ObjectiveTo explore the possible mechanism of Tongbian Decoction （通便汤， TD） in treating slow transit constipation （STC）. MethodsTwenty-four SD rats were randomly divided into normal group， model group， TD group， and mosapride group， with 6 rats per group. Except for the normal group， STC models were established by intragastric administration of loperamide hydrochloride combined with normal saline. On the day following successful model establishment， rats in the TD group received 18.63 g·kg⁻¹ of TD by gavage， while those in the mosapride group received 1.605 mg·d⁻¹ of mosapride， and those in the normal group and the model group received 10 ml·kg⁻¹ of normal saline by gavage. All treatments were administered once daily for 7 consecutive days. Twenty-four hours after the last administration， fecal pellet number and fecal water content were measured. After intragastric administration of a 10% activated charcoal suspension， the small intestinal transit rate was calculated 30 minutes later. Serum levels of gastrin （GAS） and motilin （MTL） were measured by ELISA. Colonic histopathology was observed by HE staining， and mucus secretion by Alcian blue-periodic acid-Schiff （AB-PAS） staining. Ultrastructure of colon tissue was examined using transmission electron microscopy. Protein expression levels of C-kit， stem cell factor （SCF）， autophagy-related protein 5 （ATG5）， Beclin1， vesicle-associated membrane protein B （VAPB）， and protein tyrosine phosphatase interacting protein 51 （VAPB-PTPIP51） were measured by Western Blot， and the mRNA levels were detected by real-time PCR. Immunohistochemistry was used to detect SCF， C-kit， Beclin1， and ATG5 expression. The calcium content in colon tissue was determined by ELISA. ResultsCompared to the normal group， rats in the model group showed significantly reduced fecal pellet number， fecal water content， small intestinal transit rate， and serum GAS and MTL levels （P＜0.01）； the number of goblet cells decreased， and the mucosal and muscular layers of the colon became thinner； mRNA and protein expression levels of ATG5 and Beclin1 in colon tissue significantly increased， while calcium content decreased （P＜0.05 or P＜0.01）； and electron microscopy revealed vacuolar degeneration and increased autophagosomes in colonic cells. Compared to the model group， both TD group and mosapride group showed increased fecal pellet number， fecal water content， small intestinal transit rate， serum GAS and MTL levels， and colonic calcium content， along with decreased Beclin1 and ATG5 protein levels （P＜0.05 or P＜0.01）； the mucosal thickness and goblet cell number increased significantly， and autophagosomes decreased； in the TD group， ATG5 and Beclin1 mRNA levels decreased； in the mosapride group， SCF， VAPB， and PTPIP51 mRNA levels increased， while Beclin1 mRNA decreased （P＜0.05 or P＜0.01）. Compared to the mosapride group， the TD group showed higher fecal pellet number， fecal water content， serum GAS levels， colonic calcium content， and C-kit expression， along with lower ATG5 and Beclin1 levels （P＜0.05 or P＜0.01）. ConclusionTD may improve constipation symptoms by upregulating the VAPB-PTPIP51 complex during mitochondria-endoplasmic reticulum interactions， reducing autophagy of interstitial cells of Cajal， and promoting intestinal motility.

