Objective:To investigate the predictive value of systemic immune inflammation index for the efficacyof neoadjuvant chemotherapy in triple negative breast cancer patients, and analyzed the relationship between pathological complete response (pCR) and prognosis.Methods:The clinical data of 146 patients with triple-negative breast cancer admitted to the 926th Hospital of the Joint Service Support Force of the PLA from January 2018 to December 2020 were retrospectively collected. All patients received neoadjuvant chemotherapy. After chemotherapy, the patients were divided into pCR group (62 cases) and non-pCR group (84 cases) according to whether the patients achieved pCR. Pathological characteristics and systemic immunoinflammatory index levels of the two groups were compared. Receiver operating characteristic (ROC) curve was used to analyze the predictive value of systemic immunoinflammatory index for pCR after neoadjuvant chemotherapy in patients with triple-negative breast cancer, and survival curves were drawn to compare the disease-free survival of the two groups.Results:The rate of axillary lymph node metastasis in pCR group was lower than that in non-pCR group: 37.10% (23/62) vs. 64.29% (54/84), there was statistical difference ( χ2 = 10.58, P<0.01). There were no significant differences in TNM stage, Ki-67 level and histological grade between the two groups ( P>0.05). Compared with the non -pCR group, the systemic immune inflammation index in the pCR group was significantly reduced: 617.42 ± 166.40 vs. 853.67 ± 202.41, P<0.01. Systemic immune inflammation index was valuable in predicting non-pCR of triple negative breast cancer patients after neoadjuvant chemotherapy, and the area under the curve was 0.807 (95% CI: 0.738 - 0.875, P<0.01). Compared with the non-pCR group, the disease-free survival of patients in the pCR group was significantly prolonged ( P = 0.033). Conclusions:Systemic immune inflammation index was related to the efficacy of neoadjuvant chemotherapy in triple negative breast cancer patients, and can be used as a biological indicator to predict the efficacy of neoadjuvant chemotherapy in triple negative breast cancer.

IGF-1 is involved in the growth and development of mammals,but its role in sebaceous gland excretion,B16 cell proliferation,and migration in the skin has not been reported yet.This study aims to reveal the function of IGF-1 by detecting its expression in animal skin.Using nano-body screening and purification methods,IGF-1 nanobodies(IGF-1-VHH)were obtained.Using IGF-1-VHH as the primary antibody,Western blot and immunohistochemistry were used to detect the expression of IGF-1 in bovine skin and alpaca acne.IGF-1-VHH was added to melanoma B16 cell culture medium,and CCK-8 and scratch healing methods were used to detect the effects of IGF-1 nanobodies on B16 cell proliferation and migration,as well as their possible molecular mech-anisms.The results showed that the obtained IGF-1-VHH could be applied in immunohistochemis-try and Western blot detection methods,with strong IGF-1 positive expression signals in bovine sebaceous glands and alpaca acne.IGF-1-VHH binds to IGF-1 in cells and has a certain inhibitory effect on the proliferation and migration of B16 cells by regulating the expression of RAS,ERK,and RAF.In summary,IGF-1 maybe involved in the secretion and excretion of sebum in animal skin.IGF-1-VHH inhibits the proliferation and migration of B16 cells by binding to IGF-1,provi-ding a new theoretical basis for ensuring normal physiological function of the skin and clinical di-agnosis and treatment of melanoma.

Objective:To establish and optimize a novel method, focus forming assay (FFA), for quantitation of influenza A virus (FluA) and compare its application performance with traditional plague forming assay (PFA).Methods:The foci chromogenic effects of three peroxidase substrates in immunostaining were compared. The PFA and FFA methods were used to explore FluA incubation times and plaque morphology on 12-well plates, and to determine optimal incubation times and virus adsorption volumes for different FluA subtypes on 96-well plates. The correlation between FFA and PFA was evaluated, and the optimized FFA was applied to the in vitro antiviral efficacy analysis of Favipiravir and neutralization test against different subtypes of FluA. Results:TRUEBLUE substrate was identified as the optimal substrate for foci visualization. Compared with the PFA, the FFA showed improved sensitivity and reduced detection time in FluA titration, and good correlation was shown between the two methods′ results. By replacing the 96-well plate with the 12-well plate for FFA titration of different subtypes of FluA, the detection time was shortened, and the amount of serum samples used could be further reduced by optimizing the virus adsorption volume. The half-maximal effective concentration of favipiravir against influenza viruses assessed by the FFA and PFA methods showed no significant difference, and was consistent with the results obtained from quantitative PCR. Additionally, the focus reduction neutralization test and hemagglutination inhibition assays demonstrated strong correlation in determining antibody titers against FluA in serum neutralization assays.Conclusions:The improved FFA method developed here provides a more efficient experimental tool for FluA titration, antiviral drug screening and broad-spectrum vaccine evaluation.

