Objective: To explore the setting of gray zone of Treponema pallidum (TP) testing by retrospective analysis of blood donors with single reagent reactive anti-TP results, so as to improve blood utilization and supply safety. Methods: Blood samples were collected from 112 blood donors previously deferred due to single reagent reactive TP antibody results between January 2020 and December 2023, and subjected to dual ELISA reagents and TPPA test. The gray zone panel analysis was performed on the two ELISA reagents currently used in our department. The detection rate at each concentration of the gray zone panle was counted, and the corresponding concentrations for C , C , and C and gray zone cut-off were calculated. Results: Among the 50 samples deferred by reagent 1, 19 were confirmed reactive and 31 non-reactive in supplementary testing. Among the 62 samples deferred by reagent 2, 12 were confirmed reactive and 50 non-reactive in supplementary testing. For reagent 1, the detection rate of was 56% for S/CO≥1 and 20% for 0.5≤S/CO<1, retrospectively. For reagent 2, the detection rate was 27% for S/CO≥1 and 12.5% for 0.5≤S/CO<1, retrospectively. The detection rate for S/CO≥1 was higher than those for 0.5≤S/CO<1 for both reagents. All the 112 samples were negative in TPPA test. The C concentration of reagent 1 was 1.51 mIU/mL, and the concentration range of C ±20% was 1.21-1.81 mIU/mL. The C concentration of reagent 2 was 1.45 mIU/mL, and the concentration range of C ±20% was 1.16-1.74 mIU/mL. The C and C concentration of both reagents were within the C ±20% range, suggesting that the gray zone cutoff for both Reagent 1 and Reagent 2 should be set at S/CO=0.8 (80% of the CO value). Conclusion: All anti-TP single reagent reactive samples with S/CO value within the gray zone was tested negative by TPPA. It is necessary to consider the rationality and necessity of establishing the gray zone, so as to ensure blood safety and improve the utilization rate of blood resources.

OBJECTIVE To investigate the marketing status， general characteristics， and time trends of rare disease drugs in China. METHODS Based on 121 kinds of rare diseases included in the First Batch of Rare Disease Catalog， the names and marketing approval information of corresponding drugs with indications were obtained from the databases of the Center for Drug Evaluation， National Medical Products Administration and Yaozhi.com， and the relevant characteristic variables were extracted for descriptive statistical analysis. RESULTS As of December 31， 2022， only 32 of 121 rare diseases have therapeutic drugs available for treatment on the market in China， and 79 rare disease drugs have been approved. Among them， 46.84% of the drugs are domestic drugs， 88.61% of the drugs are approved for use in both adults and children； 67.09% are chemicals and 59.49% are injections. According to the ATC classification， Category A （digestive system drugs） is the most， accounting for 20.25%. The number of rare disease drugs on the market each year is the highest in 2021， with an overall upward trend from 2018 to 2021 and a downward trend in 2022. Among rare disease drugs on the market each year， according to the ATC classification， the number of Category L （antineoplastics and immune inhibitors） will be the largest in 2021， being 5. By dosage form， oral medicines were marketed in the largest number in 2022， and injectable medicines in 2021. CONCLUSIONS In recent years， the number of approved rare disease drugs in China has been continuously increasing， but it is still far from meeting the needs of patients， and there is still a lack of domestically approved rare disease drugs. We should further accelerate the research and development of rare disease drugs， and promote the import and replication of rare disease drugs.