ObjectiveTo explore the possible mechanism of Tongbian Decoction （通便汤， TD） in treating slow transit constipation （STC）. MethodsTwenty-four SD rats were randomly divided into normal group， model group， TD group， and mosapride group， with 6 rats per group. Except for the normal group， STC models were established by intragastric administration of loperamide hydrochloride combined with normal saline. On the day following successful model establishment， rats in the TD group received 18.63 g·kg⁻¹ of TD by gavage， while those in the mosapride group received 1.605 mg·d⁻¹ of mosapride， and those in the normal group and the model group received 10 ml·kg⁻¹ of normal saline by gavage. All treatments were administered once daily for 7 consecutive days. Twenty-four hours after the last administration， fecal pellet number and fecal water content were measured. After intragastric administration of a 10% activated charcoal suspension， the small intestinal transit rate was calculated 30 minutes later. Serum levels of gastrin （GAS） and motilin （MTL） were measured by ELISA. Colonic histopathology was observed by HE staining， and mucus secretion by Alcian blue-periodic acid-Schiff （AB-PAS） staining. Ultrastructure of colon tissue was examined using transmission electron microscopy. Protein expression levels of C-kit， stem cell factor （SCF）， autophagy-related protein 5 （ATG5）， Beclin1， vesicle-associated membrane protein B （VAPB）， and protein tyrosine phosphatase interacting protein 51 （VAPB-PTPIP51） were measured by Western Blot， and the mRNA levels were detected by real-time PCR. Immunohistochemistry was used to detect SCF， C-kit， Beclin1， and ATG5 expression. The calcium content in colon tissue was determined by ELISA. ResultsCompared to the normal group， rats in the model group showed significantly reduced fecal pellet number， fecal water content， small intestinal transit rate， and serum GAS and MTL levels （P＜0.01）； the number of goblet cells decreased， and the mucosal and muscular layers of the colon became thinner； mRNA and protein expression levels of ATG5 and Beclin1 in colon tissue significantly increased， while calcium content decreased （P＜0.05 or P＜0.01）； and electron microscopy revealed vacuolar degeneration and increased autophagosomes in colonic cells. Compared to the model group， both TD group and mosapride group showed increased fecal pellet number， fecal water content， small intestinal transit rate， serum GAS and MTL levels， and colonic calcium content， along with decreased Beclin1 and ATG5 protein levels （P＜0.05 or P＜0.01）； the mucosal thickness and goblet cell number increased significantly， and autophagosomes decreased； in the TD group， ATG5 and Beclin1 mRNA levels decreased； in the mosapride group， SCF， VAPB， and PTPIP51 mRNA levels increased， while Beclin1 mRNA decreased （P＜0.05 or P＜0.01）. Compared to the mosapride group， the TD group showed higher fecal pellet number， fecal water content， serum GAS levels， colonic calcium content， and C-kit expression， along with lower ATG5 and Beclin1 levels （P＜0.05 or P＜0.01）. ConclusionTD may improve constipation symptoms by upregulating the VAPB-PTPIP51 complex during mitochondria-endoplasmic reticulum interactions， reducing autophagy of interstitial cells of Cajal， and promoting intestinal motility.

Objective: To accurately determine ABO blood groups through serological and molecular biological tests in patients whose forward typing was group O but who showed weak antibodies on one side of reverse typing, thereby ensuring precise transfusion. Methods: A total of 160 cases from 2023 to 2025 at Tianjin Blood Center, initially screened as O type with weak antibodies or missing antibodies on one side of reverse typing, were collected. Serological tests absorption-elution tests, and molecular biological tests were performed to accurately identify their ABO blood groups and ensure transfusion safety. Results: Among the 160 patients with forward typing as O type and weak antibodies on one side of reverse typing, 92 were identified as O type, 46 as ABO subtypes already named and recorded by ISBT (mainly including 18 cases of Ael02, 8 cases of AW37, and 9 cases of Bel03), 12 suspected as new alleles not yet recorded by ISBT (among which 5 were suspected to carry novel alleles that have not been recorded or reported in the literature), and 10 were suspected as weakened A/B antigens. Conclusion: In clinical transfusion, attention should be paid to cases with ABO forward-reverse discrepancy, where forward typing is O type and reverse typing shows weak antibodies on one side. These cases may involve ABO subtypes, and further testin is necessary to accurately determine the ABO blood group, achieve precise transfusion, and avoid blood wastage.

[Objective] To investigate the infection and characteristics of hepatitis E virus among blood donors in Hangzhou. [Methods] A total of 5 075 blood samples of blood donors from Zhejiang Provincial Blood Center from September to November 2023 were collected, including 5 037 samples with normal ALT and 38 samples with elevated ALT (>50 U/L). Enzyme-linked immunosorbent assay (ELISA) was used to detect anti-HEV IgM, anti-HEV IgG and HEV-Ag. The Fisher test and Chi-square test were used to evaluate the difference in the reactivity rates of anti-HEV IgM and anti-HEV IgG among different levels of ALT. The distribution characteristics of HEV screening in blood donors were analyzed. Univariate and multivariate logistic regression were used to analyze the susceptibility factors of anti-HEV IgM and anti-HEV IgG seropositivity, and the anti-HEV IgM-reactive blood donors were followed up by telephone. [Results] The reactivity rates of anti-HEV IgM, anti-HEV IgG and HEV-Ag in 5 075 blood samples were 0.45%, 22.98% and 0%, respectively. There was no difference in the reactivity rates of anti-HEV IgM and anti-HEV IgG among different levels of ALT (P>0.05), and the results of univariate and multivariate logistic regression analysis showed that age was a risk factor for anti-HEV IgM and anti-HEV IgG reactivity in blood donors (P<0.05), while no difference in the reactivity rates of anti-HEV IgM and anti-HEV IgG among blood donors was noticed in gender, occupation and education level (P>0.05). [Conclusion] There is a potential risk of transfusion-transmitted HEV (TT-HEV) in Hangzhou, and a cost-effective HEV screening strategy needs to be established to continue regular HEV surveillance in Hangzhou to assess the risk of infection.

