ObjectiveTo improve the therapeutic effect of Buyang Huanwutang（BYHW） on diabetic kidney disease （DKD） and explore new methods for developing new Chinese medicine decoctions，we utilized 5-hydroxymethylcytosine （5hmC）-Seal sequencing technology and network pharmacology to modify BYHW. MethodsWe selected 14 diabetes mellitus （DM） patients and 15 DKD patients hospitalized in the Department of Endocrinology of Peking University Third Hospital in 2021. Circulating free DNA （cfDNA） in the patients’ plasma was sequenced. After data processing and screening， we performed temporal clustering analysis to select a DKD 5hmC gene set， which was then cross-validated with a DKD database gene set to obtain the DKD gene set. We retrieved target genes of the seven herbal components of BYHW from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and the Encyclopedia of Traditional Chinese Medicine （ETCM）， and performed cross-analysis with the DKD gene set to identify common genes shared by the disease and the Chinese medicines. A protein-protein interaction （PPI） network was constructed for the common genes to screen out the key genes. Chinese medicines targeting these key genes were searched against ETCM to identify removable Chinese medicines. Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis was performed on non-common DKD genes， and key genes in DKD-related pathways were selected based on machine learning. The GSE30529 dataset was used to verify the expression trends of 5hmC-modified genes and the feasibility of target genes as drug targets. TCMBank was used to search for target genes and obtain compounds targeting these genes and the corresponding Chinese medicines to construct a "key target-compound-Chinese medicine" network. Molecular docking was employed to verify the binding affinity of compounds with key targets. TCMSP and ETCM were used to search and count the candidate Chinese medicines targeting DKD-related genes， and a new decoction was formed by adding the selected Chinese medicines. A mouse model of DKD was established to examine the efficacy of the new decoction based on the mouse body mass， random blood glucose， urinary microalbumin （mALB）， serum creatinine （Scr）， and blood urea nitrogen （BUN） and by hematoxylin-eosin staining， periodic acid-Schiff staining， Masson staining， immunofluorescence assay， and Real-time PCR. ResultsThe cross-analysis results showed that the DKD gene set included 507 genes， of which 30 were target genes of BYHW. The PPI analysis indicated that the top 15% target genes regarding the degree were interleukin-6 （IL-6）， Toll-like receptor 4 （TLR4）， lactotransferrin （LTF）， lipoprotein lipase （LPL）， and sterol regulatory element-binding transcription factor 1 （SREBF1）. Persicae Semen and Pheretima in BYHW were unrelated to key genes and removed. Machine learning identified 10 potential target genes， among which TBC1 domain family member 5 （TBC1D5）， RAD51 paralog B （RAD51B）， and proteasome 20S subunit alpha 6 （PSMA6） had expression trends consistent with the GSE30529 dataset and could serve as drug targets. The "key target-compound-Chinese medicine" network and molecular docking results indicated that the compounds with good binding affinity to target proteins were arginine， glycine， myristicin， serine， and tyrosine， corresponding to 121 Chinese medicines. The top 10 Chinese medicines targeting DKD-related genes were Lycii Fructus， Ginseng Radix et Rhizoma， Dioscoreae Rhizoma， Rehmanniae Radix Praeparata， Isatidis Radix， Glehniae Radix， Ophiopogonis Radix， Allii Sativi Bulbus， Isatidis Folium， and Bolbostemmatis Rhizoma. Based on traditional Chinese medicine theory， the new decoction was obtained after removal of Persicae Semen and Pheretima and addition of Rehmanniae Radix Praeparata and Dioscoreae Rhizoma. Animal experiment results indicated that the modified BYHW improved the kidney function and inhibited renal fibrosis in DKD mice， with better effects than the original decoction. ConclusionThe BYHW modified based on 5hmC-Seal sequencing demonstrates better performance in inhibiting fibrosis and ameliorating DKD than the original decoction. This elucidates the biomedical theory behind the epigenetic modification of traditional Chinese medicine prescriptions， potentially offering new perspectives for the exploration of these prescriptions

