Objective To analyze the trends in the disease burden of schistosomiasis in China from 1992 to 2021, and to project the disease burden of schistosomiasis in China from 2022 to 2030, so as to provide insights into the elimination of schistosomiasis in China. Methods The prevalence, age-standardized prevalence, disability-adjusted life year (DALYs) rate and age-standardized DALYs rate of schistosomiasis, as well as the years lost due to disability (YLDs) rate and age-standardized YLDs rate of anemia attributable to Schistosoma infections in China, the world and different socio-demographic index (SDI) regions were captured from the Global Burden of Disease Study 2021 (GBD 2021) data resources, and the trends in the disease burden due to schistosomiasis were evaluated with estimated annual percentage change (EAPC) and its 95% confidence interval (CI). In addition, the age, period and cohort effects on the prevalence of schistosomiasis were examined in China using an age-period-cohort (APC) model, and the disease burden of schistosomiasis was predicted in China from 2022 to 2030 using a Bayesian age-period-cohort (BAPC) model. Results The age-standardized prevalence and DALYs rate of schistosomiasis, and the age-standardized YLDs rate of anemia attributable to Schistosoma infections were 761.32/10⁵, 5.55/10⁵ and 0.38/10⁵ in China in 2021. These rates were all lower than the global levels (1 914.30/10⁵, 21.90/10⁵ and 3.36/10⁵, respectively), as well as those in the medium SDI regions (1 413.61/10⁵, 12.10/10⁵ and 1.93/10⁵, respectively), low-medium SDI regions (2 461.03/10⁵, 26.81/10⁵ and 4.48/10⁵, respectively), and low SDI regions (5 832.77/10⁵, 94.48/10⁵ and 10.65/10⁵, respectively), but higher than those in the high SDI regions (59.47/10⁵, 0.49/10⁵ and 0.05/10⁵, respectively) and high-medium SDI regions (123.11/10⁵, 1.20/10⁵ and 0.12/10⁵, respectively). The prevalence and DALYs rate of schistosomiasis were higher among men (820.79/10⁵ and 5.86/10⁵, respectively) than among women (697.96/10⁵ and 5.23/10⁵, respectively) in China in 2021, while the YLDs rate of anemia attributable to Schistosoma infections was higher among women (0.66/10⁵) than among men (0.12/10⁵). The prevalence of schistosomiasis peaked at ages of 30 to 34 years among both men and women, while the DALYs rate of schistosomiasis peaked among men at ages of 15 to 19 years and among women at ages of 20 to 24 years. The age-standardized prevalence of schistosomiasis showed a moderate decline in China from 1992 to 2021 relative to different SDI regions [EAPC = -1.51%, 95% CI: (-1.65%, -1.38%)], while the age-standardized DALYs rate [EAPC = -3.61%, 95% CI: (-3.90%, -3.33%)] and age-standardized YLDs rate of anemia attributable to Schistosoma infections [EAPC = -4.16%, 95% CI: (-4.38%, -3.94%)] appeared the fastest decline in China from1992 to 2021 relative to different SDI regions. APC modeling showed age, period, and cohort effects on the trends in the prevalence of schistosomiasis in China from 1992 to 2021, and the prevalence of schistosomiasis appeared a rise followed by decline with age, and reduced with period and cohort. BAPC modeling revealed that the age-standardized prevalence and age-standardized DALYs rate of schistosomiasis, and age-standardized YLDs rate of anemia attributable to Schistosoma infections all appeared a tendency towards a decline in China from 2022 to 2030, which reduced to 722.72/10⁵ [95% CI: (538.74/10⁵, 906.68/10⁵)], 5.19/10⁵ [95% CI: (3.54/10⁵, 6.84/10⁵)] and 0.30/10⁵ [95% CI: (0.21/10⁵, 0.39/10⁵)] in 2030, respectively. Conclusions The disease burden of schistosomiasis appeared a tendency towards a decline in China from 1992 to 2021, and is projected to appear a tendency towards a decline from 2022 to 2030. There are age, period and cohort effects on the prevalence of schistosomiasis in China. Precision schistosomiasis control is required with adaptations to current prevalence and elimination needs.

