Chinese hamster ovary (CHO) cells are the most established and versatile mammalian expression system for the large-scale production of recombinant therapeutic proteins, owing to their genetic stability, adaptability to serum-free suspension culture, and ability to perform human-like post-translational modifications. More than 70% of biologics approved by the U.S. Food and Drug Administration rely on CHO-based production platforms, underscoring their central role in modern biopharmaceutical manufacturing. Despite these advantages, CHO systems continue to face three persistent bottlenecks that limit their potential for high-yield, reproducible, and cost-efficient production: excessive metabolic burden during high-density culture, heterogeneity of glycosylation patterns, and progressive loss of long-term expression stability. This review provides an integrated analysis of recent advances addressing these challenges and proposes a forward-looking framework for constructing intelligent and sustainable CHO cell factories. In terms of metabolic regulation, excessive lactate and ammonia accumulation disrupts energy balance and reduces recombinant protein synthesis efficiency. Optimization of culture parameters such as temperature, pH, dissolved oxygen, osmolarity, and glucose feeding can effectively alleviate metabolic stress, while supplementation with modulators including sodium butyrate, baicalein, and S-adenosylmethionine promotes specific productivity (qP) by modulating apoptosis and chromatin structure. Furthermore, genetic engineering strategies—such as overexpression of MPC1/2, HSP27, and SIRT6 or knockout of Bax, Apaf1, and IGF-1R—have demonstrated significant improvements in cell viability and product yield. The combination of multi-omics metabolic modeling with artificial intelligence (AI)-based prediction offers new opportunities for building self-regulating CHO systems capable of dynamic adaptation to environmental stress. Regarding glycosylation uniformity, which determines therapeutic efficacy and immunogenicity, gene editing-based glycoengineering (e.g., FUT8 knockdown or ST6Gal1 overexpression) has enabled the humanization of CHO glycan profiles, minimizing non-human sugar residues and enhancing drug stability. Process-level strategies such as galactose or manganese co-feeding and fine control of temperature or osmolarity further allow rational regulation of glycosyltransferase activity. Additionally, in vitro chemoenzymatic remodeling provides a complementary route to construct human-type glycans with defined structures, though industrial applications remain constrained by cost and scalability. The integration of model-driven process design and AI feedback control is expected to enable real-time prediction and correction of glycosylation deviations, ensuring batch-to-batch consistency in continuous biomanufacturing. Long-term expression stability, another critical challenge, is often impaired by promoter silencing, chromatin condensation, and random genomic integration. Molecular optimization—such as the use of improved promoters (CMV, EF-1α, or CHO endogenous promoters), Kozak and signal peptide refinement, and incorporation of chromatin-opening elements (UCOE, MAR, STAR)—helps maintain durable transcriptional activity, while site-specific integration systems including Cre/loxP, Flp/FRT, φC31, and CRISPR/Cas9 can enable single-copy, position-independent gene insertion at genomic safe-harbor loci, ensuring stable, predictable expression. Collectively, this review highlights a paradigm shift in CHO system optimization driven by the convergence of genome editing, synthetic biology, and artificial intelligence. The transition from empirical optimization to rational, data-driven design will facilitate the development of programmable CHO platforms capable of autonomous regulation of metabolic flux, glycosylation fidelity, and transcriptional activity. Such intelligent cell factories are expected to accelerate the transformation from laboratory-scale research to industrial-scale, high-consistency, and economically sustainable biopharmaceutical manufacturing, thereby supporting the next generation of efficient and customizable biologics manufacturing.

