ObjectiveTo investigate the protective effects and mechanisms of Bushen Zhuyun prescription （BSZY） on abortion rats with kidney deficiency-corpus luteum inhibition syndrome. MethodsAn abortion rat model with kidney deficiency-corpus luteum inhibition syndrome was constructed. Pregnant mice aged 8-10 weeks were randomly divided into a control group （Control）， a model group （Model）， low-dose BSZY （BSZY-L）， medium-dose BSZY （BSZY-M）， and high-dose BSZY （BSZY-H） groups （2.57， 5.14， 10.28 g·kg^-¹）， and a Zishen Yutai Pill （ZSYT） group （1.575 g·kg^-¹）. Hematoxylin-eosin （HE） staining was used to evaluate histopathological changes in ovarian and decidual tissue of rats in each group. Enzyme-linked immunosorbent assay （ELISA） was employed to measure levels of estrogen （E₂）， progesterone （P）， luteinizing hormone （LH）， prolactin （PRL）， and follicle-stimulating hormone （FSH） in serum. The candidate targets of BSZY were obtained from the Traditional Chinese Medicine System Pharmacology Platform （TCMSP） and Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine （TCMIP） v2.0 databases， while disease targets for recurrent spontaneous abortion （RSA） were retrieved from GeneCards， DrugBank， Online Mendelian Inheritance in Man （OMIM）， and Therapeutic Target Database （TTD）. The intersection targets were identified by the Venny 2.1.0 platform. Pathway enrichment analysis was conducted based on the Metascape database to predict the potential mechanisms of BSZY. Additionally. Western blot was used to verify the effects of BSZY on the expression of estrogen receptor （ERα）， phosphatidylinositol 3-kinase （PI3K）， and protein kinase B （Akt） and explore its protective mechanism on RSA rats. ResultsCompared with the control group， the model group exhibited significantly decreased uterine， ovarian， and embryonic wet weights （P<0.05， P<0.01）， with an abortion rate of 57.18%. The ovarian tissue showed varying degrees of reduction in primordial follicles， primary follicles， mature follicles， and corpora lutea， along with a large number of atretic follicles. The endometrium was thinner， and decidual tissue exhibited cellular edema and disorganized arrangement. In contrast， compared with the model group， the BSZY groups at all doses and the ZSYT group demonstrated increased uterine， ovarian， and embryonic wet weights， along with a reduced abortion rate. The number of primordial follicles， primary follicles， mature follicles， and corpora lutea increased， while atretic follicles decreased. The endometrium thickened， and decidual tissue displayed normal cellular structure with tight arrangement. Additionally， the model group showed significantly decreased levels of E₂， P， PRL， and FSH in serum （P<0.05， P<0.01）， along with a decreasing trend in LH level. In contrast， the BSZY groups at all doses exhibited significantly elevated levels of E₂， P， LH， PRL， and FSH in serum （P<0.05， P<0.01）. Network pharmacology predictions suggested that BSZY may exert protective effects against abortion in rats by activating the ERα/PI3K/Akt signaling pathway. Western blot results confirmed that BSZY significantly upregulated the expression of ERα， PI3K， and p-Akt proteins （P<0.05， P<0.01）. ConclusionBSZY has a protective effect on the abortion rats with kidney deficiency-corpus luteum inhibition syndrome， possibly by activating the ERα/PI3K/Akt signaling pathway to reduce ovarian apoptosis and regulate endocrine function， thereby lowering the abortion rate.