Objective:To establish and optimize a novel method, focus forming assay (FFA), for quantitation of influenza A virus (FluA) and compare its application performance with traditional plague forming assay (PFA).Methods:The foci chromogenic effects of three peroxidase substrates in immunostaining were compared. The PFA and FFA methods were used to explore FluA incubation times and plaque morphology on 12-well plates, and to determine optimal incubation times and virus adsorption volumes for different FluA subtypes on 96-well plates. The correlation between FFA and PFA was evaluated, and the optimized FFA was applied to the in vitro antiviral efficacy analysis of Favipiravir and neutralization test against different subtypes of FluA. Results:TRUEBLUE substrate was identified as the optimal substrate for foci visualization. Compared with the PFA, the FFA showed improved sensitivity and reduced detection time in FluA titration, and good correlation was shown between the two methods′ results. By replacing the 96-well plate with the 12-well plate for FFA titration of different subtypes of FluA, the detection time was shortened, and the amount of serum samples used could be further reduced by optimizing the virus adsorption volume. The half-maximal effective concentration of favipiravir against influenza viruses assessed by the FFA and PFA methods showed no significant difference, and was consistent with the results obtained from quantitative PCR. Additionally, the focus reduction neutralization test and hemagglutination inhibition assays demonstrated strong correlation in determining antibody titers against FluA in serum neutralization assays.Conclusions:The improved FFA method developed here provides a more efficient experimental tool for FluA titration, antiviral drug screening and broad-spectrum vaccine evaluation.

Human alphaherpesvirus 2 (HSV-2) is the main pathogen resulting human genital herpes, which poses a major threat to the socio-economic development, while there is no effective vaccine. In this study, we developed a novel lipopolyplex (LPP)-delivered mRNA vaccine expressing the HSV-2 envelope glycoprotein gD and evaluated its immunogenicity in mice. The mRNA vaccine was prepared from the genetically modified gD mRNA synthesized in vitro combined with the LPP delivery platform and it was named gD-ORI mRNA. The expression of gD antigen in the mRNA vaccine was validated in vitro by Western blotting and indirect immunofluorescence assay, then the immune responses induced by this mRNA vaccine in mice were evaluated. The immunization with gD mRNA alone induced strong humoral and cellular immune responses in mice. Robust and long-lasting gD-specific IgG antibodies were detected in the mouse serum after booster immunization with gD-ORI mRNA. The immunized mice exhibited a Th1/Th2 balanced IgG response and robust neutralizing antibodies against HSV-2, and a clear dose-response relationship was observed. The gD-specific IgG antibodies were maintained in mice for a long time, up to 18 weeks post-booster immunization. At the same time, multifunctional gD-specific CD4⁺ and CD8⁺ T cells in vaccinated mice were detected by intracellular cytokine staining (ICS). This novel gD-expressing mRNA vaccine delivered by LPP induces strong and long-lasting immune responses in mice post booster immunization and has a promising prospect for development and application. This study provides scientific evidence and reference for the development of a new mRNA vaccine for HSV-2.
Animals ; Herpesvirus 2, Human/genetics* ; Viral Envelope Proteins/genetics* ; Mice ; Herpes Genitalis/immunology* ; RNA, Messenger/immunology* ; Female ; Mice, Inbred BALB C ; Antibodies, Viral/blood* ; mRNA Vaccines/immunology* ; Antibodies, Neutralizing/blood* ; Humans