Chinese hamster ovary (CHO) cells are the most established and versatile mammalian expression system for the large-scale production of recombinant therapeutic proteins, owing to their genetic stability, adaptability to serum-free suspension culture, and ability to perform human-like post-translational modifications. More than 70% of biologics approved by the U.S. Food and Drug Administration rely on CHO-based production platforms, underscoring their central role in modern biopharmaceutical manufacturing. Despite these advantages, CHO systems continue to face three persistent bottlenecks that limit their potential for high-yield, reproducible, and cost-efficient production: excessive metabolic burden during high-density culture, heterogeneity of glycosylation patterns, and progressive loss of long-term expression stability. This review provides an integrated analysis of recent advances addressing these challenges and proposes a forward-looking framework for constructing intelligent and sustainable CHO cell factories. In terms of metabolic regulation, excessive lactate and ammonia accumulation disrupts energy balance and reduces recombinant protein synthesis efficiency. Optimization of culture parameters such as temperature, pH, dissolved oxygen, osmolarity, and glucose feeding can effectively alleviate metabolic stress, while supplementation with modulators including sodium butyrate, baicalein, and S-adenosylmethionine promotes specific productivity (qP) by modulating apoptosis and chromatin structure. Furthermore, genetic engineering strategies—such as overexpression of MPC1/2, HSP27, and SIRT6 or knockout of Bax, Apaf1, and IGF-1R—have demonstrated significant improvements in cell viability and product yield. The combination of multi-omics metabolic modeling with artificial intelligence (AI)-based prediction offers new opportunities for building self-regulating CHO systems capable of dynamic adaptation to environmental stress. Regarding glycosylation uniformity, which determines therapeutic efficacy and immunogenicity, gene editing-based glycoengineering (e.g., FUT8 knockdown or ST6Gal1 overexpression) has enabled the humanization of CHO glycan profiles, minimizing non-human sugar residues and enhancing drug stability. Process-level strategies such as galactose or manganese co-feeding and fine control of temperature or osmolarity further allow rational regulation of glycosyltransferase activity. Additionally, in vitro chemoenzymatic remodeling provides a complementary route to construct human-type glycans with defined structures, though industrial applications remain constrained by cost and scalability. The integration of model-driven process design and AI feedback control is expected to enable real-time prediction and correction of glycosylation deviations, ensuring batch-to-batch consistency in continuous biomanufacturing. Long-term expression stability, another critical challenge, is often impaired by promoter silencing, chromatin condensation, and random genomic integration. Molecular optimization—such as the use of improved promoters (CMV, EF-1α, or CHO endogenous promoters), Kozak and signal peptide refinement, and incorporation of chromatin-opening elements (UCOE, MAR, STAR)—helps maintain durable transcriptional activity, while site-specific integration systems including Cre/loxP, Flp/FRT, φC31, and CRISPR/Cas9 can enable single-copy, position-independent gene insertion at genomic safe-harbor loci, ensuring stable, predictable expression. Collectively, this review highlights a paradigm shift in CHO system optimization driven by the convergence of genome editing, synthetic biology, and artificial intelligence. The transition from empirical optimization to rational, data-driven design will facilitate the development of programmable CHO platforms capable of autonomous regulation of metabolic flux, glycosylation fidelity, and transcriptional activity. Such intelligent cell factories are expected to accelerate the transformation from laboratory-scale research to industrial-scale, high-consistency, and economically sustainable biopharmaceutical manufacturing, thereby supporting the next generation of efficient and customizable biologics manufacturing.

Objective: To investigate the trend in disease burden of asthma attributable to tobacco in China from 1990 to 2021, so as to provide the basis for improving intervention measures of asthma. Methods: Data on asthma-related mortality and disability-adjusted life years (DALY) attributable to tobacco among adults aged ≥30 years in China from 1990 to 2021 were collected from the Global Burden of Disease (GBD) 2021 database. Age-standardized mortality and age-standardized DALY rate were calculated using the GBD world standard population structure to analyze the tobacco-attributable asthma burden. The average annual percent change (AAPC) was employed to evaluate temporal trends in the age-standardized mortality and DALY rate from 1990 to 2021. Results: In China, the age-standardized mortality and age-standardized DALY rate of asthma attributable to tobacco decreased from 0.73/100 000 and 22.20/100 000 in 1990 to 0.17/100 000 and 6.64/100 000 in 2021, showing downward trends (AAPC=-4.603% and -3.888%, both P<0.05). Among males, the tobacco-attributable age-standardized mortality and age-standardized DALY rate declined from 1.44/100 000 and 41.05/100 000 in 1990 to 0.36/100 000 and 12.79/100 000 in 2021 (AAPC=-4.369% and -3.810%, both P<0.05). Among females, the corresponding rates decreased from 0.21/105 and 5.37/105 to 0.03/105 and 1.08/105 (AAPC=-6.074% and -5.074%, both P<0.05). In 2021, males had higher tobacco-attributable age-standardized mortality and age-standardized DALY rate for asthma than females. Both the mortality and DALY rate of asthma attributable to tobacco increased with age, peaking in the age group ≥80 years at 7.84/100 000 and 112.07/100 000, respectively. Conclusion From 1990 to 2021, the disease burden of asthma attributable to tobacco showed a declining trend in China, with males and elderly population aged ≥80 years bearing a relatively heavier disease burden.