This study aims to establish an indirect ELISA method for detecting RVFV antibodies u-sing recombinant proteins of Rift Valley fever virus(RVFV)Gn protein Ⅱ-Ⅲ structural domains as the encapsulated antigen which was expressed by the Escherichia coli(E.coli)expression sys-tem.The gene sequences encoding the Ⅱ and Ⅲ subdomains of RVFV Gn protein were inserted in-to pET-30a(+)to construct the recombinant plasmid pET-RVFV Gn-D Ⅱ-Ⅲ.After transforma-tion of the recombinant plasmid into DE3(BL21)competent cells,the recombinant Gn-D Ⅱ-Ⅲ protein was induced with IPTG and purified using affinity chromatography.An indirect ELISA method for the detection of RVFV antibodies was developed using purified recombinant protein as coating antigen and SPA-HRP as the enzyme-labelled secondary antibody.Western blot analysis confirmed that the RVFV Gn-D Ⅱ-Ⅲ protein was successfully expressed.The optimal expression conditions for RVFV Gn-D Ⅱ-Ⅲ protein were induced with 0.8 mmol/L IPTG at 37 ℃ for 5 h.The Gn-D Ⅱ-Ⅲ protein was purified using affinity chromatography with a purity of 91.9％,and the purified protein was used as the encapsulated antigen to develop an ELISA assay for RVFV anti-bodies.The specificity evaluation showed that the method specifically detected RVFV-positive sera and did not cross-react with sera positive for West Nile virus(WNV),Ebola virus(EBOV),Mar-burg virus(MARV)and tick-borne encephalitis virus(TBEV).When the RVFV Gn-D Ⅲ-Ⅲ posi-tive serum was diluted to 6 400 times,the test result still showed positive results,demonstrating the method had good sensitivity.The repeatability evaluation results indicated that the variation co-efficients for both intra-and inter-batch responses was less than 10％,indicating that the method had good repeatability.In conclusion,the RVFV Gn-D Ⅱ-Ⅲ protein was successfully expressed u-sing the E.coli expression system.The purified recombinant Gn-D Ⅱ-Ⅲ protein was used as the encapsulated antigen to develop an indirect ELISA assay for RVFV antibodies,which provides a preliminary basis for the diagnosis of RVF and the research and development of RVF vaccines.

This study aims to establish an indirect ELISA method for detecting RVFV antibodies u-sing recombinant proteins of Rift Valley fever virus(RVFV)Gn protein Ⅱ-Ⅲ structural domains as the encapsulated antigen which was expressed by the Escherichia coli(E.coli)expression sys-tem.The gene sequences encoding the Ⅱ and Ⅲ subdomains of RVFV Gn protein were inserted in-to pET-30a(+)to construct the recombinant plasmid pET-RVFV Gn-D Ⅱ-Ⅲ.After transforma-tion of the recombinant plasmid into DE3(BL21)competent cells,the recombinant Gn-D Ⅱ-Ⅲ protein was induced with IPTG and purified using affinity chromatography.An indirect ELISA method for the detection of RVFV antibodies was developed using purified recombinant protein as coating antigen and SPA-HRP as the enzyme-labelled secondary antibody.Western blot analysis confirmed that the RVFV Gn-D Ⅱ-Ⅲ protein was successfully expressed.The optimal expression conditions for RVFV Gn-D Ⅱ-Ⅲ protein were induced with 0.8 mmol/L IPTG at 37 ℃ for 5 h.The Gn-D Ⅱ-Ⅲ protein was purified using affinity chromatography with a purity of 91.9％,and the purified protein was used as the encapsulated antigen to develop an ELISA assay for RVFV anti-bodies.The specificity evaluation showed that the method specifically detected RVFV-positive sera and did not cross-react with sera positive for West Nile virus(WNV),Ebola virus(EBOV),Mar-burg virus(MARV)and tick-borne encephalitis virus(TBEV).When the RVFV Gn-D Ⅲ-Ⅲ posi-tive serum was diluted to 6 400 times,the test result still showed positive results,demonstrating the method had good sensitivity.The repeatability evaluation results indicated that the variation co-efficients for both intra-and inter-batch responses was less than 10％,indicating that the method had good repeatability.In conclusion,the RVFV Gn-D Ⅱ-Ⅲ protein was successfully expressed u-sing the E.coli expression system.The purified recombinant Gn-D Ⅱ-Ⅲ protein was used as the encapsulated antigen to develop an indirect ELISA assay for RVFV antibodies,which provides a preliminary basis for the diagnosis of RVF and the research and development of RVF vaccines.