Objective:To explore the application strategies and clinical effects of superior gluteal artery perforator tissue flaps in repairing stage Ⅳ pressure ulcers in the sacrococcygeal region.Methods:This study was a retrospective observational study. From July 2019 to April 2024, 89 patients with stage Ⅳ pressure ulcers in the sacrococcygeal region who met the inclusion criteria were admitted to the First Affiliated Hospital of Nanchang University, including 59 males and 30 females, aged 21 to 84 years. There were 89 sacrococcygeal pressure ulcers, with an area of 5.0 cm×4.0 cm-21.0 cm×21.0 cm after debridement. According to the shape, size, and depth of the wounds after debridement, combined with the elasticity and texture of the skin around the wounds, and the principle of minimizing damage to the donor area, the appropriate forms of superior gluteal artery perforator tissue flaps were cut for wound repair in the following three conditions. (1) For wounds with a round shape, an area of 5.0 cm×5.0 cm-21.0 cm×21.0 cm, and a depth of 1.0-3.5 cm, the superior gluteal artery perforator propeller flap or myocutaneous flap, bilobed superior gluteal artery perforator relay flap, and bilateral superior gluteal artery perforator rotational flap were used. (2) For wounds with an oval shape, an area of 5.0 cm×4.0 cm-18.5 cm×10.5 cm, and a depth of 1.0-3.0 cm, the superior gluteal artery perforator propeller flap or myocutaneous flap, unilateral superior gluteal artery perforator propeller flap combined with contralateral superior gluteal artery perforator V-Y advanced flap or keystone flap were used. (3) For wounds with a fusiformis shape, an area of 7.0 cm×4.0 cm-17.5 cm×6.0 cm, and a depth of 1.5-5.0 cm, the unilateral or bilateral superior gluteal artery perforator V-Y advanced flap, superior gluteal artery perforator keystone flap, or superior gluteal artery perforator keystone flap combined with gluteus maximus muscle flap were used. In this group of patients, a total of 40 superior gluteal artery perforator propeller flaps (with an resection area of 11.0 cm×6.0 cm-17.0 cm×11.0 cm), 22 superior gluteal artery perforator propeller myocutaneous flaps (with an resection area of 10.0 cm×5.0 cm-14.0 cm×8.0 cm), 7 bilobed superior gluteal artery perforator relay flaps (with a main flap resection area of 5.5 cm×5.5 cm-18.0 cm×11.5 cm and a side flap resection area of 4.5 cm×3.0 cm-11.0 cm×6.5 cm), 5 bilateral superior gluteal artery perforator rotational flaps (with a total resection area of 20.0 cm×16.0 cm-26.0 cm×21.0 cm on both sides), 14 superior gluteal artery perforator V-Y advanced flaps (with an resection area of 12.0 cm×10.0 cm-18.0 cm×18.0 cm), 13 superior gluteal artery perforator keystone flaps (with an resection area of 13.0 cm×6.5 cm-19.0 cm×18.0 cm), and 3 gluteus maximus muscle flaps (with an resection area of 8.0 cm×3.0 cm-15.0 cm×4.5 cm). The donor area wounds were all directly sutured. The survival of tissue flaps was observed and the incidence rate of delayed wound healing in the reception area was calculated, and wound healing in the donor area was observed. The appearance and texture of tissue flaps and recurrence of pressure ulcers were followed up.Results:After surgery, all bilateral superior gluteal artery perforator rotational flaps, superior gluteal artery perforator V-Y advanced flaps, superior gluteal artery perforator keystone flaps, and gluteus maximus muscle flaps survived well. There were 6 cases of delayed wound healing in the reception area after surgery, with an incidence rate of 6.7% (6/89). Two patients had incision dehiscence in the donor area wounds due to postoperative bleeding, the wounds healed after debridement, vacuum sealing drainage, and dressing change. The wounds in the donor area of the remaining patients healed well. Six patients were lost to follow-up. Eighty-three patients were followed up for 3-48 months, of whom 4 patients died. Among the remaining 79 patients, 3 cases had pressure ulcers recur due to improper nursing, while the rest of the patients had tissue flaps with good appearance and soft texture and no recurrence of pressure ulcers.Conclusions:Based on the characteristics of wound shape, size, and depth after debridement of stage Ⅳ pressure ulcers in the sacrococcygeal region, individualized selection of flap, myocutaneous flap, or a combination of flap and gluteus maximus muscle flap based on the perforating branch of the superior gluteal artery perforator can achieve good clinical repair results. The postoperative tissue flap survived well, with a good appearance, soft texture, and less recurrence of pressure ulcers.

