Host cell proteins(HCPs) of biological products are endogenous host proteins expressed by host cells or produced when they die during the production of biological products. Incomplete elimination of HCPs is the main factor causing allergy or other adverse reactions. The analytical methods for HCPs mainly include SDS-PAGE, Western blot(WB), two-dimensional gel electrophoresis(2DE), ELISA, liquid chromatography, mass spectrometry and so on. Electrophoresis can provide qualitative and semi-quantitative determination for proteins, but the resolution and accuracy are low. ELISA has good reproducibility and high sensitivity, and is most widely used at present. Chromatography and mass spectrometry have high separation and resolution as well as accurate quantification, but high application cost. ELISA was used to determine HCPs in recombinant biological products in the general rules <3412>, <3413> and <3414> of Chinese Pharmacopoeia(Volume Ⅲ, 2020 edition)and United States Pharmacopoeia(USP) <1132>. Analysis of HCPs components is helpful to improve the coverage of antiHCPs antibody and the sensitivity and specificity of detection methods. Traditional analytical methods can only qualitatively analyze HCPs, but cannot perform quantitative determination and characterization. Based on electrophoresis technology and mass spectrometry technology, combined with the development trend of HCPs component analysis at home and abroad, this paper summarizes the relevant HCPs analysis method to provide a reference for the verification of HCPs in the research and production process of biological products.

Objective To evaluate the corneal biomechanical changes after femtosecond laser-assisted in situ kerato-mileusis(FS-LASIK)combined with accelerated corneal cross-linking(FS-LASIK Xtra)for the correction of high myopia u-sing Corvis ST.Methods In this prospective case-control study,185 patients(185 eyes)who underwent surgical correc-tion of high myopia in the Affiliated Eye Hospital of Shandong University of Chinese Medicine from July 2020 to July 2022 were selected,including 93 patients receiving FS-LASIK Xtra in the FS-LASIK Xtra group and 92 patients receiving FS-LASIK in the FS-LASIK group.All patients had their right eyes included in the study.During the 6-month follow-up,the uncorrect-ed visual acuity,spherical equivalent(SE),mean corneal curvature,thinnest corneal thickness,corneal biomechanical pa-rameters[deformation amplitude ratio at 2 mm(DAR 2 mm),integrated radius(IR),stiffness parameter at the first appla-nation(SP-A1),stress-strain index(SSI)],and changes in corneal biomechanical parameters before and after surgery(difference between 6 months after surgery and before surgery,namely ΔDAR 2 mm,ΔIR,ΔSP-A1,and ΔSSI)were recor-ded.Independent sample t test and Mann-Whitney U test were used to compare the data between groups.Results Three months after surgery,the SE in the FS-LASIK group and the FS-LASIK Xtra group was(-0.21±0.31)D and(-0.04±0.36)D,respectively,and the difference was statistically significant(t=3.49,P=0.001).Six months after surgery,the SE in the FS-LASIK group and the FS-LASIK Xtra group was(-0.33±0.31)D and(-0.14±0.37)D,respectively,and the difference was statistically significant(t=4.00,P＜0.001).There was no significant difference in efficacy index and safety index between the FS-LASIK group and the FS-LASIK Xtra group at 6 months after operation(both P＞0.05).There were no significant differences in DAR 2 mm,IR,SP-A1,SSI and other biomechanical parameters between the two groups before surgery(all P＞0.05).At 6 months postoperatively,ΔIR in the FS-LASIK group and the FS-LASIK Xtra group was 3.03±0.78 and 2.67±0.80,respectively;ΔSP-A1 was-35.93±12.04 and-30.43±12.44,respectively;and ΔSSI was-0.09±0.10 and-0.03±0.06,respectively.The differences between the two groups were all statistically significant(all P＜0.05).Conclusion FS-LASIK Xtra for correction of high myopia improves the stability of postoperative visual acuity and SE with good safety and efficacy,but reduces the stability of comeal biomechanical parameters compared with before surgery;still,it is better than conventional FS-LASIK,and the long-term effect needs to be further assessed.

