Objective:To establish ICP-MS method for determining the Class 1 and Class 2A elemental impurities in pharmaceutical excipients polysorbates according to ICH Q3D(R2)Guidelines.Class 1 elemental impurities include As,Cd,Hg and Pb;Class 2A elemental impurities include Co,Ni and V.Methods:The analysis was performed using ICP-MS with the following parameters.Plasma mode:HMI-8,carrier gas:He,sampling depth:10.0 mm.RF power:1 600 W,the carrier gas flow,the nebulizer gas flow and the auxiliary gas flow were all 0.8 L·min-1.The plasma gas flow was 15.0 L·min-1.The speed of nebulizer pump was0.10 r·min-1.The temperature of atomizer chamber was 2.0℃.Analytical method:internal standard method.Results:The correla-tion coefficient for all elemental impurities was not less than 0.999.Spiked recoveries ranged between 85%-115.0%,with RSD is not more than 5.0%.The RSD of precision rate was not more than 10.0%.The contents of 7 elemental impurities in 15 batches of the samples were all less than PDE for injection set by ICH Q3D(R2)guidelines.Conclusion:The developed ICP-MS method demonstrates high sensitivity,good precision,and accuracy,making it suitable for controlling the Class 1 and Class 2A elemental impurities in polysorbate pharma-ceutical excipients.The results shows low risks of elemental impurities in commercial pharmaceutical excipients polysorbates.

Objective:To establish ICP-MS method for determining the Class 1 and Class 2A elemental impurities in pharmaceutical excipients polysorbates according to ICH Q3D(R2)Guidelines.Class 1 elemental impurities include As,Cd,Hg and Pb;Class 2A elemental impurities include Co,Ni and V.Methods:The analysis was performed using ICP-MS with the following parameters.Plasma mode:HMI-8,carrier gas:He,sampling depth:10.0 mm.RF power:1 600 W,the carrier gas flow,the nebulizer gas flow and the auxiliary gas flow were all 0.8 L·min-1.The plasma gas flow was 15.0 L·min-1.The speed of nebulizer pump was0.10 r·min-1.The temperature of atomizer chamber was 2.0℃.Analytical method:internal standard method.Results:The correla-tion coefficient for all elemental impurities was not less than 0.999.Spiked recoveries ranged between 85%-115.0%,with RSD is not more than 5.0%.The RSD of precision rate was not more than 10.0%.The contents of 7 elemental impurities in 15 batches of the samples were all less than PDE for injection set by ICH Q3D(R2)guidelines.Conclusion:The developed ICP-MS method demonstrates high sensitivity,good precision,and accuracy,making it suitable for controlling the Class 1 and Class 2A elemental impurities in polysorbate pharma-ceutical excipients.The results shows low risks of elemental impurities in commercial pharmaceutical excipients polysorbates.

Objective:To explore the mechanism of ubiquitin-specific protease 11(USP11)affecting the proliferation and invasion of oral squamous cell carcinoma(OSCC)cells by regulating IGF2BP3 expression.Methods:USP11 expression in OSCC tissues and adja-cent tissues from OSCC patients(n=50)was detected by immunohistochemistry and western blot,and USP11 expression in normal hu-man oral keratinocyte(HOK)cell line and human OSCC cell lines SCC-25 and CAL-27 was detected by western blot.SCC-25 and CAL-27 cells were transfected with siRNA USP11(si-USP11)or siRNA negative control(si-NC).Western blot was performed to de-tect the silencing efficiency of USP11.CCK-8,wound healing assay and Transwell assay were carried out to evaluate the effects of USP11 silencing on cell proliferation,migration and invasion.Western blot was employed to detect IGF2BP3 expression after the knockdown of USP11.Nude mice were inoculated with SCC-25 cells to construct the transplanted tumor model,and the inhibitory effect of USP11 knock-down on SCC-25 cell tumorigenicity was investigated.Results:The USP11 protein level in carcinoma tissues of OSCC patients was significantly higher than in the adjacent tissues,USP11 protein expression was significantly higher in SCC-25 and CAL-27 cells than in HOK cells.The knockdown of USP11 markedly reduced the proliferation,migration and invasion of SCC-25 and CAL-27 cells,and down-regulated the expression of IGF2BP3 cells.Compared with the USP11 silencing group,the proliferation,migration and invasion of SCC-25 and CAL-27 cells were significantly increased in the simultaneous knockdown of USP11 and overexpression of IGF2BP3 cells.Compared with the USP11 overexpression group,the proliferation,migration and invasion of SCC-25 and CAL-27 cells were decreased in the simultaneous IGF2BP3 knockdown and USP11 overexpression cells.Tumorigenicity experiments in nude mice showed that the tumor volume and weight were significantly declined by USP11 knockdown.Conclusion:USP11 is highly expressed in OSCC tissues,which may promote the proliferation,migration and invasion of OSCC cells through up-regulation of IGF2BP3 expression.