Objective To compare and analyze the germicidal efficacy of continuous ultraviolet and pulsed ultravio-let(UV)lasers on pathogenic microorganisms.Methods Spore slides of Escherichia coli and Bacillus atrophaeus were irradiated using a 254 nm ultraviolet mercury lamp and UV laser.The carrier quantitative germicidal test was conducted to determine the disinfection efficacy at different irradiation doses.Results When the irradiation dose of the ultraviolet mercury lamp was 245.52 mJ/cm2,the logarithmic killing values of Escherichia coli and Bacillus atro-phaeus spores were 5.00 and 2.92,respectively,and the mean logarithmic killing doses were 49.10 and 84.08 mJ/cm2,respectively;When the UV laser irradiation doses were 208.39 and 206.80 mJ/cm2,the logarithmic killing values for the two microorganisms were 6.29 and 3.32,respectively,and the mean logarithmic killing doses were 33.13 and 62.29 mJ/cm2,respectively.Conclusion Compared with continuous UV radiation,pulsed UV laser has stron-ger penetration ability,better killing efficacy on pathogenic microorganisms at the same radiation dose,and can con-duct targeted disinfection and sterilization on the surface of objects directionally.

The immunosuppressive tumor micro-environment significantly limits the efficacy of im-munotherapy.Tumor-associated macrophages(TAMs),the most abundant immune cells in the tu-mor microenvironment,often exhibit an immuno-suppressive M2 phenotype,contributing to this im-munosuppressive landscape.Modulating TAMs to adopt anti-tumor phenotypes can enhance immu-notherapy outcomes and inhibit tumor progression.In recent years,tumor immunotherapy leveraging engineered bacteria has garnered considerable at-tention.Bacteria possess the ability to target tu-mors,preferentially colonizing tumor regions,and contain abundant pathogen-associated molecular patterns that effectively activate TAMs within the immunosuppressive tumor environment.This acti-vation enhances the tumoricidal and clearance ca-pabilities of TAMs.With the rapid advancements in synthetic biology,engineered bacteria have emerged as a potent therapeutic modality for im-munotherapy,leading to increased focus on the regulation of TAMs by engineered bacteria.This pa-per first outlines clinical studies on targeted TAMs therapy and engineered bacteria-based tumor ther-apy.It then reviews recent advancements in bacte-rial regulation of TAMs,detailing how engineered bacteria enhance TAM recruitment,improve TAM phagocytosis,and remodel TAM phenotypes.Mod-ulating TAMs with engineered bacteria presents a promising therapeutic strategy and introduces a novel approach in tumor immunotherapy.

The immunosuppressive tumor micro-environment significantly limits the efficacy of im-munotherapy.Tumor-associated macrophages(TAMs),the most abundant immune cells in the tu-mor microenvironment,often exhibit an immuno-suppressive M2 phenotype,contributing to this im-munosuppressive landscape.Modulating TAMs to adopt anti-tumor phenotypes can enhance immu-notherapy outcomes and inhibit tumor progression.In recent years,tumor immunotherapy leveraging engineered bacteria has garnered considerable at-tention.Bacteria possess the ability to target tu-mors,preferentially colonizing tumor regions,and contain abundant pathogen-associated molecular patterns that effectively activate TAMs within the immunosuppressive tumor environment.This acti-vation enhances the tumoricidal and clearance ca-pabilities of TAMs.With the rapid advancements in synthetic biology,engineered bacteria have emerged as a potent therapeutic modality for im-munotherapy,leading to increased focus on the regulation of TAMs by engineered bacteria.This pa-per first outlines clinical studies on targeted TAMs therapy and engineered bacteria-based tumor ther-apy.It then reviews recent advancements in bacte-rial regulation of TAMs,detailing how engineered bacteria enhance TAM recruitment,improve TAM phagocytosis,and remodel TAM phenotypes.Mod-ulating TAMs with engineered bacteria presents a promising therapeutic strategy and introduces a novel approach in tumor immunotherapy.