OBJECTIVE To investigate the clinical characteristics of euglycemic diabetic ketoacidosis （euDKA） induced by empagliflozin， and provide references for safe medication. METHODS Case reports of euDKA induced by empagliflozin were retrieved and analyzed from PubMed， Web of Science， CNKI， and Wanfang Data. The Naranjo’s scale was adopted to assess the causal relationship between empagliflozin and euDKA. RESULTS A total of 29 patients were enrolled， including 16 males and 13 females，with a median age of 58 years. All patients were diagnosed with type 2 diabetes mellitus； 22 patients had at least one comorbidity. The median time to euDKA onset was 60 days after administration， and 7 cases occurred within 2 weeks of treatment. Perioperative periods，fasting，and low-carbohydrate or ketogenic diets were the main predisposing factors. Common presentations included nausea，vomiting， fatigue， tachypnea，and tachycardia. The causal relationship between empagliflozin and euDKA was assessed as “probable” in all cases. According to adverse reaction severity grading， 13 cases were grade 3 （severe） and 16 cases were grade 4 （life-threatening）. All patients discontinued empagliflozin and recovered after corresponding treatment. CONCLUSIONS euDKA is a serious adverse reaction to empagliflozin， which mostly occurs early in treatment. Perioperative periods， fasting， and ketogenic diets are prone to induce euDKA. Elderly patients and those with comorbidities are at higher risk. Risk assessment should be conducted before clinical administration； blood ketones and acid-base monitoring should be strengthened in the early medication period； withhold the drug before elective surgery， avoid prolonged fasting；upon diagnosis， discontinue empagliflozin and initiate intravenous insulin， provide fluid resuscitation promptly， and use other treatments to improve patient prognosis.

ObjectiveTo evaluate the impact of narrative medicine education on the narrative ability of resident physicians undergoing standardized residency training， and to explore its application value in clinical practice. MethodsA total of 23 obstetricians and gynecologists who participated in residency training at the Fourth Hospital of Hebei Medical University from October 2021 to June 2024 were randomly selected to receive a 3-month residency training program integrated with narrative medicine education， including narrative theory learning， text reading， reflective writing， and scenario-based case analysis. A questionnaire survey was conducted to analyze the personal situation of resident physicians， their narrative ability before and after receiving narrative medicine education， and their satisfaction with teaching. ResultsThe results of the questionnaire survey showed that resident physicians who had received narrative medicine education scored higher on the narrative ability assessment scale than before training， including improved narrative abilities in the dimensions of life and health narrative awareness， professional narrative thinking， professional development narrative behavior， peer communication narrative behavior， and doctor-patient interaction narrative behavior （P<0.05）. However， there were no statistically significant differences in the dimensions of life and health narrative behavior and family connection narrative behavior （P>0.05）. Meanwhile， resident physicians’ interest in active learning， clinical thinking ability， doctor-patient communication ability， and satisfaction with teaching methods have also been improved （P<0.05）. ConclusionNarrative medicine education can effectively enhance the narrative ability of resident physicians and make up for the current deficiencies in humanistic literacy and ethical education in current medical education. It is of great significance for improving doctor-patient relationships and the quality of medical services. Therefore， it is recommended to integrate narrative medicine education into the regular training curriculum for resident physicians.