Objective:To explore the application strategies and clinical effects of superior gluteal artery perforator tissue flaps in repairing stage Ⅳ pressure ulcers in the sacrococcygeal region.Methods:This study was a retrospective observational study. From July 2019 to April 2024, 89 patients with stage Ⅳ pressure ulcers in the sacrococcygeal region who met the inclusion criteria were admitted to the First Affiliated Hospital of Nanchang University, including 59 males and 30 females, aged 21 to 84 years. There were 89 sacrococcygeal pressure ulcers, with an area of 5.0 cm×4.0 cm-21.0 cm×21.0 cm after debridement. According to the shape, size, and depth of the wounds after debridement, combined with the elasticity and texture of the skin around the wounds, and the principle of minimizing damage to the donor area, the appropriate forms of superior gluteal artery perforator tissue flaps were cut for wound repair in the following three conditions. (1) For wounds with a round shape, an area of 5.0 cm×5.0 cm-21.0 cm×21.0 cm, and a depth of 1.0-3.5 cm, the superior gluteal artery perforator propeller flap or myocutaneous flap, bilobed superior gluteal artery perforator relay flap, and bilateral superior gluteal artery perforator rotational flap were used. (2) For wounds with an oval shape, an area of 5.0 cm×4.0 cm-18.5 cm×10.5 cm, and a depth of 1.0-3.0 cm, the superior gluteal artery perforator propeller flap or myocutaneous flap, unilateral superior gluteal artery perforator propeller flap combined with contralateral superior gluteal artery perforator V-Y advanced flap or keystone flap were used. (3) For wounds with a fusiformis shape, an area of 7.0 cm×4.0 cm-17.5 cm×6.0 cm, and a depth of 1.5-5.0 cm, the unilateral or bilateral superior gluteal artery perforator V-Y advanced flap, superior gluteal artery perforator keystone flap, or superior gluteal artery perforator keystone flap combined with gluteus maximus muscle flap were used. In this group of patients, a total of 40 superior gluteal artery perforator propeller flaps (with an resection area of 11.0 cm×6.0 cm-17.0 cm×11.0 cm), 22 superior gluteal artery perforator propeller myocutaneous flaps (with an resection area of 10.0 cm×5.0 cm-14.0 cm×8.0 cm), 7 bilobed superior gluteal artery perforator relay flaps (with a main flap resection area of 5.5 cm×5.5 cm-18.0 cm×11.5 cm and a side flap resection area of 4.5 cm×3.0 cm-11.0 cm×6.5 cm), 5 bilateral superior gluteal artery perforator rotational flaps (with a total resection area of 20.0 cm×16.0 cm-26.0 cm×21.0 cm on both sides), 14 superior gluteal artery perforator V-Y advanced flaps (with an resection area of 12.0 cm×10.0 cm-18.0 cm×18.0 cm), 13 superior gluteal artery perforator keystone flaps (with an resection area of 13.0 cm×6.5 cm-19.0 cm×18.0 cm), and 3 gluteus maximus muscle flaps (with an resection area of 8.0 cm×3.0 cm-15.0 cm×4.5 cm). The donor area wounds were all directly sutured. The survival of tissue flaps was observed and the incidence rate of delayed wound healing in the reception area was calculated, and wound healing in the donor area was observed. The appearance and texture of tissue flaps and recurrence of pressure ulcers were followed up.Results:After surgery, all bilateral superior gluteal artery perforator rotational flaps, superior gluteal artery perforator V-Y advanced flaps, superior gluteal artery perforator keystone flaps, and gluteus maximus muscle flaps survived well. There were 6 cases of delayed wound healing in the reception area after surgery, with an incidence rate of 6.7% (6/89). Two patients had incision dehiscence in the donor area wounds due to postoperative bleeding, the wounds healed after debridement, vacuum sealing drainage, and dressing change. The wounds in the donor area of the remaining patients healed well. Six patients were lost to follow-up. Eighty-three patients were followed up for 3-48 months, of whom 4 patients died. Among the remaining 79 patients, 3 cases had pressure ulcers recur due to improper nursing, while the rest of the patients had tissue flaps with good appearance and soft texture and no recurrence of pressure ulcers.Conclusions:Based on the characteristics of wound shape, size, and depth after debridement of stage Ⅳ pressure ulcers in the sacrococcygeal region, individualized selection of flap, myocutaneous flap, or a combination of flap and gluteus maximus muscle flap based on the perforating branch of the superior gluteal artery perforator can achieve good clinical repair results. The postoperative tissue flap survived well, with a good appearance, soft texture, and less recurrence of pressure ulcers.