Objective:To construct a surface display system containing various lengths of the Ag43 passenger domain for an optimal bacterial surface display of foreign protein HPV16L1.Methods:(1) Ag43 gene sequences of different lengths were inserted into pET22b vector to construct four Ag43 surface display vectors (Ag43/138, Ag43/551, Ag43/552 and Ag43/700) using PCR and subcloning strategy. (2) The generation of four HPV16L1-Ag43 fusion constructs was completed by PCR and subcloning methods. (3) HPV16L1-Ag43 fusion proteins were expressed and analyzed by SDS-PAGE. (4) The surface exposure of HPV-16L1 was verified using trypsin digestion.Results:PCR analysis and sequencing results showed that Ag43 surface display vectors and HPV16L1-Ag43 fusions were constructed successfully. SDS-PAGE showed that the expression of HPV16L1-Ag43 fusion proteins could be induced with 0.2 mmol/L IPTG and the protein content was reduced after the cells were treated with trypsin, especially the content of Ag43/700-HPV16L1 that showed a drastic reduction.Conclusions:The Ag43 surface display system was successfully constructed and could be used for a successful display of HPV16L1. This study also showed that Ag43/700 comprising only the α-helix and the β-barrel of Ag43 provided an optimal surface display for HPV16L1.

Multiple plant lineages have independently evolved sex chromosomes and variable kary-otypes to maintain their sessile lifestyles through constant biological innovation.Morus notabilis,a dioecious mulberry species,has the fewest chromosomes among Morus spp.,but the genetic basis of sex determination and karyotype evolution in this species has not been identified.In this study,three high-quality genome assemblies were generated for Morus spp.[including dioecious M.notabilis(male and female)and Morus yunnanensis(female)]with genome sizes of 301-329 Mb and were grouped into six pseudochromosomes.Using a combination of genomic approaches,we found that the putative ancestral karyotype of Morus species was close to 14 protochromosomes,and that sev-eral chromosome fusion events resulted in descending dysploidy(2n=2x=12).We also charac-terized a～6.2-Mb sex-determining region on chromosome 3.Four potential male-specific genes,a partially duplicated DNA helicase gene(named MSDH)and three Ty3_Gypsy long terminal repeat retrotransposons(named MSTG1/2/3),were identified in the Y-linked area and considered to be strong candidate genes for sex determination or differentiation.Population genomic analysis showed that Guangdong accessions in China were genetically similar to Japanese accessions of mul-berry.In addition,genomic areas containing selective sweeps that distinguish domesticated mul-berry from wild populations in terms of flowering and disease resistance were identified.Our study provides an important genetic resource for sex identification research and molecular breeding in mulberry.

Objective:To investigate the value of texture analysis based on T 2WI and apparent diffusion coefficient (ADC) maps in discriminating low grade from high grade prostate cancer (PCa). Methods:Retrospective analysis was performed on patients who were confirmed to be PCa by pathology after surgery and underwent MRI examination in the department of radiology,Tongji Hospital,Tongji Medical College, Huazhong University of Science and Technology before radical surgery, including routine T 1WI, T 2WI and diffusion weighted imaging (DWI) sequences. 3D data analysis module of the MaZda software was used to manually draw region of interest (ROIs) slice by slice on T 2WI and ADC images, and generate volume of interest (VOI) of the entire tumor. MaZda software was also used to extract texture features. The independent sample t test or Mann-Whitney U test were used to identify the texture features with statistically significant differences between low and high grade PCa groups. Lasso regression model was used to select the best combination of texture features for identifying low and high grade PCa, and then the model was built. The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic performance of the model in both training cohort and test cohort. Results:The best texture feature combination selected by Lasso regression model were the S (1, 0, 0) correlation of T 2WI and the S (1, 0, 0) correlation, S (1, -1, 0) sum entropy and vertical-run length nonuniformity of ADC maps. The area under the ROC curve (AUC) of the model in training cohort was 0.823, and the sensitivity and specificity were 70.4% and 80.8%, respectively, which were better than the single texture feature. The AUC of the model in test cohort was 0.714, which was worse than training cohort. Conclusion:The texture analysis of T 2WI and ADC maps is valuable for the identification of low and high grade PCa.