Objective:To explore the application value of three-dimensional high-resolution anorectal manometry(3D-HRAM)model in the diagnosis and localization of anal sphincter injury after surgery for complex anal fistula.Methods:A prospective study was con-ducted on 92 patients who had undergone surgery for complex anal fistula in the Department of Coloproctology of The People's Hospi-tal Affiliated to Fujian University of Traditional Chinese Medicine from January 2023 to January 2024.The study was conducted in two stages.In the first stage,92 patients underwent 3D-HRAM and endoanal ultrasonography(EAUS)to examine the sphincter,and the data were compared with intraoperative records to evaluate the diagnostic efficiency and localization accuracy of 3D-HRAM and EAUS.In the second stage,92 patients who had been examined were grouped according to the presence or absence of symptoms of anal dysfunction to further verify the diagnostic and localization values of 3D-HRAM and EAUS for postoperative compensated/decompen-sated sphincter injury.Results:Among 92 patients,41 and 57 patients were diagnosed with sphincter injury through 3D-HRAM and EAUS,with a sensitivity of 61.19%and 85.07%,a specificity of 88%and 76%,an area under the curve of 0.739 and 0.813 calcu-lated by plotting the receiver operating characteristic curve,and a localization accuracy of 44.77%and 71.64%,respectively,showing significant differences between 3D-HRAM and EAUS(P=0.002),indicating that 3D-HRAM had lower diagnostic efficiency and local-ization accuracy than EAUS for anal sphincter injury after surgery for complex anal fistula.After grouping by symptoms,the diagnostic efficiency and localization accuracy of 3D-HRAM for symptomatic sphincter injury(decompensated)were significantly better than those for asymptomatic sphincter injury,and were similar to those of EAUS(P=0.763).Conclusion:The application advantage of 3D-HRAM lies in the diagnosis and localization of decompensated anal sphincter injury after surgery for complex anal fistula,which provides effective technical support for the study of anal defecation control mechanisms and injury compensation mechanisms.

IGF-1 is involved in the growth and development of mammals,but its role in sebaceous gland excretion,B16 cell proliferation,and migration in the skin has not been reported yet.This study aims to reveal the function of IGF-1 by detecting its expression in animal skin.Using nano-body screening and purification methods,IGF-1 nanobodies(IGF-1-VHH)were obtained.Using IGF-1-VHH as the primary antibody,Western blot and immunohistochemistry were used to detect the expression of IGF-1 in bovine skin and alpaca acne.IGF-1-VHH was added to melanoma B16 cell culture medium,and CCK-8 and scratch healing methods were used to detect the effects of IGF-1 nanobodies on B16 cell proliferation and migration,as well as their possible molecular mech-anisms.The results showed that the obtained IGF-1-VHH could be applied in immunohistochemis-try and Western blot detection methods,with strong IGF-1 positive expression signals in bovine sebaceous glands and alpaca acne.IGF-1-VHH binds to IGF-1 in cells and has a certain inhibitory effect on the proliferation and migration of B16 cells by regulating the expression of RAS,ERK,and RAF.In summary,IGF-1 maybe involved in the secretion and excretion of sebum in animal skin.IGF-1-VHH inhibits the proliferation and migration of B16 cells by binding to IGF-1,provi-ding a new theoretical basis for ensuring normal physiological function of the skin and clinical di-agnosis and treatment of melanoma.