Objective To determine the humoral immunity in advanced maternal-age women with recurrent spontaneous abortion(RSA).Methods A retrospective study was performed between January 2022 and October 2023 in the Department of Reproductive Immunity of Shanghai First Maternity and Infant Hospital.Women with RSA were recruited and multiple autoantibodies were tested.Multivariate logistic regression was performed to compare the associations between different age groups(20 to 34 years old in the low maternal-age group and 35 to 45 years in the advanced maternal-age group)and multiple autoantibodies,while controlling for three confounding factors,including body mass index(BMI),previous history of live birth,and the number of spontaneous abortions.Then,we investigated the differences in the humoral immunity of advanced maternal-age RSA women and low maternal-age RSA women.Result A total of 4 009 women with RSA were covered in the study.Among them,1 158 women were in the advanced maternal-age group and 2 851 women were in the low maternal-age group.The prevalence of antiphospholipid syndrome,systemic lupus erythematosus,Sjogren's syndrome,rheumatoid arthritis,and undifferentiated connective tissue disease was 15.6% and 14.1%,0.0% and 0.1%,0.9% and 0.9%,0.3% and 0.0%,and 23.7% and 22.6% in the advanced maternal-age group and low maternal-age group,respectively,showing no statistical difference between the two groups.The positive rates of antiphospholipid antibodies(aPLs),antinuclear antibody(ANA),extractable nuclear antigen(ENA)antibody,anti-double stranded DNA(dsDNA)antibody,anti single-stranded DNA(ssDAN)antibody,antibodies against alpha-fodrin(AAA),and thyroid autoimmunity(TAI)were 19.1% and 19.5%,6.6% and 6.6%,9.2% and 10.5%,2.0% and 2.0%,2.2% and 1.2%,5.1% and 4.9%,and 17.8% and 16.8%,respectively.No differences were observed between the two groups.1.6% of the women in the advanced maternal-age group tested positive for lupus anticoagulant(LA),while 2.7% of the women in the low maternal-age group were LA positive,with the differences being statistically significant(odds ratio=0.36,95% confidence interval:0.17-0.78).In the 4 008 RSA patients,the cumulative cases tested positive for the three antibodies of the aPLs spectrum were 778,of which 520 cases were positive for anti-β2 glycoprotein Ⅰ antibodies(β2GPⅠ Ab)-IgG/IgM,58 were positive for aCL-IgG/IgM,73 were positive for LA,105 were positive for both β2GPⅠAb-IgG/IgM and aCL-IgG/IgM,17 were positive for both β2GPⅠ Ab-IgG/IgM and LA,2 were positive for both aCL-IgG/IgM and LA,and 3 were positive for all three antibodies.Conclusion Our study did not find a difference in humoral immunity between RSA women of advanced maternal age and those of low maternal age.

Objective:To explore the clinical efficacy of posterior femoral cutaneous nerve nutrient vessel adipofascial flaps plus free-style gluteal perforator flaps in repairing stage IV sciatic tuberosity pressure ulcers.Methods:A retrospective case series study was conducted to analyze the clinical data of 16 patients (16 wounds) with stage IV sciatic tuberosity pressure ulcers admitted to First Affiliated Hospital of Nanchang University from May 2021 to February 2024, including 10 males and 6 females, aged 21-84 years [(58.5±16.5)years]. Among them, 8 patients were complicated with chronic osteomyelitis of the ischium at 8 sites. The wound area before debridement ranged from 2.0 cm×1.5 cm to 9.0 cm×7.0 cm. All the patients underwent staged surgery. In phase I surgery, the scar tissue at the wound margin, necrotic tissue, bursa, and chronic osteomyelitic lesions were removed in the ischium. After debridement, the wound area ranged from 4.0 cm×3.0 cm to 12.0 cm×8.0 cm. Negative pressure closure drainage (VSD) was performed and wound bed preparation was completed. In phase II surgery, the posterior femoral cutaneous nerve nutrient vessel adipofascial flaps were flipped, filled into the wound cavity, and then used to repair the wound by advancing and rotating in combination with free-style gluteal perforator flap. The area of posterior femoral cutaneous nerve nutrient vessel adipofascial flaps ranged from 9.0 cm×3.5 cm to 19.0 cm×10.0 cm and the area of the free-style gluteal perforator flaps ranged from 5.0 cm×4.0 cm to 13.0 cm×8.5 cm. The amount of bleeding in phase II surgery was recorded. The survival and wound healing of the posterior femoral cutaneous nerve nutrient vessel adipofascial flaps and free-style gluteal perforator flaps were observed. At the last follow-up, recurrence of pressure ulcers and osteomyelitis, external appearance of the wound, and secondary functional impairment and deformity in the donor sites were observed.Results:All the patients were followed up for 6-15 months [(9.4±3.1)months]. The intraoperative bleeding volume in phase II