Objective : To evaluate the safety and immunogenicity of a split-virion quadrivalent influenza vaccine. Methods : The healthy people aged three years or over in Wuyang County and Xiping County of Henan Province were divided into the experimental group, control group 1 and control group 2, and were vaccinated with split-virion quadrivalent influenza vaccines, split-virion trivalent influenza vaccines (without B/Victoria) and a split-virion trivalent influenza vaccines (without B/Yamagata) , respectively. The hemagglutination inhibition (HI) antibodies were detected before and after immunization. The incidence rate of adverse events following immunization (AEFI) , HI antibody positive conversion rate, the protection rate of HI antibodies and the growth of geometric mean titer (GMT) were calculated and compared with the standard of Food and Drug Administration (FDA). Results: Totally 2 924 people were recruited, with 975 in the experimental group, 974 in the control group 1 and 975 in control group 2. The incidence rate of AEFI in the experimental group was 11.7%, higher than 7.9% in control group 1 and 8.8% in control group 2 (P < 0.05) during 30 minutes and 8 days after inoculation. The positive conversion rates of HI antibodies of H1N1, H3N2, By and Bv in the experimental group were 78.5%, 53.3%, 78.3% and 62.9%, respectively. The rate differences of the positive conversion rates of HI antibodies of By between the experimental group and control group 2, and of Bv between the experimental group and control group 1 were 42.1% (95%CI: 38.0%-46.2%) and 33.2% (95%CI: 28.9%-37.5%) , with both lower limits of 95%CI more than -0.10. The GMT increase of HI antibodies was more than 2.5 times in the three groups. The protective rates of HI antibodies of H1N1, H3N2, By and Bv in the experimental group were 87.7%, 98.7%, 93.6% and 77.2%, respectively. The protective rates of HI antibodies of By in control group 2 and Bv in control group 1 were 71.1% and 51.0%, both lower than those in the experimental group (P < 0.05). Conclusions After the inoculation of the quadrivalent influenza vaccine, the positive conversion rates (>40%) , protection rates (>70%) and GMT increase (>2.5 times) of HI antibodies of H1N1, H3N2, By and Bv all meet the quality standards of FDA. The safety and immunogenicity of the quadrivalent influenza vaccine are not inferior to those of the trivalent influenza vaccine.

Objective To investigate the curative effect of ultra early enteral nutrition (EN) supplemented with probiotics in patients with severe acute pancreatitis (SAP).Methods Seventy-five SAP cases admitted in Second People's Hospital of Nantong from December 2014 to June 2018 were enrolled and assigned into 24 h group (who received EN & probiotics at 24 h,n =25),72 h group (who received EN & probiotics at 72 h,n =25) and the control group (who received EN & probiotics on day 7,n =25).After admission,the levels of PCT,hs-CRP and endotoxin on day 1,4 and 8 were detected.APACHE Ⅱ score on day 3 after admission,complication rate,infection rate,hospitalization duration and mortality were recorded.Results On day 1 after admission,serum PCT,hs-CRP and endotoxin concentrations among three groups were of no significant differences.On day 4 after admission,the serum concentrations of PCT and edotoxin in 24 h group were significantly lower than those in 72 h group and the control group [(3.12 ± 1.45) μg/L vs (5.26±1.52),(6.07 ± l.59) μg/L;(0.24 ± 0.02) EU/ml vs (0.35 ±0.03),(0.46±0.04) EU/ml].The serum hs-CRP in 24 h group was significantly lower than that of the control group [(20.71 ±4.89) μg/L vs (28.37 ± 4.64) μg/L],and the difference was statistically significant (P ＜ O.05).The serum concentrations of hs-CRP in 24 h group and 72 h group was of no significant difference.On day 7 after admission,the serum concentrations of PCT,hs-CRP and endotoxin in 24 h group were significantly lower than those in 72 h group and the control group (P ＜0.05);the serum concentrations of PCT,hs-CRP and endotoxin in 72 h group were significantly lower than those of the control group,and all the differences were statistically significant (all P ＜ 0.05).The score of APACHE Ⅱ (day 3 after admission) [(15.1 ± 1.8)],complication rate (52％),infection rate (8％),length of stay [(19.7 ± 5.3) d] in 24 h group were all significantly lower than those of 72h group [(17.0 ± 2.0),72％,32％,(25.4 ± 6.8) d] and control group [(18.3 ±2.6),84％,44％,(38.7 ± 12.6) d],and all the differences were statistically significant (all P ＜0.05).Conclusions Ultra EN supplemented with probiotics in treating the patients with SAP could significantly reduce the level of serum PCT,hs-CRP and endotoxin,decrease complication rate and mortality,and shorten the time of hospitalization.