Background and purpose:Thyroid dysfunction can frequently be discovered in breast cancer patients during long-term follow-up after receiving post-operative radiation therapy(PORT).This study aimed to compare serum levels of thyroid transcription factors(TTFs)TTF-1 and paired box 8(PAX-8)before and after PORT in breast cancer patients,combined with the results of serological thyroid indicators tests,and to analyze the relationship between the changes in serum levels of these two kinds of TTFs and thyroid dysfunction after breast cancer PORT.Methods:Female breast cancer patients without thyroid disease records who received PORT in Department of Radiation Oncology,Shanghai Ninth People's Hospital Huangpu Branch,Shanghai Jiao Tong University School of Medicine from Jan.2022 to Jun.2022 were prospectively selected,and were divided into two groups according to being with or without supraclavicular radiation field.All the patients had given informed consent before joining the study.The study design was approved by the ethic committee of our hospital(Ethic Approval No.2021-KY-2).Serum levels of TTF-1 and PAX-8,serological thyroid indicators[triiodothyronine(T3),tetraiodothyronine(T4),free triiodothyronine(FT3),free tetraiodothyronine(FT4),thyroid stimulating hormone(TSH)and thyroid peroxidase antibody(TPO-Ab)]were recorded before PORT,at the end of PORT,6,12 and 24 months after the end of PORT,respectively.The Strengthening the Reporting of Observational Studies in Epidemiology(STROBE)checklist was followed for this study.Results:Eighty patients were enrolled in this study(40 in each group).A total of 19 patients who had hypothyroidism were divided in two groups,15 in supraclavicular field group(SC group)and 4 in non-supraclavicular field group(NSC group),respectively(P=0.004).Levels of TTF-1[5.70(5.11,7.13)vs 6.68(5.15,7.57),P=0.296]and PAX-8(5.26±1.01 vs 5.66±1.37,P=0.149)did not show statistically significant deference between two groups before PORT.In SC group,levels of TTF-1 and PAX-8 gradually rose in 12 months after the end of PORT.In NCS group,levels of TTF-1 and PAX-8 did not change significantly during 24 months after the end of PORT.Test results of serum TTF-1 between two groups were statistically different at 6 months[6.99(4.73,13.94)vs 5.79(5.01,6.28),P=0.049],12 months[7.65(5.02,17.85)vs 5.43(4.52,6.22),P=0.005]after the end of PORT,while test results of serum PAX-8 between two groups were statistically different at 12 months[6.79(4.86,14.30)vs 5.81(4.70,7.25),P=0.042]after the end of PORT.The median values of TTF-1 and PAX-8 test results at 12 months after the end of PORT in SC group which were both significantly higher compared with NSC group were selected as the referent thresholds.Patients in SC group whose test results were higher than referent thresholds were defined as TTF-1/PAX-8 elevating subgroups,and patients whose test results under the threshold defined as TTF-1/PAX-8 normal subgroups.The incidences of hypothyroidism were higher in elevation subgroups than in normal subgroups(65.0%vs 10.0%,60.0%vs 15.0%,respectively,P=0.001,P=0.008,respectively).Positive correlations were observed between the elevation of TTF-1/PAX-8 at 12 months after the end of PORT and hypothyroidism after breast cancer supraclavicular field radiation(OR=9.702,3.930,and P=0.020,0.046,respectively)according to multivariate analysis.Conclusion:Thyroid dysfunction after breast cancer PORT was mainly manifested with hypothyroidism;supraclavicular field radiation may significantly increase the incidence of hypothyroidism;serum levels of TTF-1 and PAX-8 elevated obviously in breast cancer PORT patients who had hypothyroidism after receiving supraclavicular field radiation.