Objective To develop an ultra high performance liquid chromatography-mass spectrometry(UPLC-MS)method for quantifying serum levels of norepinephrine(NE)and 5-hydroxytryptamine(5-HT),and to monitor the dynamic changes in these neurotransmitters during the process of establishing a model of acute reserpine-induced depression-like mice.Methods By evaluating matrix effect,recovery,precision,and accuracy efficiency,an UPLC-MS method for determining the concentrations of NE and 5-HT in serum was established.Forty-eight C57 mice were randomly divided into normal control and model groups,which were intraperitoneally injected with physiological saline(10 mL/kg)and reserpine(2.5 mg/kg),respectively.At various time points after intraperitoneal injection,the degree of ptosis and decreases in body temperature of the mice were observed before orbital blood was sample for the determination of NE and 5-HT levels.Results The concentrations of NE and 5-HT showed good linearity within the range of 15.63 to 2000.00 ng/mL,with R2 values greater than 0.999.The results of methodological validation met the requirements for the analysis of biological samples,with a lower limit of quantification of 15.63 ng/mg.After intraperitoneal injection of reserpine,the model mice exhibited varying body temperature decreases and ptosis.At 1 and 2 h post-administration,the depression-like symptoms in the model group were significantly different from those of the normal control group(P＜0.01).The body temperature of mice in the model group was significantly lower than that of mice in the normal control group(P＜0.01),while the score of the eyelid ptosis was significantly higher(P＜0.001).The levels of NE and 5-HT in the serum of model mice were also significantly depleted,and were significantly different from those of the normal control group at 0.5,1 and 2 h(P＜0.05).Conclusion The study process of established a rapid and accurate method for dynamically observing the changes in NE and 5-HT levels during the process of establishing a model of acute reserpine-induced depression-like mice,which might contribute to the study of the pathogenesis of depression and the development of new antidepressant drugs.

ObjectiveTo investigate the serotype distribution and antimicrobial resistance characteristics of foodborne Salmonella in infectious diarrhea cases in Minhang District, Shanghai, and to provide evidence for clinical diagnosis, treatment, prevention, and control of salmonellosis. MethodsFecal or anal swab samples were collected from foodborne diarrhea cases at sentinel hospitals between 2019 and 2023 in Minhang District of Shanghai. Salmonella was isolated, biochemically identified, and serotyped. Antimicrobial susceptibility testing was performed via the microbroth dilution method. ResultsA total of 4 294 samples were collected, from which 224 Salmonella strains were isolated, with an isolation rate of 5.22%. There was no gender difference in the positive rate of Salmonella. The positive rate showed seasonal fluctuations (χ²=160.475, P<0.001), with a higher positive rate in summer and autumn than in spring and autumn. Among different occupational groups, the positive rate was the highest among retired people (7.30%) (χ²=20.023, P=0.001). Among different age groups, the positive rate was the highest among those aged 80 years and above (7.33%) (χ²=13.491, P=0.009). A total of 28 serotypes were identified, with Salmonella Enteritidis being the most predominant, followed by Salmonella Typhimurium. Among them, 154 strains were multi-drug resistant to three or more types of antibiotics (68.75%). The most common drug resistance patterns among multi-drug resistant strains were ampicillin (AMP)-ampicillin/sulbactam (AMS)-sulfamethoxazole/trimethoprim (SXT)-chloramphenicol (CHL)-tetracycline (TET)-streptomycin (STR), AMP-TET-STR and AMP-AMS-TET-STR, with 9 strains being detected in each isolates. ConclusionThe positive rate of foodborne Salmonella in Minhang District, Shanghai, is high in summer and autumn, with a higher infection rate among retired people and those aged 60 years and above being the high-risk group. It is recommended to strengthen the protection for high-risk groups, implement targeted prevention and control measures in summer and autumn to reduce the risk of infection, and pay attention to guiding clinical medication and infection control.

Objective:To investigate the distribution of CGG repeat numbers in the FMR1 gene among reproductive-age women in China, providing data reference for carrier screening and genetic counseling of Fragile X syndrome. Methods:This cross-sectional study recruited 16 610 reproductive-age women from 12 medical institutions between July 2022 and October 2023. Peripheral venous blood samples (3 mL) were collected, and genomic DNA was extracted. The number of CGG repeats in the FMR1 gene was determined using the triplet-primed polymerase chain reaction (TP-PCR) combined with capillary electrophoresis technology. Statistical analyses were performed to assess the prevalence and distribution of CGG repeat expansions. Results:Among 16 610 women of childbearing age, 5 684 (34.220%) women had the same number of CGG repeats in the two alleles of FMR1 gene, and 10 926 (65.780%) women had different numbers of repeats in the two alleles. Among the 33 220 FMR1 alleles in 16 610 women of reproductive age, the most common CGG repeat numbers were 29 [48.645% (16 160/33 220)] and 30 [26.276% (8 729/33 220)], while the most frequent CGG genotype was CGG 29/29 [24.726% (4 107/16 610)]. The CGG repeat numbers of FMR1 gene were normal in 16 498 women (99.326%). Among the 112 women (0.674%) with CGG repeat abnormities, 96 (0.578%) women were classified as intermediate carriers, 15 (0.090%) as premutation carriers, and 1 (0.006%) as a full mutation carrier, whose CGG genotype was (36, >200). Conclusion:In the general reproductive-age female population in China, the normal CGG repeat numbers of the FMR1 gene account for 99.326%, while the intermediate carrier rate is 0.578%, and the combined carrier rate of the premutation and full mutation types is 0.096%.