surgery was 80-300 ml [(162.9±60.6)ml]. All the posterior femoral cutaneous nerve nutrient vessel adipofascial flaps survived well after surgery. A small area of bruising was observed at the distal end of the freestyle gluteal perforator flap in 1 patient at 1 day after surgery, which was relieved after removing some of the sutures. Torn suture of the incision was found as a result of postoperative subcutaneous hematoma in the donor site of the posterior femoral cutaneous nerve nutrient vessel adipofascial flap in 1 patient at 1 day after surgery, which healed at 22 days after bedside debridement and dressing change. All other incisions healed well. At the last follow-up, there was no recurrence of pressure ulcers or osteomyelitis and the wound was mildly pigmented and soft. There were no secondary functional impairments or deformities in the posterior femoral or gluteal donor sites.Conclusion:Posterior femoral cutaneous nerve nutrient vessel adipofascial flaps plus freestyle gluteal perforator flaps can be used in the repair of stage IV sciatic tuberosity pressure ulcer wounds, with the advantages of less intraoperative bleeding, high tissue flap survival rate, good wound healing, no recurrence of pressure ulcers or osteomyelitis after surgery, good wound appearance and texture, and no secondary functional impairment or deformity in the donor sites.

Objective:To explore the clinical efficacy of posterior femoral cutaneous nerve nutrient vessel adipofascial flaps plus free-style gluteal perforator flaps in repairing stage IV sciatic tuberosity pressure ulcers.Methods:A retrospective case series study was conducted to analyze the clinical data of 16 patients (16 wounds) with stage IV sciatic tuberosity pressure ulcers admitted to First Affiliated Hospital of Nanchang University from May 2021 to February 2024, including 10 males and 6 females, aged 21-84 years [(58.5±16.5)years]. Among them, 8 patients were complicated with chronic osteomyelitis of the ischium at 8 sites. The wound area before debridement ranged from 2.0 cm×1.5 cm to 9.0 cm×7.0 cm. All the patients underwent staged surgery. In phase I surgery, the scar tissue at the wound margin, necrotic tissue, bursa, and chronic osteomyelitic lesions were removed in the ischium. After debridement, the wound area ranged from 4.0 cm×3.0 cm to 12.0 cm×8.0 cm. Negative pressure closure drainage (VSD) was performed and wound bed preparation was completed. In phase II surgery, the posterior femoral cutaneous nerve nutrient vessel adipofascial flaps were flipped, filled into the wound cavity, and then used to repair the wound by advancing and rotating in combination with free-style gluteal perforator flap. The area of posterior femoral cutaneous nerve nutrient vessel adipofascial flaps ranged from 9.0 cm×3.5 cm to 19.0 cm×10.0 cm and the area of the free-style gluteal perforator flaps ranged from 5.0 cm×4.0 cm to 13.0 cm×8.5 cm. The amount of bleeding in phase II surgery was recorded. The survival and wound healing of the posterior femoral cutaneous nerve nutrient vessel adipofascial flaps and free-style gluteal perforator flaps were observed. At the last follow-up, recurrence of pressure ulcers and osteomyelitis, external appearance of the wound, and secondary functional impairment and deformity in the donor sites were observed.Results:All the patients were followed up for 6-15 months [(9.4±3.1)months]. The intraoperative bleeding volume in phase II surgery was 80-300 ml [(162.9±60.6)ml]. All the posterior femoral cutaneous nerve nutrient vessel adipofascial flaps survived well after surgery. A small area of bruising was observed at the distal end of the freestyle gluteal perforator flap in 1 patient at 1 day after surgery, which was relieved after removing some of the sutures. Torn suture of the incision was found as a result of postoperative subcutaneous hematoma in the donor site of the posterior femoral cutaneous nerve nutrient vessel adipofascial flap in 1 patient at 1 day after surgery, which healed at 22 days after bedside debridement and dressing change. All other incisions healed well. At the last follow-up, there was no recurrence of pressure ulcers or osteomyelitis and the wound was mildly pigmented and soft. There were no secondary functional impairments or deformities in the posterior femoral or gluteal donor sites.Conclusion:Posterior femoral cutaneous nerve nutrient vessel adipofascial flaps plus freestyle gluteal perforator flaps can be used in the repair of stage IV sciatic tuberosity pressure ulcer wounds, with the advantages of less intraoperative bleeding, high tissue flap survival rate, good wound healing, no recurrence of pressure ulcers or osteomyelitis after surgery, good wound appearance and texture, and no secondary functional impairment or deformity in the donor sites.

