifts in advance, and establishing management systems and work processes to ensure that patients could be quickly and orderly diverted to other blood dialysis centers for uninterrupted high-quality hemodialysis services, in case that some hemodialysis centers in the alliance under closed-loop management.From November 2021 to April 2022, 317 of 1 459 hemodialysis patients in the alliance were diverted to other centers for treatment, accumulating 1 215 times/cases of treatments without obvious adverse reactions. The practice could provide a reference for medical institutions to quickly establish mutual support mode under major public health events.