Background and purpose:Thyroid dysfunction can frequently be discovered in breast cancer patients during long-term follow-up after receiving post-operative radiation therapy(PORT).This study aimed to compare serum levels of thyroid transcription factors(TTFs)TTF-1 and paired box 8(PAX-8)before and after PORT in breast cancer patients,combined with the results of serological thyroid indicators tests,and to analyze the relationship between the changes in serum levels of these two kinds of TTFs and thyroid dysfunction after breast cancer PORT.Methods:Female breast cancer patients without thyroid disease records who received PORT in Department of Radiation Oncology,Shanghai Ninth People's Hospital Huangpu Branch,Shanghai Jiao Tong University School of Medicine from Jan.2022 to Jun.2022 were prospectively selected,and were divided into two groups according to being with or without supraclavicular radiation field.All the patients had given informed consent before joining the study.The study design was approved by the ethic committee of our hospital(Ethic Approval No.2021-KY-2).Serum levels of TTF-1 and PAX-8,serological thyroid indicators[triiodothyronine(T3),tetraiodothyronine(T4),free triiodothyronine(FT3),free tetraiodothyronine(FT4),thyroid stimulating hormone(TSH)and thyroid peroxidase antibody(TPO-Ab)]were recorded before PORT,at the end of PORT,6,12 and 24 months after the end of PORT,respectively.The Strengthening the Reporting of Observational Studies in Epidemiology(STROBE)checklist was followed for this study.Results:Eighty patients were enrolled in this study(40 in each group).A total of 19 patients who had hypothyroidism were divided in two groups,15 in supraclavicular field group(SC group)and 4 in non-supraclavicular field group(NSC group),respectively(P=0.004).Levels of TTF-1[5.70(5.11,7.13)vs 6.68(5.15,7.57),P=0.296]and PAX-8(5.26±1.01 vs 5.66±1.37,P=0.149)did not show statistically significant deference between two groups before PORT.In SC group,levels of TTF-1 and PAX-8 gradually rose in 12 months after the end of PORT.In NCS group,levels of TTF-1 and PAX-8 did not change significantly during 24 months after the end of PORT.Test results of serum TTF-1 between two groups were statistically different at 6 months[6.99(4.73,13.94)vs 5.79(5.01,6.28),P=0.049],12 months[7.65(5.02,17.85)vs 5.43(4.52,6.22),P=0.005]after the end of PORT,while test results of serum PAX-8 between two groups were statistically different at 12 months[6.79(4.86,14.30)vs 5.81(4.70,7.25),P=0.042]after the end of PORT.The median values of TTF-1 and PAX-8 test results at 12 months after the end of PORT in SC group which were both significantly higher compared with NSC group were selected as the referent thresholds.Patients in SC group whose test results were higher than referent thresholds were defined as TTF-1/PAX-8 elevating subgroups,and patients whose test results under the threshold defined as TTF-1/PAX-8 normal subgroups.The incidences of hypothyroidism were higher in elevation subgroups than in normal subgroups(65.0%vs 10.0%,60.0%vs 15.0%,respectively,P=0.001,P=0.008,respectively).Positive correlations were observed between the elevation of TTF-1/PAX-8 at 12 months after the end of PORT and hypothyroidism after breast cancer supraclavicular field radiation(OR=9.702,3.930,and P=0.020,0.046,respectively)according to multivariate analysis.Conclusion:Thyroid dysfunction after breast cancer PORT was mainly manifested with hypothyroidism;supraclavicular field radiation may significantly increase the incidence of hypothyroidism;serum levels of TTF-1 and PAX-8 elevated obviously in breast cancer PORT patients who had hypothyroidism after receiving supraclavicular field radiation.

Objective:To analyze the relationship between PD-L1 expression and the degree of infiltration of different types of TILs in triple-negative breast cancer, and the correlation with other pathological features to explore their clinical significance.Methods:Tumor tissues of 199 patients with triple-negative breast cancer in our hospital were selected, and the expression of PD-L1 and the distribution of TILs were analyzed using immunohistochemistry and counted using Image software. The relationship between CD8 +TILs, CD4 +TILs and PD-L1 expression was analyzed and compared, and survival analysis and correlation analysis between PD-L1 expression and pathological information were performed. Results:The density of CD4 + TILs ( χ2=8.75, P=0.003) and CD8 + TILs ( χ2=6.32, P=0.009) infiltration was higher in the PD-L1-positive group compared to that of the PD-L1-negative group. Among PD-L1-positive patients, patients with higher TILs infiltration compared to low infiltrating TILs could achieve longer overall survival time, and the difference was statistically significant ( P1=0.012, P2=0.023, P3=0.010). Patient PD-L1 expression was positively correlated with the number of positive lymph nodes, tumor stage, and ki-67 expression ( Pa=0.032, Pb=0.006, Pc=0.042), and was not related to age or tumor diameter ( Pd=1.031, Pe=0.672) . Conclusions:PD-L1 expression in triple negative breast cancer predicts higher infiltration of CD4 +TILs and CD8 +TILs, while higher infiltration of CD4 +TILs and CD8 +TILs predicts a relatively good survival prognosis.PD-L1 expression is associated with multiple pathological clinical factors and deserves further study to make PD-L1, TILs and other indicators better benefit triple-negative breast cancer patients.