Objective To explore clinical effect of accessory scaphoid bone fusion in treating type Ⅱ painful accessory scaphoid bone.Methods A retrospective analysis was performed on 26 patients with type Ⅱ painful accessory navicular bone treated by accessory navicular bone fusion from January 2012 to June 2022,including 1 male and 25 females,aged from 18 to 70 years old with an average of(44.61±16.32)years old;10 patients with type Ⅱ A and 16 patients with type Ⅱ B;20 patients with simple fusion and 6 patients with fusion plus calcaneal translocation osteotomy.Changes of Meary angle,Pitch angle,an-teroposterior talar-first metatarsal angle(T1MA),talonavicular coverage angle(TCA),lateral talocalcaneal angle(LTCA)be-fore operation and 6 months after operation were observed and compared,and American Orthopedic Foot and Ankle Society(AOFAS)foot and ankle score and visual analogue scale(VAS)were used to explore clinical effect.Results All 26 patients were followed up for 7 to 24 months with an average of(10.72±3.94)months.Meary angle,Pitch angle,T1MA,TCA and LTCA were improved from(9.20±2.57)°,(16.45±3.57)°,(33.34±5.02)°,(22.42±5.86)°,(48.89±4.43)° before opertaion to(3.33±1.06)°,(22.33±4.56)°,(23.89±3.48)°,(11.83±2.67)°,(36.50±3.50)° at 6 months after operation,the difference were statistically significant(P＜0.01).Postoperative AOFAS foot and ankle score were(86.24±4.33)and(93.18±6.02)for type Ⅱ A and type Ⅱ B at 6 months,which were significantly improved compared with those for type Ⅱ A and type Ⅱ B before op-eration(67.34±6.55)and(65.12±9.51),and the difference was statistically significant(P＜0.01);20 patients got excellent re-sult,5 good and 1 poor.Preoperative VAS of type ⅡA(5.67±1.58)and type Ⅱ B(5.77±1.49)were improved to(2.13±1.01)and(1.43±0.68)at 6 months after operation,with statistical significance(P＜0.01).Conclusion Fusion of accessory navicular bone in patients with type Ⅱ painful accessory navicular bone combined with internal calcaneal osteotomy in patients with par-tial calcaneal valvaration could effectively correct flat foot deformity and relieve pain,and could be used as a clinical treatment for painful accessory navicular bone.

Hepatocellular carcinoma(HCC),as the most common type of liver cancer,poses a significant threat to global public health due to its high incidence and mortality rates.This paper delves into the etiology,pathogenesis,and syndrome differentiation of liver cancer from the perspective of"dispersing qi and fortifying the body resistance",based on the clinical experience of renowned traditional Chinese medicine(TCM)practitioner,Prof.Changquan Ling.Prof.Ling believes that the development of liver cancer is closely related to the disruption of liver qi flow,the accumulation of blood stasis over time,and the generation of toxin from long-term stagnation,accompanied by pathological changes such as imbalance of yin and yang,deficiency of the body's vital qi and accumulation of pathogenic factors,and internal blazing of cancer toxins.In terms of treatment,he emphasizes the principles of dispersing qi and fortifying the body resistance,addressing both the root cause and symptoms.This is achieved by regulating the functions of viscera,improving the stagnation of qi flow,and supplemented by methods such as clearing heat and detoxifying,and softening and dispersing hard masses,aiming to break the vicious cycle of qi stagnation,deficiency of vital qi,and pathogenic factor generation,thereby promoting the recovery from the disease.Through detailed analysis of clinical cases,this paper demonstrates Prof.Ling's unique insights and significant efficacy in treating liver cancer through"dispersing qi"to"fortify the body resistance",ultimately achieving"tumor suppression".This provides new references and perspectives for the clinical diagnosis and treatment of liver cancer in TCM.