Feline herpesvirus type Ⅰ(FHV-1)was used as the vector.The gI and gE genes of FHV-1 were replaced with the feline calicivirus(FCV)VP1 gene and the red fluorescent protein(mCherry)gene by CRISPR/Cas9 systems and homologous recombination technology,and the re-combinant virus strain FHV △gI&gE/VP1-mCherry+was successfully rescued.The recombinant virus strain was purified by plaque assay.The biological characteristics and genetic stability of the recombinant virus were analyzed by indirect immunofluorescence assay,plaque morphological anal-ysis,and PCR.The results of the indirect immunofluorescence identification showed that the re-combinant virus FHV △gI&gE/VP1-mCherry+could express the VP1 protein in F81 cells,and the growth characteristics of the recombinant virus were not significantly different from those of the parent virus FHV-1.The plaque morphology and staining results indicated that the area of the plaque formed by the recombinant virus was smaller than that of the parent virus,suggesting that the spread ability of the recombinant virus between cells was reduced after the deletion of the gI and gE genes.The result of PCR showed that the VP1 gene could still be detected after 15 succes-sive passages of the recombinant virus,indicating that the recombinant virus had good genetic stability.In this study,the recombinant virus strain expressing the FCV VP1 protein was successfully prepared,which will lay a foundation for the development of engineered FCV and FHV-1 vaccine.

Feline herpesvirus type Ⅰ(FHV-1)was used as the vector.The gI and gE genes of FHV-1 were replaced with the feline calicivirus(FCV)VP1 gene and the red fluorescent protein(mCherry)gene by CRISPR/Cas9 systems and homologous recombination technology,and the re-combinant virus strain FHV △gI&gE/VP1-mCherry+was successfully rescued.The recombinant virus strain was purified by plaque assay.The biological characteristics and genetic stability of the recombinant virus were analyzed by indirect immunofluorescence assay,plaque morphological anal-ysis,and PCR.The results of the indirect immunofluorescence identification showed that the re-combinant virus FHV △gI&gE/VP1-mCherry+could express the VP1 protein in F81 cells,and the growth characteristics of the recombinant virus were not significantly different from those of the parent virus FHV-1.The plaque morphology and staining results indicated that the area of the plaque formed by the recombinant virus was smaller than that of the parent virus,suggesting that the spread ability of the recombinant virus between cells was reduced after the deletion of the gI and gE genes.The result of PCR showed that the VP1 gene could still be detected after 15 succes-sive passages of the recombinant virus,indicating that the recombinant virus had good genetic stability.In this study,the recombinant virus strain expressing the FCV VP1 protein was successfully prepared,which will lay a foundation for the development of engineered FCV and FHV-1 vaccine.

Objective To study changes in immune cells and cytokines during the reactivation stage of varicella-zoster virus(VZV)in patients with herpes zoster.Methods A total of 50 patients with herpes zoster and 30 healthy individuals were selected from Xi'an Ninth Hospital between May 2022 and October 2022.Flow cytometry was used to detect the proportion of peripheral blood CD3+cells,CD4+cells,CD8+T cells,B cells and NK cells,as well as levels of cytokines IL-2,IFN-γ,IL-10 and IL-6.We analyzed the immune mechanism of VZV reactivation stage in herpes zoster patients.Results Compared with the healthy control group,the proportion of CD3+cells and CD4+T cells in herpes zoster patients decreased significantly;the proportion of NK cells significantly increased;the levels of IFN-γ,IL-10 and IL-6 significantly increased;the proportion of CD8+T cells,B cells and IL-2 content showed an increasing trend,but there was no significant difference.In addition,the severity of neurological involvement in herpes zoster patients might affect changes in cytokine levels.Conclusion During the reactivation period of VZV,changes in the proportion of immune cells and cytokine expression levels are closely related to the occurrence and development of herpes zoster.