Objective:To explore the clinical efficacy of retrograde tibial intramedullary nail (RTN) combined with less invasive stabilization system (LISS) in the treatment of distal femoral fractures.Methods:A retrospective analysis was conducted on the data of 11 patients with distal femoral fractures who underwent RTN combined with LISS locking plate treatment at the 909th Hospital from June 2021 to December 2022. There were 5 males and 6 females, with an average age of 56.5±17.5 years. The fracture types were AO/OTA 33A2 in 2 cases, A3 in 2 cases, C1 in 2 cases, and C2 in 2 cases. There were 3 cases of periprosthetic femoral fractures after total knee arthroplasty (TKA), all classified as Rorabeck type Ⅱ. They were treated with retrograde intramedullary nailing of the tibia combined with a minimally invasive lateral internal fixation system. The operation time, intraoperative blood loss, fracture healing time, full weight-bearing time, femorotibial angle, range of motion (ROM) of the knee joint, Hospital for Special Surgery (HSS) score, visual analogue scale (VAS) for pain, and complications were recorded.Results:The operation was successfully completed in all 11 cases. The operation time was 98.3±9.0 min (range 83-115 min), and the intraoperative blood loss was 167.8±24.3 ml (range 120-210 ml). All 11 cases were followed up for 11.0±1.9 months (range 9-15 months). The healing time of 11 cases was 5.3±0.8 months (range 4-6 months) after operation. The time of complete weight-bearing activity after operation was 55.7±6.5 d (range 46-67 d). At the last follow-up, the femoral-tibial angle of 11 cases was 171.2°±1.8° (range 169°-174°), the ROM of knee joint was 129.5°±4.7° (range 120°-135°), and the HSS score was 86.8±6.9 points (range 69-95 points). There were no major complications except for one case of superficial infection of surgical incision after operation.Conclusion:RTN combined with lateral LISS locking plate showed good clinical effect with the advantages of less trauma, reliable fixation, rapid recovery, less postoperative complications.

Objective To study how lipid bilayer fluidity modulates the interaction between β1 integrin and CD40L,as well as the formation of CD40L-mediated tumor cell contact interfaces.Methods Supported lipid bilayers(SLB)with different fluidities were prepared through adjusting the 1,2-dioleoyl-sn-glycero-3-[N-(5-amino-l-carboxypentyl)iminodiacetic acid]succinyl nickel salt(DGS-NTA)content.The functionalization of lipid bilayers was achieved by anchoring fluorescently labeled CD40L molecules onto the membrane surface.The contact interface formation of PC9 cells on the functionalized lipid bilayers was observed through confocal fluorescence imaging and fluorescence recovery after photobleaching(FRAP)experiments,and data of two dimensional(2D)reaction kinetics of β1 integrin and CD40L were extracted from Zhu-Golan plots.Results The diffusion coefficient of molecules in lipid bilayer was negatively correlated with DGS-NTA content.High fluidity of lipid bilayer promoted CD40L accumulation at cell contact interface and expanded the cell contact area.The 2D dissociation constants(2D Kd)of β1 integrin-CD40L complexes were approximately 13,31 and 65 molecules/μm2 for the three lipid bilayers with high,moderate and low fluidities,respectively.Conclusions High fluidity of lipid bilayers significantly facilitates diffusion and aggregation of CD40L to the cell contact interface,thus enhancing β1 integrin-CD40L interaction and the stability of cell contact interfaces.

Objective:To investigate the clinicopathological and genetic features of infantile rhabdomyofibrosarcoma (IRFS) with EGFR kinase domain duplication (EGFR-KDD).Methods:The clinical, morphological and immunohistochemical features of three IRFS with EGFR-KDD diagnosed from January 2022 to January 2024 at Department of Pathology, Foshan Traditional Chinese Medicine Hospital, Foshan, China were retrospectively analyzed using PCR or next generation sequencing technique; and related literature was reviewed.Results:There were 1 male and 2 females, aged at presentation ranging from 1 to 4 years. The tumor occurred in the left thigh, right maxillofacial region, and right popliteal space. The presenting symptom was a painless mass which was accidentally discovered. The maximum diameter of tumors ranged from 3 to 5 cm. Microscopically, the tumors were poorly defined and composed of relatively monomorphic spindle cells, arranged in diffuse, fascicular growth patterns, with moderate pale eosinophilic cytoplasm. Mitoses were abundant. A few round rhabdomyoblastic tumor cells with abundant eosinophilic cytoplasm were found. There was no evidence of hemorrhage or necrosis. The tumor cells expressed vimentin, SMA, MSA, desmin, MyoD1 and myogenin; and the Ki-67 proliferation index was 10%-60%. RT-PCR showed EGFR-KDD in all three cases. Gene fusion was detected in three cases based on next generation sequencing, but only one case had EGFR-KDD. Follow-up data for 12 to 36 months showed two patients died of the disease and one patient was alive without recurrences and metastasis.Conclusions:IRFS is a rare soft tissue tumor that resembles infantile fibrosarcoma but has immunohistochemical evidence of rhabdomyoblastic differentiation. It more commonly occurs in infants and tends to appear in limbs and torso with poor prognosis. Aggressive multimodality treatment is recommended for these patients. EGFR-KDD may be a genetic driver to IRFS. Clinical response to EGFR targeted therapy might be promising in the future.