OBJECTIVE To investigate the marketing status， general characteristics， and time trends of rare disease drugs in China. METHODS Based on 121 kinds of rare diseases included in the First Batch of Rare Disease Catalog， the names and marketing approval information of corresponding drugs with indications were obtained from the databases of the Center for Drug Evaluation， National Medical Products Administration and Yaozhi.com， and the relevant characteristic variables were extracted for descriptive statistical analysis. RESULTS As of December 31， 2022， only 32 of 121 rare diseases have therapeutic drugs available for treatment on the market in China， and 79 rare disease drugs have been approved. Among them， 46.84% of the drugs are domestic drugs， 88.61% of the drugs are approved for use in both adults and children； 67.09% are chemicals and 59.49% are injections. According to the ATC classification， Category A （digestive system drugs） is the most， accounting for 20.25%. The number of rare disease drugs on the market each year is the highest in 2021， with an overall upward trend from 2018 to 2021 and a downward trend in 2022. Among rare disease drugs on the market each year， according to the ATC classification， the number of Category L （antineoplastics and immune inhibitors） will be the largest in 2021， being 5. By dosage form， oral medicines were marketed in the largest number in 2022， and injectable medicines in 2021. CONCLUSIONS In recent years， the number of approved rare disease drugs in China has been continuously increasing， but it is still far from meeting the needs of patients， and there is still a lack of domestically approved rare disease drugs. We should further accelerate the research and development of rare disease drugs， and promote the import and replication of rare disease drugs.

Objective To investigate the expression of PIK3CA,phosphorylated protein kinase B(p-AKT)and phosphatase and tensin homologue deleted on chromosome 10(PTEN)in sinonasal squamous cell carcinoma(SNSCC).Methods The expressions of PIK3CA and PTEN in head and neck squamous cell carci-noma(HNSCC)were analyzed through the data set of HNSCC in the cancer genome map of UCSC Xena data-base.The immunohistochemical SP method was used to measure the expression of PIK3CA,p-AKT and PTEN in 43 cases of SNSCC tissues,20 cases of normal inferior concha tissues.The relationship between the expressions of PIK3CA,p-AKT and PTEN protein with the clinicopathological features and prognosis of the patients with SNSCC was analyzed.Results The results of bioinformatic analysis showed that PIK3CA mR-NA expression in HNSCC tissues was higher than that in paracancerous tissues(P＜0.01),while the PTEN mRNA expression was lower than that in paracancerous tissues(P＜0.05).The immunohistochemical detec-tion results showed that the positive expressions rates of PIK3CA and p-AKT proteins in normal nasal mucosa tissues were significantly lower than those in SNSCC tissues,while the positive expression rate of PTEN pro-tein in SNSCC tissues was significantly higher than that in normal inferior nasal concha mucosa tissues,and the differences were statistically significant(P＜0.01).The expressions of PIK3CA and p-AKT protein were related to the clinical stage,differentiation degree and primary site(P＜0.05),but were not related to age,gender,smoking and drinking(P＞0.05);the PTEN protein expression was not related with the clinical stage,differentiation degree,primary site,age,smoking and drinking(P＞0.05).The Spearman analysis showed that the expression of PIK3CA in SNSCC tissues was positively correlated with p-AKT protein ex-pression(r=0.664,P＜0.01),and PIK3CA was negatively correlated with PTEN protein(r=-0.414,P＜0.01).The expression of p-AKT was negatively correlated with PTEN protein(r=-0.453,P＜0.01).The Kaplan-Meier analysis showed that the median survival time of the patients with PIK3CA and p-AKT protein positive expression was shorter than that of the patients with negative expression(P＜0.01).There was no statistically significant difference in median survival between the patients with PTEN protein positive expres-sion and those with negative expression.Conclusion The overexpressions of PIK3CA and p-AKT accompa-nied by the loss of PTEN expression participate in the development and progression of SNSCC,moreover the PIK3CA and p-AKT expressions are related to the poor prognosis of the patients.

