The Chinese hamster ovary (CHO) cell is the most representative mammalian cell protein expression system, and it is widely used in recombinant protein, vaccine and other biopharmaceutical fields. However, due to its vulnerability to environmental factors, apoptosis, and metabolic inhibitors, CHO cells demonstrate poor robustness, and thus the integrated viable cell density and unit cell productivity are largely limited. To improve the robustness and foreign protein expression efficiency of CHO cells, we employed CRISPR/Cas9 to knock out the apoptosis genes Bax and Bak and the lactate dehydrogenase gene LDHa, thereby blocking apoptosis and lactic acid metabolism pathways. The results of apoptosis and single cell viability detection showed that the number of apoptotic cells in the knockout cell lines Bax^-/-, Bax-bak^-/-, and LDHa-Bax-bak^-/- was reduced by 22.51%, 37.73%, and 64.12%, respectively, compared with the wild-type cell line CHO-K1, which indicated that the anti-apoptotic ability was significantly improved. After staurosporine treatment, the single cell viability of Bax^-/-, Bax-bak^-/-, and LDHa-Bax-bak^-/- cells was increased by 30.8%, 22%, and 41.1%, respectively. After treatment with puromycin, the single cell viability of Bax^-/-, Bax-bak^-/-, and LDHa-Bax-bak^-/- cells was increased by 26.7%, 30.7%, and 38.8%, respectively. To further investigate the production performance of cells obtained after blocking apoptosis and lactic acid metabolism pathways, we induced transient expression of human tissue plasminogen activator (tPA) in these cells. The results showed that the secretion of tPA in Bax^-/-, Bax-Bak^-/-, and LDHa-Bax-Bak^-/- cells was 11.12%, 46.18%, and 63.13%, respectively, higher than that in wild-type CHO-K1 cells. The expression of intracellular tPA was increased by 35.65%, 130%, and 192.15%. In conclusion, blocking apoptosis and lactic acid metabolism pathways simultaneously can improve cell robustness and productivity, with the performance better than blocking the apoptosis pathway alone. The above results indicated that the constructed cell lines were expected to be the delivery carriers of protein drugs such as medicinal peptides, and better used for the treatment of diseases.
CHO Cells ; Cricetulus ; Animals ; Apoptosis/genetics* ; Lactic Acid/metabolism* ; Recombinant Proteins/biosynthesis* ; L-Lactate Dehydrogenase/genetics* ; bcl-2-Associated X Protein/genetics* ; bcl-2 Homologous Antagonist-Killer Protein/genetics* ; Cricetinae ; CRISPR-Cas Systems ; Staurosporine/pharmacology*

Objective To analyze the composition characteristics and biological functions of tumor cell subpopulations in glioblastoma(GBM)through multi-transcriptomics sequencing technology,and explore the hypoxia characteristics and spatial localization features of the mesenchymal-like(MES-like)tumor cell subpopulation in GBM and the influence on malignant biological behaviors.Methods Multi-transcriptomics sequencing data,including single-cell RNA sequencing(scRNA-seq)data(18 patients),bulk RNA sequencing(bulk RNA-seq)and spatial transcriptomics(ST)data of GBM,were employed to define cell subpopulations in GBM,and Gene Ontology(GO)and Gene Set Enrichment Analysis(GSEA)were utilized to analyze their functions.The proportions and locations of cell subpopulations in bulk RNA-seq data were evaluated with BayesPrism deconvolution.Immunofluorescence assay was conducted for verification on 12 paraffin samples of GBM from patients who visited the neurosurgical department of our hospital from 2015 to 2023 and met the pathological diagnostic criteria for GBM(10 males and 2 females,at an average age of 53.50 years and a median age of 54.50 years).pySCENIC was applied to predict specific transcription factors of tumor cell subpopulations.Results Tumor cells in GBM were highly heterogeneous,and could be mainly divided into 4 subpopulations:astrocyte-like(AC-like),neural progenitor-like(NPC-like),oligodendrocyte progenitor-like(OPC-like)and MES-like.Differential gene analysis found that the MES-like tumor cells highly expressed vascular endothelial growth factor A(VEGFA),adrenomedullin(ADM),N-myc downstream regulated 1(NDRG1),insulin like growth factor binding protein 5(IGFBP5),and A-kinase anchoring protein 12(AKAP12)(P<0.001).pySCENIC transcription factor prediction found that the high-active transcription factors of the MES-like tumor cells were AT-rich interaction domain 3A(ARID3A),FOS like 2,AP-1 transcription factor subunit(FOSL2),endothelial PAS domain protein 1(EPAS1),CCAAT enhancer binding protein delta(CEBPD),and CCAAT enhancer binding protein beta(CEBPB)(P<0.05).GO and GSEA enrichment analyses found that the MES-like tumor cells were enriched in hypoxia-related pathways,especially the pathway of cell responses to hypoxia levels(NES=2.437,P<0.001).BayesPrism deconvolution showed that the MES-like tumor cells mainly existed in PAN(Pseudopalisading cells around necrosis)and perinecrotic zone.Immunofluorescence assay confirmed CD44+(CD44 antigen)MES-like tumor cells were mainly located in hypoxia areas with highly expression of hypoxia inducible factor 1 subunit alpha(HIF1α)(P<0.01).Multivariate Cox regression analysis indicated that the MES-like tumor cells were significantly correlated with the adverse prognosis of GBM patients(HR=1.71,95%CI:1.38～2.11,P<0.001).Conclusion Tumor cells in GBM are of highly heterogeneity.They could be mainly divided into 4 subpopulations:AC-like,NPC-like,OPC-like and MES-like.MES-like tumor cells,mainly locating in PAN and perinecrotic zone,are characterized by hypoxia,which can promote the malignant progression of GBM.