Objective:To investigate the predictive value of systemic immune inflammation index for the efficacyof neoadjuvant chemotherapy in triple negative breast cancer patients, and analyzed the relationship between pathological complete response (pCR) and prognosis.Methods:The clinical data of 146 patients with triple-negative breast cancer admitted to the 926th Hospital of the Joint Service Support Force of the PLA from January 2018 to December 2020 were retrospectively collected. All patients received neoadjuvant chemotherapy. After chemotherapy, the patients were divided into pCR group (62 cases) and non-pCR group (84 cases) according to whether the patients achieved pCR. Pathological characteristics and systemic immunoinflammatory index levels of the two groups were compared. Receiver operating characteristic (ROC) curve was used to analyze the predictive value of systemic immunoinflammatory index for pCR after neoadjuvant chemotherapy in patients with triple-negative breast cancer, and survival curves were drawn to compare the disease-free survival of the two groups.Results:The rate of axillary lymph node metastasis in pCR group was lower than that in non-pCR group: 37.10% (23/62) vs. 64.29% (54/84), there was statistical difference ( χ2 = 10.58, P<0.01). There were no significant differences in TNM stage, Ki-67 level and histological grade between the two groups ( P>0.05). Compared with the non -pCR group, the systemic immune inflammation index in the pCR group was significantly reduced: 617.42 ± 166.40 vs. 853.67 ± 202.41, P<0.01. Systemic immune inflammation index was valuable in predicting non-pCR of triple negative breast cancer patients after neoadjuvant chemotherapy, and the area under the curve was 0.807 (95% CI: 0.738 - 0.875, P<0.01). Compared with the non-pCR group, the disease-free survival of patients in the pCR group was significantly prolonged ( P = 0.033). Conclusions:Systemic immune inflammation index was related to the efficacy of neoadjuvant chemotherapy in triple negative breast cancer patients, and can be used as a biological indicator to predict the efficacy of neoadjuvant chemotherapy in triple negative breast cancer.

Objective:Clinical characteristics and diagnosis and treatment process was reported and analyzed of a patient with brucellosis complicated with thyroid abscess, providing reference for the clinical diagnosis of brucellosis complicated with thyroid abscess.Methods:Clinical medical records of a patient with brucellosis complicated with thyroid abscess who was treated at the General Surgery Department of Yanchi County People's Hospital in Wuzhong City, Ningxia Hui Autonomous Region in November 2021 were collected. The clinical manifestations, blood routine, brucella antibodies, thyroid function, bacterial culture, thyroid ultrasound and other examination results, as well as the diagnosis and treatment process, were comprehensively analyzed. Results:The patient was a male, 61 years old, who presented with a neck mass without typical clinical manifestations of brucellosis. Thyroid ultrasound revealed a space occupying lesion, and the preliminary diagnosis was thyroid cystadenoma. Thyroid right lobe and isthmus resection surgery was performed. During the operation, it was found that some of the thyroid glands were tightly adhered to the cervical blood vessels, so the resection surgery was changed to abscess drainage, and the drainage fluid was purulent and bloody. The bacterial culture result of thyroid purulent fluid (intraoperative puncture fluid and postoperative drainage fluid) was brucella lamblia, and the serum brucella test tube agglutination test titer was 1 ∶ 400 (+++). The patient improved and was discharged after local drainage and anti brucella treatment. Follow up for 4 months showed no abnormalities. Conclusions:Brucellosis which begins with a local infection of the thyroid gland is extremely rare, with no characteristic clinical manifestations, and is prone to misdiagnosis. Timely correction of the surgical plan during the treatment process avoids the removal of the patient's thyroid, which has a certain clinical reference value.

Objective:To construct a novel respiratory syncytial virus (RSV) vaccine based on a recombinant influenza virus vector and evaluate its immune protective effects in mice.Methods:A recombinant H1N1 influenza A virus (IAV) expressing the extracellular domain (Gecto) of RSV A2 G protein was constructed and rescued, named as PR8NAGecto/WSN. After in vitro verification of the Gecto expression and PR8NAGecto/WSN growth kinetics, a single dose of PR8NAGecto/WSN was used to immunize BALB/c mice through intranasal administration to evaluate the efficacy of PR8NAGecto/WSN by assessing humoral (IgG, neutralizing antibody), mucosal (IgA) and cellular immunity (IFN-γ ELISPOT). Four weeks after immunization, the mice were challenged with RSV A2 or RSV B9320 to evaluate the protective effects of PR8NAGecto/WSN by analyzing mouse body weight changes, lung tissue virus titers and pathological changes. Results:A single-dose intranasal immunization with PR8NAGecto/WSN induced robust humoral, mucosal and cellular immunity in mice. Moreover, the mice in the immunized group had lower lung virus loads and mild lung pathological damages following the challenge with RSV A or RSV B subtype as compared with the control group.Conclusions:A single-dose intranasal immunization with PR8NAGecto/WSN induces robust immunity and provide protection against RSV A and B challenges in mice. This study provides new ideas and reference for the development of novel mucosal vaccines against RSV.