BACKGROUND:Growth differentiation factor 5 is a member of the transforming growth factor superfamily and one of the earliest markers of joint development.Growth differentiation factor 5 has an important role in cartilage repair. OBJECTIVE:To explore the action mechanism of growth differentiation factor 5-induced chondrogenic differentiation of bone marrow mesenchymal stem cells. METHODS:Rabbit bone marrow mesenchymal stem cells were isolated and cultured.CCK-8 assay was used to detect the effect of different mass concentrations of growth differentiation factor 5 on the proliferation activity of bone marrow mesenchymal stem cells.RT-PCR was utilized to detect the expression of genes related to chondrogenic differentiation of bone marrow mesenchymal stem cells induced by different mass concentrations of growth differentiation factor 5.To further investigate the action mechanism of growth differentiation factor 5-induced chondrogenic differentiation of bone marrow mesenchymal stem cells,we added inhibitor XAV-939 and activator Laduviglusib of Wnt/β-catenin signaling pathways to induce cell culture for 14 days.RT-PCR and western blot assay were performed to detect the expression of cartilage-related genes and Wnt/β-catenin signaling pathway proteins. RESULTS AND CONCLUSION:(1)CCK-8 results showed no significant effect of growth differentiation factor 5 on the proliferation of bone marrow mesenchymal stem cells.(2)Growth differentiation factor 5 promoted the expression of cartilage-related genes type Ⅱ collagen,aggrecan and Sox9,among which growth differentiation factor 5 induced a significant upregulation of cartilage-related genes in the 50 ng/mL group.(3)Addition of Laduviglusib,an activator of Wnt/β-catenin signaling pathway,upregulated Sox9,β-catenin and type Ⅱ collagen expression(P<0.05).Addition of XAV939,an inhibitor of Wnt/β-catenin signaling pathway,down-regulated Sox9,β-catenin and type Ⅱ collagen expression(P<0.05).(4)Taken together,growth differentiation factor 5-induced chondrogenic differentiation of bone marrow mesenchymal stem cells may be associated with the activation of the Wnt/β-catenin signaling pathway.

BACKGROUND:Growth differentiation factor 5,a member of the transforming growth factor-β superfamily and bone morphogenetic protein family,plays an important role in articular cartilage injury repair,bone regeneration,improvement of intervertebral disc degeneration,tendon healing,and neurodevelopment. OBJECTIVE:To review the research progress of growth differentiation factor 5 in inducing chondrocytes,nucleus pulposus-like cells,tendon cell differentiation,as well as inducing bone formation and neurodevelopment. METHODS:The search terms"growth differentiation factor 5,articular cartilage,bone,nucleus pulposus cells,tendon,nerve regeneration"were used in CNKI,WanFang and PubMed.According to the inclusion and exclusion criteria,the articles not related to the subject matter were excluded and 69 articles related to growth differentiation factor 5 were included. RESULTS AND CONCLUSION:(1)Growth differentiation factor 5 can induce chondrogenic and osteoblastic differentiation of mesenchymal stem cells,but the concentration boundary of growth differentiation factor 5 to induce chondrogenic or osteoblastic differentiation remains unclear.(2)Growth differentiation factor 5 can induce mesenchymal stem cells to differentiate into nucleus pulposus cells,which may play a role in the treatment of intervertebral disc degeneration.(3)Growth differentiation factor 5 can induce mesenchymal stem cells to differentiate into tendon cells and play an important role in tendon repair and prevention of postoperative tendon adhesion.(4)Growth differentiation factor 5 can induce neurodevelopment and promote nerve regeneration.

Purpose To explore the feasibility of artificial intelligence compressed sensing(ACS)technique in ankle joint MRI.Materials and Methods From September to October 2023,32 healthy volunteers who underwent ankle joint scanning in Beijing Friendship Hospital,Capital Medical University were prospectively collected.MRI of the ankle joint based on ACS and parallel imaging(PI)technology was performed on 3.0T MR.The sagittal proton density weighted imaging(PDWI),coronary PDWI,transverse PDWI and sagittal T1WI were acquired,and all data were divided into test group and control group,with ACS to accelerate the multiples of 5(ACS 5.0)in test group,whereas PI speed ratio of 2(PI 2.0)in control group,respectively.The signal intensity of talus,achilles tendon and cartilage were measured,the signal intensity and standard deviation of the long hallux flexor were obtained,and the signal noise ratio(SNR)and contrast to noise ratio(CNR)were calculated via long hallux flexor as background noise.The data of objective and subjective evaluation of the two sequences were statistically analyzed,and the image quality of each sequence was evaluated via the standard reference of PI 2.0.Results SNR and CNR in ACS group were higher than those in PI group,and the anatomical structure of sagittal PDWI sequence between the two groups had statistical significance(t=-2.937,-1.981,-4.058,-3.879,P<0.05).There were significant differences in cartilage SNR and talus CNR in coronal PDWI sequence(t=-3.310,-3.567;P=0.002,P<0.001).In terms of axial PDWI sequence,there were statistically significant differences in talus CNR and cartilage CNR between ACS and PI groups(t=-4.270,-4.382,P<0.05).The subjective evaluation of the image quality scores of the two groups by the two diagnostic imaging doctors showed a strong observer consistency(Kappa=0.977,P=0.009).There was no significant difference in image quality scores between the two groups(Z=-0.248,-0.747,<0.001,-0.071,P>0.05).The total collection time of ACS group and PI group was 337 s and 610 s,respectively.Compared with PI group,the total scanning time of ACS group was shortened by 44.8%.Conclusion ACS based MRI of the ankle joint can not only shorten the scan time,but also ensure and further improve the image quality,with feasibility.