GI (GIGANTEA) is one of the output key genes for circadian clock in the plant. The JrGI gene was cloned and its expression in different tissues was analyzed to facilitate the functional research of JrGI. RT-PCR (reverse transcription-polymerase chain reaction) was used to clone JrGI gene in present study. This gene was then analyzed by bioinformatics, subcellular localization and gene expression. The coding sequence (CDS) full length of JrGI gene was 3 516 bp, encoding 1 171 amino acids with a molecular mass of 128.60 kDa and a theoretical isoelectric point of 6.13. It was a hydrophilic protein. Phylogenetic analysis showed that JrGI of 'Xinxin 2' was highly homologous to GI of Populus euphratica. The result of subcellular localization showed that JrGI protein was located in nucleus. The JrGI, JrCO and JrFT genes in female flower buds undifferentiated and early differentiated of 'Xinxin 2' were analyzed by RT-qPCR (real-time quantitative PCR). The results showed that the expression of JrGI, JrCO and JrFT genes were the highest on morphological differentiation, implying the temporal and special regulation of JrGI in the differential process of female flower buds of'Xinxin 2'. In addition, RT-qPCR analysis showed that JrGI gene was expressed in all tissues examined, whereas the expression level in leaves was the highest. It is suggested that JrGI gene plays a key role in the development of walnut leaves.
Juglans/genetics* ; Phylogeny ; Plant Leaves ; Cloning, Molecular ; Gene Expression Regulation, Plant ; Plant Proteins/metabolism*

Objective:To compare the clinical effects of percutaneous curved vertebroplasty (PCVP) and unilateral percutaneous kyphoplasty (PKP) in the treatment of osteoporotic vertebral compression fracture (OVCF).Methods:A retrospective cohort study was used to analyze the clinical data of 104 patients with single vertebral OVCF treated in Tianjin Hospital from September 2019 to September 2020, including 21 males and 83 females; aged 50-91 years [(70.3±7.7)years]. AO classification of the fracture was type A1 in 65 patients and type A2 in 39. The patients received PCVP (PCVP group, n=51) or unilateral PKP surgery (unilateral PKP group, n=53). The operation time, bone cement injection volume, intraoperative fluoroscopy frequency, effective dispersion times of bone cement and excellent rate of bone cement distribution were compared between the two groups. In evaluation of the therapeutic effects of the two groups, visual analogue scale (VAS) and Oswestry dysfunction index (ODI) were measured preoperatively and at postoperative 24 hours, 3 months and 6 months; Beck index was measured preoperatively and at postoperative 24 hours and 3 months. The rate of bone cement leakage and rate of refracture of adjacent vertebral bodies were compared between the two groups. Results:All patients were followed up for 6-8 months [(6.4±0.7)months]. The operation time, bone cement injection volume and intraoperative fluoroscopy frequency in PCVP group was (12.15±1.63)minutes, (2.13±0.28)ml and (24.74±1.71)times, shorter or less than (22.09±1.62)minutes, (5.30±0.52)ml and (30.09±1.86)times in unilateral PKP group (all P<0.01). The effective dispersion times of bone cement in PCVP group was (1.42±0.04)times, higher than (1.18±0.02)times in unilateral PKP group ( P<0.01). The excellent rate of bone cement distribution in PCVP group was 94%, higher than 70% in unilateral PKP group ( P<0.01). There were no significant differences in VAS, ODI and Beck index between the two groups before operation and at 24 hours and 3 months after operation (all P>0.05). VAS and ODI in PCVP group were (1.20±0.49)points and 16.52±5.22 at 6 months after operation, lower than (1.49±0.58)points and 20.16±5.16 in unilateral PKP group (all P<0.01). VAS and ODI in the two groups were significantly improved at 24 hours, 3 months and 6 months after operation when compared with those before operation (all P<0.05). Beck index in the two groups detected at 24 hours and 3 months after operation was improved from that before operation (all P<0.05). Unilateral PKP group showed Beck index was 0.75±0.07 at 3 months after operation, significantly lower than 0.79±0.07 at 24 hours after operation ( P<0.05), but there was no significant change in PCVP group ( P>0.05). The leakage rate of bone cement in PCVP group was 16% (8/51), lower than 47% (25/53) in unilateral PKP group ( P<0.01). There was no significant difference in the incidence of refracture of adjacent vertebral bodies between the two groups during follow-up ( P>0.05). Conclusion:For OVCF, PCVP is superior to unilateral PKP in terms of operation time, amount of bone cement injection, intraoperative fluoroscopy frequency, dispersion effect of bone cement in vertebral body, pain, function improvement, maintenance of injured vertebral height and incidence of bone cement leakage.