ObjectiveTo investigate the effect and mechanism of osthole on the proliferation and apoptosis in human intrahepatic cholangiocarcinoma HuCCT1 cells. MethodThe effect of 10， 20， 40， 80， and 120 μmol·L^-1 osthole on the proliferation of HuCCT1 cells was detected by the cell counting kit-8 （CCK-8）. A blank group， and low-， medium-， and high-dose osthole groups （16， 32， and 64 μmol·L^-1） were set up. The effect of osthole on cell clone formation rate was detected by colony formation assay. The effect of osthole on cell cycle and apoptosis was detected by flow cytometry. The effect of osthole on cell apoptotic morphology was detected by Hoechst 33342 fluorescent staining. The effect of osthole on cell cycle protein cyclin B₁， proliferating cell nuclear antigen （PCNA）， cysteine-aspartic acid protease （Caspase）-9， Caspase-3， cleaved Caspase-9， cleaved Caspase-3， cleaved poly（ADP-ribose） polymerase （cleaved PARP）， B-cell lymphoma-2 （Bcl-2）， phosphorylated protein kinase B （p-Akt）， phosphorylated mammalian target of rapamycin （p-mTOR）， and phosphorylated ribosomal protein S6 （p-RPS6） was detected by Western blot. ResultThe cell viability in the osthole group（40，80，120 μmol·L^-1） decreased （P<0.05，P<0.01）， with the half maximal inhibitory concentration （IC₅₀） of 63.8 μmol·L^-1 as compared with that in the blank group. Compared with the blank group， the osthole groups（32，64 μmol·L^-1）showed reduced clone formation rate （P<0.01）， increased number of cells in the G₂ phase （P<0.05，P<0.01）， decreased number of cells， increased pyknosis and fragmentation， increased apoptosis rate （P<0.05，P<0.01）， down-regulated expression of cyclin B₁， PCNA， Bcl-2， Caspase-3， Caspase-9， p-Akt， p-mTOR， and p-RPS6 （P<0.05，P<0.01）， and up-regulated expression of cleaved Caspase-3， cleaved Caspase-9， and cleaved PARP （P<0.05，P<0.01）. ConclusionOsthole can inhibit the proliferation and promote the apoptosis of HuCCT1 cells， and its mechanism may be related to the Akt/mTOR signaling pathway.

The aim of this study was to refold the OvisAries leukocyte antigen (OLA) class Ⅰ protein with peptides derived from sheeppox virus (SPPV) to identify SPPV T cell epitopes. Two pairs of primers were designed based on the published sequence of a sheep major histocompatibility complex Ⅰ to amplify the heavy chain gene of OLA Ⅰ α-BSP and the light chain gene of OLA Ⅰ-β₂m. Both genes were cloned into a pET-28a(+) expression vector, respectively, and induced with ITPG for protein expression. After purification, the heavy chain and light chain proteins as well as peptides derived from SPPV were refolded at a ratio of 1:1:1 using a gradual dilution method. Molecular exclusion chromatography was used to test whether these peptides bind to the OLA Ⅰ complex. T-cell responses were assessed using freshly isolated PBMCs from immunized sheep through IFN-γ ELISPOT with peptides derived from SPPV protein. The results showed that the cloned heavy chain and light chain expressed sufficiently, with a molecular weight of 36.3 kDa and 16.7 kDa, respectively. The protein separated via a Superdex^TM 200 increase 10/300 GL column was collected and verified by SDS-PAGE after refolding. One SPPV CTL epitope was identified after combined refolding and functional studies based on T-cell epitopes derived from SPPV. An OLA Ⅰ/peptide complex was refolded correctly, which is necessary for the structural characterization. This study may contribute to the development of sheep vaccine based on peptides.