Objective:To explore the effect of levonorgestrel-releasing intrauterine system (LNG-IUS) placed in the uterine cavity after transcervical preparation for the treatment of adenomyosis (AM).Methods:A retrospective study analyzed 219 cases of AM patients treated at Department of Obstetrics and Gynaecology, the Second Hospital of Hebei Medical University from March 2022 to February 2023. Among them, 112 cases were treated with LNG-IUS after hysteroscopy due to abnormal uterine bleeding or abnormal uterine echo suggested by gynecological ultrasound, and were recorded as the hysteroscopy+LNG-IUS group; 107 cases without the above manifestations were treated with LNG-IUS alone, and were recorded as the LNG-IUS group. The two groups were observed for dysmenorrhea severity, menstrual volume, endometrial thickness, anemia, serum CA125 levels, uterine volume, and adverse reactions such as abnormal vaginal bleeding and intrauterine contraceptive device displacement or shedding before and 1, 3, 6, and 12 months after treatment.Results:1) Postoperative pathology of hysteroscopy+LNG-IUS group after hysteroscopy included endometrial polyps [39.3% (44/112)], proliferative endometrium [16.1% (18/112)], uterine leiomyoma [11.6% (13/112)], secretory endometrium [8.9% (10/112)], irregular endometrial hyperplasia [6.2% (7/112)], simple hyperplasia [5.4% (6/112)], AM lesions [4.5% (5/112)], and others [8.0% (9/112)]. 2) The degree of dysmenorrhea and menstrual volume, as well as other indicators, improved after treatment in both groups (all P<0.001). 3) When comparing the two groups, the degree of dysmenorrhea, menstrual volume, endometrial thickness, anemia, and serum CA125 at 1, 3, 6, and 12 months of treatment, the hysteroscopy+LNG-IUS group was significantly better than those in the LNG-IUS group (all P<0.05), the difference of uterine volume at 12 months of treatment between the hysteroscopy+LNG-IUS group [(109.76±32.90) cm 3] and the LNG-IUS group [(120.84±31.30) cm 3] was statistically significant ( P=0.011). 4) The cumulative incidence of adverse reactions of the two groups was statistically significant ( P=0.001) between the hysteroscopy+LNG-IUS group [24.1% (27/112)] and the LNG-IUS group [52.3% (56/107), P=0.001]. The hysteroscopy+LNG-IUS group had lower rates of vaginal irregular bleeding [8.0% (9/112)] and ring displacement or shedding [3.6% (4/112)] than those in the LNG-IUS group [23.4% (25/107), P=0.002; 11.2% (12/107), P=0.030]. Conclusion:After treatment with hysteroscopy, the placement of LNG-IUS and the simple intrauterine placement of LNG-IUS both significantly improve the severity of dysmenorrhea, endometrial thickness, anemia, menstrual volume, CA125, and uterine volume. The treatment effect of the LNG-IUS group after hysteroscopy is better and adverse reactions are milder, and it is expected to become the preferred option for the long-term management of conservative treatment of AM.

Suppressor of cytokine signaling 3(SOCS3),a member of SOCS family,plays a major role in cell signaling trans-duction.SOCS3 is important in regulation of immune homeostasis and also involved in the control of energy metabolism,mainly through regulating JAK/STAT pathway.Recent studies have shown that SOCS3 was closely associated with glycolipid metabolic diseases like diabetes and obesity,and can be used as a new target for treatment of these glycolipid metabolic diseases.This review focuses on the latest research progress of the role and mechanism of SOCS3 in glycolipid metabolic diseases,and discusses the possibility of SOCS3 as a therapeutic target for glycolipid metabolic diseases.