ObjectiveTo improve the therapeutic effect of Buyang Huanwutang（BYHW） on diabetic kidney disease （DKD） and explore new methods for developing new Chinese medicine decoctions，we utilized 5-hydroxymethylcytosine （5hmC）-Seal sequencing technology and network pharmacology to modify BYHW. MethodsWe selected 14 diabetes mellitus （DM） patients and 15 DKD patients hospitalized in the Department of Endocrinology of Peking University Third Hospital in 2021. Circulating free DNA （cfDNA） in the patients’ plasma was sequenced. After data processing and screening， we performed temporal clustering analysis to select a DKD 5hmC gene set， which was then cross-validated with a DKD database gene set to obtain the DKD gene set. We retrieved target genes of the seven herbal components of BYHW from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and the Encyclopedia of Traditional Chinese Medicine （ETCM）， and performed cross-analysis with the DKD gene set to identify common genes shared by the disease and the Chinese medicines. A protein-protein interaction （PPI） network was constructed for the common genes to screen out the key genes. Chinese medicines targeting these key genes were searched against ETCM to identify removable Chinese medicines. Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis was performed on non-common DKD genes， and key genes in DKD-related pathways were selected based on machine learning. The GSE30529 dataset was used to verify the expression trends of 5hmC-modified genes and the feasibility of target genes as drug targets. TCMBank was used to search for target genes and obtain compounds targeting these genes and the corresponding Chinese medicines to construct a "key target-compound-Chinese medicine" network. Molecular docking was employed to verify the binding affinity of compounds with key targets. TCMSP and ETCM were used to search and count the candidate Chinese medicines targeting DKD-related genes， and a new decoction was formed by adding the selected Chinese medicines. A mouse model of DKD was established to examine the efficacy of the new decoction based on the mouse body mass， random blood glucose， urinary microalbumin （mALB）， serum creatinine （Scr）， and blood urea nitrogen （BUN） and by hematoxylin-eosin staining， periodic acid-Schiff staining， Masson staining， immunofluorescence assay， and Real-time PCR. ResultsThe cross-analysis results showed that the DKD gene set included 507 genes， of which 30 were target genes of BYHW. The PPI analysis indicated that the top 15% target genes regarding the degree were interleukin-6 （IL-6）， Toll-like receptor 4 （TLR4）， lactotransferrin （LTF）， lipoprotein lipase （LPL）， and sterol regulatory element-binding transcription factor 1 （SREBF1）. Persicae Semen and Pheretima in BYHW were unrelated to key genes and removed. Machine learning identified 10 potential target genes， among which TBC1 domain family member 5 （TBC1D5）， RAD51 paralog B （RAD51B）， and proteasome 20S subunit alpha 6 （PSMA6） had expression trends consistent with the GSE30529 dataset and could serve as drug targets. The "key target-compound-Chinese medicine" network and molecular docking results indicated that the compounds with good binding affinity to target proteins were arginine， glycine， myristicin， serine， and tyrosine， corresponding to 121 Chinese medicines. The top 10 Chinese medicines targeting DKD-related genes were Lycii Fructus， Ginseng Radix et Rhizoma， Dioscoreae Rhizoma， Rehmanniae Radix Praeparata， Isatidis Radix， Glehniae Radix， Ophiopogonis Radix， Allii Sativi Bulbus， Isatidis Folium， and Bolbostemmatis Rhizoma. Based on traditional Chinese medicine theory， the new decoction was obtained after removal of Persicae Semen and Pheretima and addition of Rehmanniae Radix Praeparata and Dioscoreae Rhizoma. Animal experiment results indicated that the modified BYHW improved the kidney function and inhibited renal fibrosis in DKD mice， with better effects than the original decoction. ConclusionThe BYHW modified based on 5hmC-Seal sequencing demonstrates better performance in inhibiting fibrosis and ameliorating DKD than the original decoction. This elucidates the biomedical theory behind the epigenetic modification of traditional Chinese medicine prescriptions， potentially offering new perspectives for the exploration of these prescriptions