BACKGROUND:As a member of bone morphogenetic proteins,growth differentiation factor-5 shows promising potential in the application of cartilage and bone repair.The affinity of growth differentiation factor-5 onto bone tissue determines protein use efficiency,so it is of great significance to prepare growth differentiation factor-5 with bone targeting capability. OBJECTIVE:To modify growth differentiation factor-5 using bisphosphonates and investigate the effects of modified protein on the growth of preosteoblasts in mice. METHODS:Pamidronate disodium/growth differentiation factor-5 complex was prepared using chemical crosslinking to couple growth differentiation factor-5 with pamidronate disodium.The functional groups and structures of the complex were characterized using Fourier transform infrared spectroscopy and circular dichromatography.To determine the bone targeting in vitro,the binding of the modified growth differentiation factor-5 with calcium phosphate and in vitro release amount of growth differentiation factor-5 were measured with an ELISA kit.Growth differentiation factor-5(control group)and the pamidronate disodium/growth differentiation factor-5 complex(experimental group)were co-cultured with preosteoblasts MC3T3-E1.Individually cultured cells were blank controls.The effect of the complex on cell proliferation and differentiation was evaluated. RESULTS AND CONCLUSION:(1)The infrared spectroscopy and circular dichromatography results indicated that the bisphosphonate/growth differentiation factor-5 complex was successfully prepared without significant changes in the protein secondary structure.In vitro protein adsorption results showed that growth differentiation factor-5 adsorption on calcium phosphate was increased by about one time after coupling with a bisphosphonate.In the presence of cysteine,growth differentiation factor-5 could be released from the bisphosphonate/growth differentiation factor-5 complex.(2)CCK-8 assay results showed that the absorbance value of the experimental group cultured for 4 and 7 days was higher than that of the control group and blank control group(P<0.000 1).After 7 days of culture,the expression of alkaline phosphatase in the experimental group was significantly higher than that in the control group and blank control group(P<0.000 1).After 13 days of culture,the content of calcium nodules in the experimental group was significantly higher than that in the control group and the blank control group(P<0.000 1).The results of qRT-PCR showed that the mRNA expression of alkaline phosphatase,osteocalcin and Runx2 in the experimental group was higher than that in the control group and the blank control group after 7 days of culture(P<0.01,P<0.001,P<0.000 1).(3)These findings exhibit that bisphosphonate modification can enhance the binding capacity of growth differentiation factor-5 to calcium phosphate as well as improve its biological activity.

Background::Pancreatic ductal adenocarcinoma (PDAC) is one of the main types of malignant tumor of the digestive system, and patient prognosis is affected by difficulties in early diagnosis, poor treatment response, and a high postoperative recurrence rate. Carbohydrate antigen 19-9 (CA19-9) has been widely used as a biomarker for the diagnosis and postoperative follow-up of PDAC patients. Nevertheless, the production mechanism and potential role of CA19-9 in PDAC progression have not yet been elucidated.Methods::We performed single-cell RNA sequencing on six samples pathologically diagnosed as PDAC (three CA19-9-positive and three CA19-9-negative PDAC samples) and two paracarcinoma samples. We also downloaded and integrated PDAC samples (each from three CA19-9-positive and CA19-9-negative patients) from an online database. The dynamics of the proportion and potential function of each cell type were verified through immunofluorescence. Moreover, we built an in vitro coculture cellular model to confirm the potential function of CA19-9. Results::Three subtypes of cancer cells with a high ability to produce CA19-9 were identified by the markers TOP2A, AQP5, and MUC5AC. CA19-9 production bypass was discovered on antigen-presenting cancer-associated fibroblasts (apCAFs). Importantly, the proportion of immature ficolin-1 positive (FCN1+) macrophages was high in the CA19-9-negative group, and the proportion of mature M2-like macrophages was high in the CA19-9-positive group. High proportions of these two macrophage subtypes were associated with an unfavourable clinical prognosis. Further experiments indicated that CA19-9 could facilitate the transformation of M0 macrophages into M2 macrophages in the tumor microenvironment. Conclusions::Our study described CA19-9 production at single-cell resolution and the dynamics of the immune atlas in CA19-9-positive and CA19-9-negative PDAC. CA19-9 could promote M2 polarization of macrophage in the pancreatic tumor microenvironment.