Animals ; Capripoxvirus ; Epitopes, T-Lymphocyte/genetics* ; Peptides/genetics* ; Poxviridae Infections ; Sheep ; Sheep Diseases

ObjectiveTo explore the mechanism of cucurbitacin B （CuB） in inhibiting cell proliferation and glycolysis. MethodCell counting kit-8 （CCK-8） was applied to investigate the effect of different concentrations of CuB （0， 40， 80， 120， 160， 200， 400， and 800 nmol·L^-1） on the proliferation of HuCCT1 cells. The effect of different concentrations of CuB （50， 100， and 200 nmol·L^-1） on the colony formation ability of HuCCT1 cells was detected by plate cloning assay. The effect of different concentrations of CuB （50， 100， 200 nmol·L^-1） on the HuCCT1 cell cycle was analyzed by flow cytometry. Visible spectrophotometry was employed to detect the activity of key glycolytic enzymes hexokinase （HK） and pyruvate kinase （PK）） and changes in glucose consumption， lactate production， and adenosine triphosphate （ATP） production in HuCCT1 cells after administration of different concentrations of CuB （50， 100， 200 nmol·L^-1）. Western blotting was used to assay the effect of CuB on the expression of cell cycle-related proteins， proliferation-related proteins， key glycolytic proteins， and Akt/mammalian target of rapamycin （mTOR） pathway-related proteins. ResultAs compared with the blank group， CuB at dose of 160-800 nmol·L^-1 after 24 h administration and CuB at dose of 80-800 nmol·L^-1 after 48 h administration inhibited the proliferation of HuCCT1 cells in a time- and dose-dependent manner （P<0.05， P<0.01）， and the median inhibitory concentration was 200 nmol·L^-1 48 h after administration. CuB can restrain the colony formation ability of HuCCT1 cells in a dose-dependent manner （P<0.01）， and block HuCCT1 cell cycle in G₂ phase （P<0.05， P<0.01）. CuB （100 and 200 nmol·L^-1） can suppress the activities of HK and PK and reduce cell glucose consumption and production of lactate and ATP （P<0.05， P<0.01）. Western blot results showed that CuB （100 and 200 nmol·L^-1） can inhibit the protein levels of cycle-related protein Cyclin B₁， proliferating cell nuclear antigen （PCNA）， HK1， HK2， PKM1， PKM2， phosphorylated Akt （p-Akt）， phosphorylated mTOR （p-mTOR）， and phosphorylated ribosomal protein S6 （p-RPS6） （P<0.05， P<0.01）. ConclusionCuB can inhibit aerobic glycolysis in HuCCT1 cells via the Akt/mTOR pathway， thereby affecting cell proliferation.

Objective:To compare the efficacy of morphine with intravenous injection and subcutaneous injection in the treatment of advanced cancer,and explore the indications of different drug delivery methods for high-dose morphine. Methods: A prospective study was performed,and 46 cases of patients with advanced cancer pain were collected and divided into intravenous group and subcutaneous group according to the administration route. Pain was assessed during the administration,and the analgesic efficiency and the incidence of adverse reactions were observed to compare the efficacy and safety of two different ways to give high-dose morphine. Results:No statisti-cally significant differences were found in the number of outbreaks needed to be rescued,the frequency of morphine-induced drug deliver-y,the efficiency of analgesia after opioid transfer,and the incidence of opioid-related side effects between the groups (P>0.05). The dose of morphine in the subcutaneous group was higher than that of the intravenous group(P<0.05). Conclusion:The continuous ad-ministration of morphine with intravenous injection and subcutaneous injection can quickly,safely and effectively relieve pain. With the same analgesic efficacy,patients can choose appropriate administration route according to the dose of morphine, the influence degree of primary diseases and the individual will.