Objective To investigate the relationship between nestin expression and microvessel density(MVD)and its value as a potential prognostic marker of glioblastoma(GBM).Methods A retrospective trial was conducted on 50 GBM patients undergoing craniotomy under general an-esthesia in the Third Affiliated Hospital of Chongqing Medical University from March 2020 to March 2023.According to the median nestin H score(Nestin staining intensity × percentage of positive cells),they were divided into H score ≥80.0 group(28 cases)and H score＜80.0 group(22 cases).After 2 years of follow-up,they were also divided into a death group(23 cases)and a survival group(27 cases).The expression of nestin was determined by immunohistochemical staining,and MVD was quantified by von Willebrand factor(vWF)and CD105 immunohistochem-ical staining.Spearman rank test was used to analyze the correlation between nestin expression level and MVD formation,multivariate Cox proportional hazards regression analysis was em-ployed to identify the influencing factors for overall survival in the GBM patients,and Kaplan-Meier survival curve was plotted to analyze the 2-year survival rate of the patients.Results The nestin H score was positively correlated with vWF-MVD(Rho=0.701,P＜0.01)and CD105-MVD(Rho=0.753,P＜0.01).There were no significant differences between the H score ≥80.0 group and the H score＜80.0 group in terms of gender,age,isocitric dehydrogenase-1 mutation,tumor site,tumor location,brain necrosis,tumor volume,edema volume,preoperative Karnofsky Performance Status score,and extent of tumor resection(P＞0.05).The survival group had larger proportions of isocitrate dehydrogenase-1 mutation and maximal resection,and lower nestin H score than the death group(P＜0.05,P＜0.01).Cox proportional hazards regression showed that partial resection/biopsy(HR=4.781,95％CI:2.066-11.060,P=0.001)and increased nestin H score(HR=1.007,95％CI:1.001-1.013,P=0.032)were independent influencing factors for worsening overall survival in the GBM patients.Kaplan-Meier survival curve analysis indicated that the cumulative survival rate was lower in the H score ≥80.0 group than the H score＜80.0 group(log rank x2=8.147,P=0.004).Conclusion Nestin overexpression is associated with poor prognosis in GBM patients,indicating more microangiogenesis,which may be a therapeutic target for GBM patients.