Objective:To explore the effect of levonorgestrel-releasing intrauterine system (LNG-IUS) placed in the uterine cavity after transcervical preparation for the treatment of adenomyosis (AM).Methods:A retrospective study analyzed 219 cases of AM patients treated at Department of Obstetrics and Gynaecology, the Second Hospital of Hebei Medical University from March 2022 to February 2023. Among them, 112 cases were treated with LNG-IUS after hysteroscopy due to abnormal uterine bleeding or abnormal uterine echo suggested by gynecological ultrasound, and were recorded as the hysteroscopy+LNG-IUS group; 107 cases without the above manifestations were treated with LNG-IUS alone, and were recorded as the LNG-IUS group. The two groups were observed for dysmenorrhea severity, menstrual volume, endometrial thickness, anemia, serum CA125 levels, uterine volume, and adverse reactions such as abnormal vaginal bleeding and intrauterine contraceptive device displacement or shedding before and 1, 3, 6, and 12 months after treatment.Results:1) Postoperative pathology of hysteroscopy+LNG-IUS group after hysteroscopy included endometrial polyps [39.3% (44/112)], proliferative endometrium [16.1% (18/112)], uterine leiomyoma [11.6% (13/112)], secretory endometrium [8.9% (10/112)], irregular endometrial hyperplasia [6.2% (7/112)], simple hyperplasia [5.4% (6/112)], AM lesions [4.5% (5/112)], and others [8.0% (9/112)]. 2) The degree of dysmenorrhea and menstrual volume, as well as other indicators, improved after treatment in both groups (all P<0.001). 3) When comparing the two groups, the degree of dysmenorrhea, menstrual volume, endometrial thickness, anemia, and serum CA125 at 1, 3, 6, and 12 months of treatment, the hysteroscopy+LNG-IUS group was significantly better than those in the LNG-IUS group (all P<0.05), the difference of uterine volume at 12 months of treatment between the hysteroscopy+LNG-IUS group [(109.76±32.90) cm 3] and the LNG-IUS group [(120.84±31.30) cm 3] was statistically significant ( P=0.011). 4) The cumulative incidence of adverse reactions of the two groups was statistically significant ( P=0.001) between the hysteroscopy+LNG-IUS group [24.1% (27/112)] and the LNG-IUS group [52.3% (56/107), P=0.001]. The hysteroscopy+LNG-IUS group had lower rates of vaginal irregular bleeding [8.0% (9/112)] and ring displacement or shedding [3.6% (4/112)] than those in the LNG-IUS group [23.4% (25/107), P=0.002; 11.2% (12/107), P=0.030]. Conclusion:After treatment with hysteroscopy, the placement of LNG-IUS and the simple intrauterine placement of LNG-IUS both significantly improve the severity of dysmenorrhea, endometrial thickness, anemia, menstrual volume, CA125, and uterine volume. The treatment effect of the LNG-IUS group after hysteroscopy is better and adverse reactions are milder, and it is expected to become the preferred option for the long-term management of conservative treatment of AM.

Suppressor of cytokine signaling 3(SOCS3),a member of SOCS family,plays a major role in cell signaling trans-duction.SOCS3 is important in regulation of immune homeostasis and also involved in the control of energy metabolism,mainly through regulating JAK/STAT pathway.Recent studies have shown that SOCS3 was closely associated with glycolipid metabolic diseases like diabetes and obesity,and can be used as a new target for treatment of these glycolipid metabolic diseases.This review focuses on the latest research progress of the role and mechanism of SOCS3 in glycolipid metabolic diseases,and discusses the possibility of SOCS3 as a therapeutic target for glycolipid metabolic diseases.

Objective:To investigate the curative effects of Shugan Hewei Zhitong Recipe on chronic gastritis complicated by Helicobacter pylori infection. Methods:Eighty-seven patients with chronic gastritis complicated by Helicobacter pylori infection who received treatment in the Huguosi Hospital of Traditional Chinese Medicine, Beijing University of Chinese Medicine, China from July 2018 to June 2020 were included in this study. They were randomly assigned to undergo treatment with either conventional Western medicine ( n = 43, control group) or Shugan Hewei Zhitong Recipe based on conventional Western medicine ( n = 44, observation group) for 2 weeks. Before and after treatment, traditional Chinese medicine syndrome scores were compared between the two groups. Curative effects of Shugan Hewei Zhitong Recipe on chronic gastritis complicated by Helicobacter pylori infection were evaluated. Results:Total response rate in the observation group was significantly higher than that in the control group [91% (40/44) vs. 72% (31/43), χ2 = 5.13, P = 0.024]. In each group, the traditional Chinese medicine symptom score post-treatment was significantly lower than the traditional Chinese medicine symptom score pre-treatment (both P < 0.001). After treatment, the traditional Chinese medicine score of the observation group was significantly lower than that of the control group ( P < 0.05). There were no adverse reactions in each group. Conclusion:Shugan Hewei Zhitong Recipe is safe and effective on chronic gastritis complicated by Helicobacter pylori infection, which is worthy of clinical promotion.