Periodontitis is a common oral disease caused by bacteria coupled with an excessive host immune response. Stem cell therapy can be a promising treatment strategy for periodontitis, but the relevant mechanism is complicated. This study aimed to explore the therapeutic potential of mitochondria from human embryonic stem cell-derived mesenchymal stem cells (hESC-MSCs) for the treatment of periodontitis. The gingival tissues of periodontitis patients are characterized by abnormal mitochondrial structure. Human gingival fibroblasts (HGFs) were exposed to 5 μg/mL lipopolysaccharide (LPS) for 24 h to establish a cell injury model. When treated with hESC-MSCs or mitochondria derived from hESC-MSCs, HGFs showed reduced expression of inflammatory genes, increased adenosine triphosphate (ATP) level, decreased reactive oxygen species (ROS) production, and enhanced mitochondrial function compared to the control. The average efficiency of isolated mitochondrial transfer by hESC-MSCs was determined to be 8.93%. Besides, a therapy of local mitochondrial injection in mice with LPS-induced periodontitis showed a reduction in inflammatory gene expression, as well as an increase in both the mitochondrial number and the aspect ratio in gingival tissues. In conclusion, our results indicate that mitochondria derived from hESC-MSCs can reduce the inflammatory response and improve mitochondrial function in HGFs, suggesting that the transfer of mitochondria between hESC-MSCs and HGFs serves as a potential mechanism underlying the therapeutic effect of stem cells.
Humans ; Gingiva/cytology* ; Fibroblasts/metabolism* ; Mitochondria/physiology* ; Mesenchymal Stem Cells/cytology* ; Animals ; Periodontitis/therapy* ; Mice ; Reactive Oxygen Species/metabolism* ; Inflammation ; Lipopolysaccharides ; Human Embryonic Stem Cells/cytology* ; Cells, Cultured ; Adenosine Triphosphate/metabolism* ; Male

Objective To explore the regulatory effect of long non-coding RNA HOX transcript anti-sense RNA(lnRNA HOTAIR)targeting tuberous sclerosis complex 1(TSC1)on podocyte injury in diabetic nephropathy(DN).Methods Mice podocyte MPC5 were divided into 6 groups:the control group,the high glucose group(the HG group),the HG+si-NC group,the HG+si-HOTAIR group,the HG+si-HOTAIR+sh-NC group,and the HG+si-HOTAIR+sh-TSC1 group.qPCR was used to detect the levels of lnRNA HO-TAIR and TSC1 mRNA.Cell viability was detected by cell counting kit-8(CCK-8),and apoptosis was detec-ted by flow cytometry.ELISA was used to detect the levels of inflammatory factors[IL-6,tumor necrosis fac-tor-α(TNF-α)]and the levels of oxidative stress indicators[reactive oxygen species(ROS),malondialdehyde(MDA),superoxide dismutase(SOD)]were detected by corresponding reagent kit.Results Compared with the control group,the levels of lnRNA HOTAIR,apoptosis rate,IL-6,TNF-α,ROS and MDA in the HG group were significantly increased(P<0.05),while the levels of TSC1 mRNA,cell viability and SOD were significantly decreased(P<0.05).Compared with the HG+si-NC group,the levels of lnRNA HOTAIR,ap-optosis rate,IL-6,TNF-α,ROS and MDA in the HG+si-HOTAIR group significantly decreased(P<0.05),the levels of TSC1 mRNA,cell viability and SOD were significant increased(P<0.05).Compared with the HG+si-HOTAIR+sh-NC group,the levels of apoptosis rate,IL-6,TNF-α,ROS and MDA in the HG+si-HOTAIR+sh-TSC1 group were significantly increased(P<0.05),the levels of TSC1 mRNA,cell viability and SOD were significantly decreased(P<0.05).Conclusion Silencing lnRNA HOTAIR can target TSC1 to regulate the activity,apoptosis rate,inflammatory level and oxidative stress levels of podocyte injury in DN,thereby alleviating podocyte damage in DN.