Objective:To compare the effects of pressure controlled ventilation (PCV) and volume controlled ventilation (VCV) on respiratory mechanics, hemodynamics and biochemical metabolism in patients undergoing laparoscopic colorectal cancer surgery.Methods:The clinical data of 78 patients underwent laparoscopic colorectal cancer surgery from August 2019 to June 2020 in Inner Mongolia People′s Hospital were retrospectively analyzed. Among them, 39 patients were treated with PCV (PCV group), and 39 were treated with VCV (VCV group). The respiratory mechanics, hemodynamics and biochemical metabolism indexes 10 min after anesthesia induction (T 1), 10 min after pneumoperitoneum + low head and foot height (T 2), 60 min after pneumoperitoneum + low head and foot height (T 3) and 120 min after pneumoperitoneum + low head and foot height (T 4) were recorded. The respiratory mechanical indexes included mean airway pressure (P mean), airway peak pressure (P peak), pressure of end tidal carbon dioxide (P ETCO 2) and dynamic lung compliance (C Ldyn); hemodynamic indexes included mean arterial pressure (MAP) and heart rate; and biochemical metabolic indexes included base excess, serum natrium, serum potassium, negative logarithm of the hydrogen ion concentration (pH) and blood glucose. Results:The P mean, P peak and P ETCO 2 T 1 to T 4 in PCV group were significantly lower than those in VCV group, P mean: (7.12 ± 1.37) cmH 2O (1 cmH 2O = 0.098 kPa) vs. (8.54 ± 1.84) cmH 2O, (9.80 ± 2.26) cmH 2O vs. (11.63 ± 2.87) cmH 2O, (9.51 ± 2.17) cmH 2O vs. (11.72 ± 2.90) cmH 2O, (7.04 ± 1.34) cmH 2O vs. (8.65 ± 1.88) cmH 2O; P peak: (13.41 ± 2.68) cmH 2O vs. (15.06 ± 3.05) cmH 2O, (20.92 ± 3.11) cmH 2O vs. (23.45 ± 4.02) cmH 2O, (21.14 ± 3.50) cmH 2O vs. (23.69 ± 4.26) cmH 2O, (15.03 ± 2.74) cmH 2O vs. (16.45 ± 3.21) cmH 2O; P ETCO 2: (30.59 ± 1.57) mmHg (1 mmHg = 0.133 kPa) vs. (32.04 ± 2.11) mmHg, (35.02 ± 4.15) mmHg vs. (39.88 ± 4.76) mmHg, (35.90 ± 4.22) mmHg vs. (40.11 ± 4.87) mmHg, (34.33 ± 4.17) mmHg vs. (37.65 ± 2.69) mmHg; the C Ldyn was significantly higher than that in VCV group: (40.68 ± 3.98) ml/cmH 2O vs. (35.47 ± 2.56) ml/cmH 2O, (30.25 ± 3.21) ml/cmH 2O vs. (22.40 ± 2.75) ml/cmH 2O, (29.78 ± 3.06) ml/cmH 2O vs. (22.60 ± 2.81) ml/cmH 2O, (40.32 ± 4.25) ml/cmH 2O vs. (33.61 ± 2.81) ml/cmH 2O, and there were statistical differences ( P<0.01 or <0.05). The MAP and heart rate T 1 to T 4 in PCV group were significantly lower than those in VCV group, and there was statistical difference ( P<0.01 or <0.05). The base excess, serum natrium, serum potassium and pH T 1 to T 4 in PCV group were significantly higher than those in VCV group, while the blood glucose was significantly lower than that in VCV group, and there was statistical difference ( P<0.01 or <0.05). Conclusions:Compared with VCV, PCV can promote the recovery of respiratory mechanics index, stabilize hemodynamics and improve biochemical metabolism index in patients undergoing laparoscopic colorectal cancer surgery.

Objective:To evaluate the role of silent information regulator 1 (SIRT1)/nuclear factor E2 related factor 2 (Nrf2) signaling pathway in sleep deprivation-induced cognitive dysfunction in rats.Methods:Thirty-six adult male Sprague-Dawley rats, aged 54-56 weeks, weighing 600-750 g, were divided into 3 groups ( n=12 each) using a random number table method: control group (group C), sleep deprivation group (group SD) and sleep deprivation+ SIRT1 agonist Srt1720 group (group SD+ Srt). Sleep deprivation model was established by modified multi-platform water environment method.Srt1720 10 mg/kg was injected intraperitoneally every 12 h starting from 24 h before establishing the model in group SD+ Srt, while the equal volume of 0.9% normal saline was injected intraperitoneally in C and SD groups.After the end of sleep deprivation, Morris water maze test was performed to evaluate the cognitive function.Animals were then sacrificed, and their hippocampi were removed for determination of neuronal degeneration rate in hippocampal CA1 region (using HE staining), the apoptosis rate in hippocampal CA1 (using TUNEL assay ), the expression of SIRT1 and Nrf2 in hippocampal CA1 (by immunohistochemistry) and the contents of reactive oxygen species (ROS) and superoxide dismutase (SOD) (by microplate method). Results:Compared with group C, the escape latency was significantly prolonged, the time of staying at the platform quadrant was shortened, and the frequency of crossing the platform was decreased on 2-4 days, the apoptotic rate and neuronal degeneration rate were increased, the expression of SIRT1 and Nrf2 was down-regulated, ROC content was increased, and SOD content was decreased in SD and SD+ Srt groups ( P<0.05). Compared with group SD, the escape latency was significantly shortened, the time of staying at the platform quadrant was prolonged, and the frequency of crossing the platform was increased on 3 and 4 days, the apoptotic rate and neuronal degeneration rate were decreased, the expression of SIRT1 and Nrf2 was up-regulated, ROC content was decreased, and SOD content was increased in group SD+ Srt ( P<0.05). Conclusions:Sleep deprivation can induce oxidative stress response in hippocampus by inhibiting the activation of SIRT1/Nrf2 signaling pathway, leading to cognitive dysfunction in rats.