OBJECTIVE To compare the dosage forms and specifications of over-the-counter drug （OTC）（chemical drugs ） for children at home and abroad ，and to provide reference for the addition of new dosage forms and specifications of OTC for children in China . METHODS Data analysis was used to comb the active ingredients of OTC single -ingredient preparation for children in China . The similarities and differences of the dosage forms and specifications of OTC for children with the same active ingredients among China and 8th edition of WHO Model List of Essential Medicines for Children （WHO EMLc ）and US/UK/EU （this article refers specifically to EU countries ）/Japan were analyzed by comparative analysis . RESULTS There were 72 active ingredients of OTC single -ingredient preparation for children in China ，corresponding to 34 dosage forms and 216 specifications； 39 same active ingredients of OTC for children were retrieved in WHO EMLc and US/UK/EU/Japan ，corresponding to 38 dosage forms and 258 specifications. Among OTC for children corresponding to 39 active ingredients ，there were 10 unique dosage forms in China ，and 16 unique dosage forms in WHO EMLc and US/UK/EU/Japan ，of which some dosage forms have advantages for children（such as chewing gums ，gels for external use ，spray for oral liquid ，etc.），were included in the latter while not included in China . There were 107 unique specifications in China ，and 214 unique specifications in WHO EMLc and US/UK/EU/Japan ，of which the division of applicable age groups for specifications was more detailed . In addition ，the dosage forms and specifications corresponding to a few active ingredients （such as ibuprofen ，cetirizine hydrochloride ）were not included in OTC for children in China， while included in UK/EU . CONCLUSIONS The dosage forms and specifications of OTC for children in WHO EMLc and US/UK/EU/Japan are generally more abundant and E-mail：zjfyouyou@163.com flexible than in China . Relevant departments in China should learn from the advanced experience of WHO and foreign countries and increase the research and development of dosage forms and specifications of OTC for children .

Objective:To summarize our experience in prevention of COVID-19 infection in emergency and confined operations during the first 3 weeks after Spring Festival in 2020.Methods:From February 3rd to 23rd, 2020, 151 patients were admitted to Department of Orthopaedic Trauma, Beijing Jishuitan Hospital for emergency and confined operations. In this cohort, 125 patients were admitted to ordinary wards. They were 70 males and 55 females with an age of 51.1 years ± 14.9 years. Of them, 2 were subjected to emergency operation and 123 to confined operation. The mean time from injury to operation was 9.9 days ± 6.1 days. There were 26 cases in the senile wards, 7 males and 19 females with an age of 80.8 years ± 7.0 years all of whom underwent confined operations. The mean time from injury to operation was 8.4 days ± 6.3 days. The protocols for emergency diagnosis, admission, emergency and confined operations, postoperative rehabilitation and management of suspects with COVID-19 during the epidemic of COVID-19 were optimized according to Diagnosis and Treatment Protocols for Novel Coronavirus Pneumonia (Trial version 5), emergency responding pre-plans of our hospital, and our experience in Enhanced Recovery After Surgery (ERAS) as well.Results:The patients in the ordinary wards had a hospital stay of 6.8 days ± 4.6 days while those in the senile wards 5.1 days ± 2.0 days. Abnormal temperature (≥37.3 ℃) was observed perioperatively in 17 cases in the ordinary wards. It was absorption fever in all and appeared in 4 cases upon admission. Fever appeared in 11 patients in the senile wards and upon admission in 3 of them. One senile patient who had been diagnosed of normal pneumonia returned to normal temperature and remained stable conditions after antibiotic therapy. The other patients were free of complications related to COVID-19 during their hospital stay.Conclusion:The first-line medical staff working at emergency department, wards and surgical theaters must heighten their vigilance against COVID-19 infection and rigorously follow protocols for prevention of COVID-19 infection in their daily clinical practice.

Objective To investigate the needs and service of rehabilitation status of the people with intellectual disability, to support rehabilitation service for them. Methods A total of 167,872 cases with intellectual disability from the Administration Data of 2017 Basic Service Status and Needs of People with Disabilities in Henan Province were analyzed and rehabilitation needs included medical service and support, assistive productions, and rehabilitation training. Results 56.4% of the people with intellectual disability reported their rehabilitation needs. The rehabilitation needs were different with their ages, levels of disability and household registration (χ2 > 976.42, P < 0.001). Only 19.0% of the people with intellectual disability accessed rehabilitation services, and the influential factors included demographic variables, family economic status, transportation, knowledge and information, etc. Conclusion There are huge needs of rehabilitation services for the people with intellectual disability needed to be tailored. It is necessary to increase the supply of rehabilitation services using approach of precise rehabilitation, and improve the quality of rehabilitation services.