T cell receptor (TCR) is a key molecule mediating anti-tumor immunity, and its diversity profile (TCR repertoire) serves as a biomarker of host immune status. The development of high-throughput sequencing technologies has revolutionized the application of TCR repertoire analysis in cancer immunotherapy. The clinical value of TCR sequencing in individualized tumor treatment is increasingly prominent. Through monitoring immunotherapy response and predicting survival outcomes, TCR sequencing provides critical guidance for precision tumor therapy.

Objective To develop a modified lentiviral expression system of mCherry-GFP-LC3B,driven by the hematopoietic-specific SFFV(spleen focus-forming virus)promoter,in order to perform an efficient and real-time monitoring of autophagy flux with in terminally differentiated erythroid cells.Methods The lentiviral plasmid pRSC-SFFV-mCherry-GFP-LC3B was constructed and packaged into lentiviruses for infection of human erythroid progenitor cell line(HUDEP-2)and mouse fetal liver-derived primary erythroid cells.Autophagy dynamics of the cells were then analyzed using fluorescence imaging,Western blot,and flow cytometry in serum starvation and chloroquine inter-vention models.Results The SFFV promoter rendered significantly higher reporter expression efficiency than CMV promoter in HUDEP-2(97%vs.60%)(P＜0.01)and in the primary mouse erythroid cells(83%vs.1%)(P＜0.001),without disrupting normal erythroid differentiation.Serum deprivation increased au-tolysosomes(red puncta),elevated LC3-Ⅱ/LC3-Ⅰratios,and decreased p62 levels.Chloroquine treatment in-duced autophagosome(yellow puncta)accumulation(P＜0.001),showing a dose-dependent inhibition of auto-phagy flux(r2=0.92).Conclusions The SFFV-driven dual-fluorescent system enables robust and real-time mo-nitoring of autophagy flux in erythroid cells,providing a sensitive tool for mechanistic study of erythroid differenti-ation and related disorders such as anemia.

Objective　To timely understand the current status and distribution of nail snails (Oncomelania hupensis) in Tongjiang River channels in Zhenjiang City, providing a scientific basis for achieving the goal of eliminating schistosomiasis. Methods　From 2019 to 2023, nine Tongjiang River channels on the south bank of the Yangtze River under the jurisdiction of Zhenjiang City were selected as the monitoring area. Snail monitoring was carried out onshore and beach nail snails floats in the Tongjiang River channels, nail snails on attachments in hardened areas, snails induced by straw curtains, and snails carried by boats and domestic animals. Results　The monitoring results of shoreline snail from 2019 to 2023 showed that the snail situation in the Tongjiang River channel and its outer river bank remained relatively stable from 2019 to 2020; however, in 2021, under the influence of the Yangtze River flooding disaster in 2020, the area of snails increased significantly. In 2021, the area with snails in the Tongjiang River channel and the outer river bank increased by 45.70% (11.95/26.15) and 100.00% (20.00/20.00) compared to 2020; the average density of nails snails in the Tongjiang River channel and the outer river bank and the emergence rate of snail frames both showed a significant increase, rising by 94.73% (0.18/0.19) and 68.08% (8.68/12.75) compared to 2020, and by 122.73% (0.81/0.66) and 102.78% (43.26/42.09), respectively. The differences in the increase in the occurrence rate of spiked frames in the Tongjiang River channel Chili River and Renmin River were not statistically significant (χ2=0.329, P>0.05; χ2=0.646, P>0.05). From 2022 to 2023, the density of nail snails and the occurrence rate of framed snails in the Tongjiang River channel showed a decreasing trend (F=4.72, P=0.04 and χ2=372.58, P<0.01). The area of nail snails, density of live snails, and occurrence rate of framed snails in the outer river bank showed a decreasing trend (F=13.96, P=0.02; F=23.43, P<0.01; χ2=1 029.69, P<0.01). During the five years, no nail snails were detected in the ancient canal and 11 tributaries. From 2019 to 2023, 180 times of 3 003 kg of floating objects were salvaged, with a total of 148 live snails detected. A total of 17 live snails were captured on attachments in the hardened berm area; a total of 11 live snails were captured by straw curtain snail baiting;112 boats were inspected, and no snails were found; there were 112 boats surveyed, and no snails were found; 97 cattle were observed, and 2 cattle were found to carry 1 live snail on their hooves; 321 sheep were observed, and no snails were found on their hooves; and no infectious snails were found in the monitoring area in 5 years. Conclusions　Nail snails continue to exist in the Tongjiang River channel, and the risk factors for schistosomiasis transmission have not been completely eliminated. It is still necessary to carry out accurate monitoring of the snail situation in the Tongjiang River and the river bank, so as to grasp the risk of transmission in time and take emergency measures.