Mitochondrial dysfunctions play major roles in ageing. How mitochondrial stresses invoke downstream responses and how specificity of the signaling is achieved, however, remains unclear. We have previously discovered that the RNA component of Telomerase TERC is imported into mitochondria, processed to a shorter form TERC-53, and then exported back to the cytosol. Cytosolic TERC-53 levels respond to mitochondrial functions, but have no direct effect on these functions, suggesting that cytosolic TERC-53 functions downstream of mitochondria as a signal of mitochondrial functions. Here, we show that cytosolic TERC-53 plays a regulatory role on cellular senescence and is involved in cognition decline in 10 months old mice, independent of its telomerase function. Manipulation of cytosolic TERC-53 levels affects cellular senescence and cognition decline in 10 months old mouse hippocampi without affecting telomerase activity, and most importantly, affects cellular senescence in terc cells. These findings uncover a senescence-related regulatory pathway with a non-coding RNA as the signal in mammals.

Objective To investigate the value of texture analysis based on MR ADC map of prostate in differentiating between low?grade and high?grade prostate cancer (PCa). Methods PCa confirmed by pathology after radical prostatectomy were analyzed retrospectively, all patients underwent multiparametric MRI before radical prostatectomy, including T1WI,T2WI and DWI. On the ADC map, ROI was drawn manually to encompass the whole tumor by ITK?SNAP software. The python?based pyradiomics package was used to extract 105 texture features. The intraclass correlation coefficient was used to evaluate the repeatability of the texture features. The independent sample t test or Mann?Whitney U test was used to exclude features that had no significant difference between low grade and high grade PCa. Lasso regression model and 5 fold cross validation method were used to obtain texture feature combination of the highest performance and develop a classification modelfor discriminating low from high grade PCa. ROC curve was used to evaluate the diagnostic efficiency of the model. Result Ninety patients with PCa confirmed by pathology after radical prostatectomywere analyzed retrospectively,including 36 patients with low?level PCa (GS≤3+4) and 54 patients with high?level PCa (GS≥4+3). The area under curve of the model was 0.841, with sensitivity 69.6% and specificity 91.2%, which was significantly higher than single texture feature or traditional mean ADC value. Conclusion Texture analysis based on MRI?ADC map of prostate could be used to discriminate low grade PCa from high grade PCa.

Objective: To investigate the value of texture analysis based on MR ADC map of prostate in differentiating between low-grade and high-grade prostate cancer (PCa). Methods: PCa confirmed by pathology after radical prostatectomy were analyzed retrospectively, all patients underwent multiparametric MRI before radical prostatectomy, including T₁WI,T₂WI and DWI. On the ADC map, ROI was drawn manually to encompass the whole tumor by ITK-SNAP software. The python-based pyradiomics package was used to extract 105 texture features. The intraclass correlation coefficient was used to evaluate the repeatability of the texture features. The independent sample t test or Mann-Whitney U test was used to exclude features that had no significant difference between low grade and high grade PCa. Lasso regression model and 5 fold cross validation method were used to obtain texture feature combination of the highest performance and develop a classification modelfor discriminating low from high grade PCa. ROC curve was used to evaluate the diagnostic efficiency of the model. Result: Ninety patients with PCa confirmed by pathology after radical prostatectomywere analyzed retrospectively,including 36 patients with low-level PCa (GS≤3+4) and 54 patients with high-level PCa (GS≥4+3). The area under curve of the model was 0.841, with sensitivity 69.6% and specificity 91.2%, which was significantly higher than single texture feature or traditional mean ADC value. Conclusion Texture analysis based on MRI-ADC map of prostate could be used to discriminate low grade PCa from high grade PCa.