Objective:To explore the efficacy and safety of cryopreservation-free integrated autologous hematopoietic stem cell transplantation (HSCT) model for patients with multiple myeloma.Methods:A total of 96 patients with newly diagnosed multiple myeloma (NDMM) between July 31, 2020, and December 31, 2022, were retrospectively analyzed, of which 41 patients in the observation group received integrated non-cryopreserved transplantation mode. After hematopoietic stem cells were mobilized and collected, melphalan was started immediately for pre-transplant conditioning, and non-cryopreserved grafts from the medical blood transfusion refrigerator were directly injected intravenously into the patient within 24-48 h after the melphalan conditioning. The control group consisted of 55 patients who received traditional transplantation mode. After hematopoietic stem cells were collected, stem cell cryopreservation was performed in liquid nitrogen, and then the transplant plans were started at the right time. All patients received mobilization of autologous hematopoietic stem cells using the G-CSF combined with the plerixafor.Results:① A total of 34 patients (82.9% ) with VGPR plus CR in the observation group were significantly higher than 33 patients (60.0% ) in the control group ( P=0.016). ②Compared with the control group, the incidence of grade 1 oral mucosal inflammation was higher in the observation group ( P<0.001) ; however, the incidence of grades 2 and 3 oral mucosal inflammation was lower ( P=0.004, P=0.048), and neither group experienced grade 4 or above oral mucosal inflammation. The incidence of grade 1 diarrhea was higher in the observation group ( P=0.002), whereas the incidence of grade 3 diarrhea was lower ( P=0.007). No statistically significant difference was observed in the incidence of grade 4 diarrhea ( P=0.506), and neither group experienced grade 5 diarrhea. ③ The incidence of bacterial infection in the observation group was lower than that in the control group (34.1% vs 65.5%, P=0.002), whereas no statistically significant difference was observed in the incidence of fungal infection (29.3% vs 31.4%, P=0.863) and viral infection (4.88% vs 3.64%, P=0.831). ④No statistically significant difference was observed in the implantation time of granulocytes and platelets between the observation and control groups [10 (8-20) days vs 11 (8-17) days, P=0.501; 13 (10-21) days vs 15 (10-20) days, P=0.245]. ⑤ All patients did not receive lenalidomide treatment 100 days post-transplantation. At 30 days post-transplantation, the CTL, NK, and Th cell counts in the observation group were lower than those in the control group ( P<0.001, P=0.002, P=0.049), and the NKT cell counts were higher than those in the control group ( P=0.024). At 100 days post-transplantation, the CTL, NKT, and Th cell counts in the observation group were higher than those in the control group ( P=0.025, P=0.011, P=0.007), and no statistically significant difference in NK cell counts was observed between the two groups ( P=0.396). ⑥ The median follow-up was 18 (4-33) months. The overall 2-year survival rates of the observation and control groups post-transplantation were 91.5% and 78.2%, respectively ( P=0.337). The recurrence-free survival rates were 85.3% and 77.6%, respectively ( P=0.386), and the cumulative recurrence rates were 9.8% and 16.9%, respectively ( P=0.373) . Conclusion:In NDMM, the cryopreservation-free integrated autologous HSCT model can achieve similar therapeutic effects as traditional transplantation models, with lower rates of severe mucosal inflammation and infection compared with traditional transplantation models.

Objective To investigate the mechanism of 2,6-dimethoxy-1,4-benzoquinone(DMQ),an active ingredients in fermented wheat germ extract,for inhibiting NLRP3 inflammasome activation and alleviating septic shock in mice.Methods Cultured murine bone marrow-derived macrophages(BMDM)stimulated with lipopolysaccharide(LPS)were treated with DMQ,followed by treatment with Nigericin,ATP,and MSU for activating the canonical NLRP3 inflammasome;the non-canonical NLRP3 inflammasome was activated by intracellular transfection of LPS,and AIM2 inflammasome was activated using Poly A:T.In human monocytic THP-1 cells,the effect of Nigericin on inflammasome activation products was examined using Western blotting and ELISA.Co-immunoprecipitation was performed to explore the mechanism of DMQ-induced blocking of NLRP3 inflammasome activation.In a male C57BL/6J mouse model of LPS-induced septic shock treated with 20 and 40 mg/kg DMQ,the levels of IL-1β and TNF-α in the serum and peritoneal lavage fluid were determined using ELISA,and the survival time of the mice within 36 h was observed.Results Treatment with DMQ effectively inhibited LPS-induced activation of canonical NLRP3 inflammasome in mouse BMDM and human THP-1 cells and also inhibited non-canonical NLRP3 inflammasome activation in mouse BMDM,but produced no significant effect on AIM2 inflammasome activation.DMQ significantly blocked the binding between ASC and NLRP3.In the mouse models of septic shock,DMQ treatment significantly reduced the levels of IL-1β in the serum and peritoneal fluid and obviously prolonged survival time of the mice.Conclusion DMQ can effectively block ASC-NLRP3 interaction to inhibit NLRP3 inflammasome activation and alleviate LPS-induced septic shock in mice.