Objective:To analyze the short-term clinical outcomes of total laparoscopic distal gastrectomy (TLDG) and laparoscopic-assisted distal gastrectomy (LADG) combined with Billroth-Ⅱ+Braun anastomosis.Methods:Clinical characteristics of patients undergoing laparoscopic distal gastrectomy combined with Billroth-Ⅱ+Braun anastomosis at Department of Gastrointestinal Surgery, Shandong Provincial Hospital Affiliated to Shandong First Medical University from Jan 2020 to Oct 2022 were analyzed. Patients were divided into TLDG group ( n=62) and LADG group ( n=62) according to the surgical approach. Results:There were significant differences in the preoperative clinical data section between the two groups, and 124 patients (62 in each group) were enrolled after using propensity score matching to balance significant variables. Compared with the LADG group, the TLDG group showed statistically differences in time to first venting [(2.9±1.3) vs. (2.3±0.8) d, Z=-3.072, P=0.002], time to first fluid diet [(5.9±1.3) vs. (5.4±1.4) d, Z=-2.031, P=0.042] and incision length [(7.1±1.4) vs. (4.8±0.8) cm, Z=-6.331, P=0.000]. Total postoperative complication rate in the TLDG group and the LADG group (29% vs. 37%, χ2=0.911, P=0.340) was not statistically significant. Incidence of postoperative pneumonia was lower in the TLDG group than in the LADG group (3% vs. 13%, χ2=3.916, P=0.048), and incidence of all remaining postoperative complications were not statistically significant. There was no statistically significant difference in the incidence of serious postoperative complications between the TLDG and LADG groups ( P=1.000). Multifactorial analysis revealed that male ( P=0.023) and age ≥65 years ( P=0.001) were independent risk factors for postoperative complications. Conclusion:TLDG is safe and feasible and has better short-term clinical efficacy than LADG.