【Objective】 To study the changes of blood coagulation function of donors before and after peripheral blood stem cell(PBSC)mobilization and collection, so as to evaluate the safety of the current scheme. 【Methods】 30 donors who received PBSC mobilization and collection in Zhujiang Hospital from October 2018 to October 2020 were enrolled. After mobilization by G-CSF, the correlation between coagulation function, blood routine indexes and TEG indexes of donors was analyzed, and the influence of PBSC mobilization and collection on coagulation function of donors was evaluated. 【Results】 The TEG indexes R(min), K(min), α(°), MA(mm) and CI before and after PBSC collection were 6.12±1.18 vs 7.25±2.16, 1.98±0.41 vs 2.45±0.64, 62.82±4.98 vs 57.3±6.67, 60.93±3.26 vs 55.37±4.41, and -0.31±1.40 vs -2.32±2.18, respectively(P<0.05), suggesting that there was no risk of hypercoagulability after PBSC mobilization and collection. The peak values of WBC (×10⁹/L), Plt (×10⁹/L) and Hb (g/L) were 62.02, 357 and 162, respectively, which indicated that the blood routine indexes after PBSC mobilization and collection were in the safe range. After PBSC collection, the CI value of 26.7% (8/30) donors was less than -3, showing hypocoagulability. 【Conclusion】 The current mobilization and collection scheme of PBSC has little effect on the coagulation function. Most of the donors had no risk of hypercoagulability, but a few showed a trend of hypocoagulability after PBSC collection.

【Objective】 To explore the effects of blood routine parameters on the peripheral blood hematopoietic stem cell collection of healthy donors, and predict collection timing based on these parameters. 【Methods】 The blood routine parameters pre-donation and the total number of mononuclear cells post-donation of 249 donors who applied blood cell separator to collect peripheral blood hematopoietic stem cells in our hospital from January 2018 to August 2020 were collected. Taking total nucleated cells of circulating blood per litre as the main evaluation index, and its collection with blood routine parameters pre-collection was analyzed. The relevant influencing factors were analyzed using multiple linear regression analysis. The blood routine parameters of healthy donors who donated peripheral blood hematopoietic stem cells in our hospital from September 2020 to October 2020 were substituted into the equation to obtain the predicted values, which were then compared with the actual values obtained from actual product using t test for verification. 【Results】 The analysis showed that the parameters of Hb, RBC, Hct, leukocyte count, neutrophil, lymphocyte, monocyte and Plt were statistically correlated with the total number of mononuclear cells of circulating blood per liter volume (P<0.05). There was a linear relationship between lymphocyte, monocyte, Plt and leukocyte count and the total number of mononuclear cells of circulating blood per liter. The total number of mononuclear cells of circulating blood per liter was set to (Y), and the variables such as lymphocyte (X1), monocyte (X2), Plt (X3), leukocyte count (X), and neutrophil were used as dependent variables for multiple linear regression, and the equation was: Y=9.814+ 3.131X1+ 1.666X2+ 0.020X3+ 0.124X4. There was no statistical difference between the predicted value and the calculated value (P>0.05). 【Conclusion】 The blood routine parameters of lymphocyte, monocyte, platelet count and leukocyte count of donors before collection can effectively predict the collection efficiency, therefore help predict the collection time.