Objective To investigate the changes in distribution of Oncomelania hupensis snails in forestlands in Songjiang District, Shanghai Municipality from 2009 to 2023, so as to provide insights into formulation of O. hupensis snail surveillance programs. Methods The reports on O. hupensis snail surveillance in Songjiang District, Shanghai Municipality from 2009 to 2023 were collected, and the snail surveillance data in forestlands were extracted. The trends in the proportion of areas with snails in forestlands in total areas with snails, occurrence of frames with living snails and density of living snails were evaluated using a Joinpoint regression model in Songjiang District from 2009 to 2023, and the annual percent change (APC) and average annual percent change (AAPC). Results A total of 40 sites with snails were found in forestlands in 14 administrative villages of 4 townships, Songjiang District, Shanghai Municipality from 2009 to 2023. A total of 39 065 frames were surveyed for snails in settings covering an area of 609 600 m², and there were 6 084 frames with snails, covering 151 250 m² snail habitats. A total of 22 210 snails were captured, with the highest density of 260.00 snails/0.1 m², and 6 262 snails were dissected, with no Schistosoma japonicum infection identified in snails. The proportion of areas with snails in forestlands in total areas with snails appeared a tendency towards a rise in forestlands in Songjiang District, Shanghai Municipality from 2009 to 2023 (APC = AAPC = 24.9%, P > 0.05); however, there were no turning points in the trend curve, with the highest proportion seen in 2009 (53.81%), the lowest in 2011 and 2023 (both 0) and a mean proportion of 24.81%. The occurrence of frames with living snails appeared a tendency towards a rise from 2009 to 2023 (APC = AAPC = 41.5%, P > 0.05); however, there were no turning points in the trend curve, with the highest occurrence in 2009 (53.81%), the lowest in 2011 and 2013 (both 0), and the mean occurrence of 15.57%. In addition, the density of living snails appeared a tendency towards a rise from 2009 to 2023 (APC = AAPC = 55.0%, P > 0.05); however, there were no turning points in the trend curve, with the highest density in 2023 (0.96 snails/0.1 m²), the lowest in 2011 and 2013 (both 0), and a mean density of 0.57 snails/0.1 m². Conclusions The difficulty in O. hupensis snail control and risk of imported snails appeared a tendency towards a rise in forestlands in Songjiang District, Shanghai Municipality over years from 2009 to 2023. Supervision and assessment prior to seedling transplantation and intensified surveillance post-transplantation are recommended to reduce the risk of O. hupensis snail importation and spread.

ObjectiveTo analyze the monitoring results and trends of Oncomelania in Songjiang District of Shanghai from 2007 to 2023， and to provide reference for future monitoring work. MethodsThe data of snail monitoring in Songjiang District from 2007 to 2023 were collected， including the location of snail spots， the number of survey frames， the area with snails， the number of frames with snails， the total number of snails， etc. The density of living snails and the occurrence rate of living snail frames were calculated. ResultsFrom 2007 to 2023， a total of 158 snail spots were found in 22 administrative villages in 4 towns in Songjiang District， including 131 emerging snail habitats （82.91%） and 27 reemerging snail habitats （17.09%）. The total snail area was 175 980 m²， the number of snail frames was 10 061， and 33 302 snails were captured. The highest density was 260/0.11 m²， and 15 662 snails were dissected. No positive snails were found. The average density of living snails was 0.60/0.11 m² and the average occurrence rate of living snail frames was 17.99%. The snail areas in different areas were Yexie Town 126 000 m² （71.60%）， Maogang Town 26 470 m² （15.04%）， Xinbang Town 17 040 m² （9.68%）， and Shihudang Town 6 470 m² （3.68%）. The average density of living snails in different areas was 0.89/0.11 m² in Yexie Town， 0.65/0.11 m² in Maogang Town， 0.26/0.11 m² in Shihudang Town and 0.24/0.11 m² in Xinbang Town. The snail areas of different snail habitats were as follows： woodland 151 250 m²（85.95%）， river 13 810 m²（7.85%）， ditch 6 910 m²（3.93%）， farmland 4 010 m²（2.28%）. The average density of living snails in different snail habitats： ditch 1.01/0.11 m²， beach 0.86/0.11 m²， woodland 0.57/0.11 m²， river 0.40/0.11 m². The occurrence rate of living snail frames in different regions and different snail environment types showed a downward trend and the difference was statistically significant. ConclusionAlthough the snail monitoring indicators such as snail area， average living snail density and average living snail frame occurrence rate in Songjiang District are still at its historically low level， the average living snail density and average living snail frame occurrence rate are still high compared with other regions or the national average. We should focus on the upper reaches and tributaries of the Huangpu River， especially the key areas of seedling transplantation， and strengthen monitoring and supervision to reduce the risk of snail input.