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.

ObjectiveZheng’s San Qi San (ZSQS) power, a classic traditional Chinese medicine (TCM) formula, is used for treating soft tissue injuries involving muscles, tendons, and ligaments. However, its underlying therapeutic mechanisms remain unclear. This study aimed to screen and identify pharmaceutically active ingredients and their candidate biomolecule targets, and further elucidate the molecular mechanism of ZSQS in the treatment of skeletal muscle injury. MethodsNetwork pharmacology was employed to construct “ZSQS-component-target”, “protein-protein interaction (PPI)” and “active ingredient-core protein-pathway” networks to predict the key active ingredients and potential core targets of ZSQS for skeletal muscle injury. The predicted results were then validated via microarray data from the GEO database. Molecular docking was then performed to assess the binding ability between the screened active ingredients of ZSQS and the candidate core targets. Moreover, liquid chromatography-mass spectrometry (LC-MS) was used for qualitative and quantitative analysis to verify the active components of the drug and ZSQS serum. Finally, an animal model of eccentric exercise-induced skeletal muscle injury and a myotube cell model of oxidative stress-induced injury were established to validate the effects of ZSQS and its interventional effects on the biological functions of critical targets, thereby demonstrating the potential therapeutic mechanism of ZSQS. ResultsAmong the 111 active components identified in ZSQS and their corresponding 204 targets related to the skeletal muscle injury repair process, 14 core targets (including AKT1) and 4 core active components (quercetin, luteolin, kaempferol, and β‑sitosterol) were screened out, while the corresponding metabolites of quercetin, luteolin and kaempferol were detected in the ZSQS serum. Among these targets, 5 candidate genes (IL-6, CASP3, HIF1A, STAT3, and JUN) overlapped with the differential expression screening results with GEO data, and IL-6 was confirmed to be enriched in the PI3K/AKT pathway. Combined with the prediction results of the AKT expression levels, these findings suggest that the phosphorylation level of AKT1 plays a core role in the therapeutic mechanism of ZSQS. Molecular docking analysis further revealed that the PH domain of AKT1 had high binding energy with all 4 core active components, as verified by LC-MS. Finally, animal model studies have shown the promoting effect of ZSQS administration on skeletal muscle injury repair and its possible antioxidant damage mechanism. Cell model studies further demonstrated that ZSQS-containing serum, core active ingredient combination therapy, and quercetin monomer could increase the phosphorylation level of AKT, promote the nuclear translocation of Nrf2, upregulate the expression of downstream antioxidant enzymes (SOD, GPx, and GR), and inhibit the expression of inflammatory factors (IL-6 and TNF-α), thereby alleviating oxidative stress and the inflammatory response. ConclusionZSQS alleviates skeletal muscle injury mainly by activating the AKT/Nrf2 signaling pathway, enhancing cellular antioxidant and anti-inflammatory capabilities. The results of this study provide a scientific basis for the clinical application and modernized development of ZSQS.

ObjectiveTo investigate the association between insulin resistance （IR） and cardiovascular metabolic indicators， including blood pressure， blood lipids， and serum uric acid， in a middle-aged population. MethodsBased on the baseline survey of the Hong Sifang cohort of China National Salt Industry Group， a total of 575 participants were included. According to the homeostasis model assessment of insulin resistance， the participants were divided into an IR group （n=116） and a non-IR group （n=459）. Differences between the two groups were compared using the t-test and chi-square test. Multivariate logistic regression analysis was performed to evaluate the associations between IR and abnormalities in metabolic indicators. ResultsA total of 575 participants were included， including 390 men and 185 women， with a mean age of （48.58 ± 5.14） years and a mean body mass index （BMI） of （24.16 ± 2.97） kg/m². Blood pressure， serum uric acid， and blood lipid levels were significantly higher in the IR group than in the non-IR group （P<0.001）. After adjusting for potential confounders including age， sex， and BMI， IR was independently associated with hypertension， dyslipidemia， and hyperuricemia， with odds ratios （OR） and 95% confidence intervals （CI） of 2.57 （1.61-4.09）， 3.81 （2.36-6.17）， and 2.68 （1.53-4.71）， respectively （all P<0.001）. ConclusionInsulin resistance is significantly associated with multiple cardiovascular metabolic risk factors in the middle-aged population. Monitoring IR may contribute to the early prevention and control of cardiovascular diseases.

In the process of maintaining the steady state of bone tissue, the transcription network and signal pathway of the body play a vital role. These complex regulatory mechanisms need precise coordination to ensure the balance between bone formation and bone absorption. Once this balance is broken, it may lead to pathological changes of bone and cartilage, and then lead to various bone diseases. Therefore, it is of great significance to understand these regulatory mechanisms for the prevention and treatment of bone diseases. In recent years, with the deepening of research, more and more lncRNA has been found to be closely related to bone health. Among them, nuclear paraspeckle assembly transcript 1 (NEAT1), as an extremely abundant RNA molecule in mammalian nuclei, has attracted extensive attention. NEAT1 is mainly transcribed from a specific site in human chromosome 11 by RNA polymerase II (RNaseP), which can form two different subtypes NEAT1_1 and NEAT1_2. These two subtypes are different in intracellular distribution and function, but they participate in many biological processes together. Studies have shown that NEAT1 plays a specific role in the process of cell growth and stress response. For example, it can regulate the development of osteoblasts (OB), osteoclasts (OC) and chondrocytes by balancing the differentiation of bone marrow mesenchymal stem cells (BMSCs), thus maintaining the steady state of bone metabolism. This discovery reveals the important role of NEAT1 in bone development and remodeling. In addition, NEAT1 is closely related to a variety of bone diseases. In patients with bone diseases such as osteoporosis (OP), osteoarthritis (OA) and osteosarcoma (OS), the expression level of NEAT1 is different. These differential expressions may be closely related to the pathogenesis and progression of bone diseases. By regulating the level of NEAT1, it can affect a variety of signal transduction pathways, and then affect the development of bone diseases. For example, some studies show that by regulating the expression level of NEAT1, the activity of osteoclasts can be inhibited, and the proliferation and differentiation of osteoblasts can be promoted, thus improving the symptoms of osteoporosis. It is worth noting that NEAT1 can also be used as a key sensor for the prevention and treatment of bone diseases. When exercising or receiving some natural products, the expression level of NEAT1 will change, thus reflecting the response of bones to external stimuli. This feature makes NEAT1 an important target for studying the prevention and treatment strategies of bone diseases. However, although the role of NEAT1 in bone biology and bone diseases has been initially recognized, its specific mechanism and regulatory relationship are still controversial. For example, the expression level, mode of action and interaction with other molecules of NEAT1 in different bone diseases still need further in-depth study. This paper reviews the role of NEAT1 in maintaining bone and cartilage metabolism, and discusses its expression and function in various bone diseases. By combing the existing research results and controversial points, this paper aims to provide new perspectives and ideas for the prevention and treatment of bone diseases, and provide useful reference and enlightenment for future research.

Objective To investigate the effects of strength training and flexibility training on the strategy of crossing obstacles for the elderly women and the risk of tripping over obstacles.Methods Twenty five elderly women were randomly divided into strength training group(n=13)and flexibility training group(n=12),and received corresponding intervention training for 12 weeks.The kinematics data of obstacle crossing were collected using an infrared three-dimensional(3D)motion capture system before and after training.Results Both strength training and flexibility training could significantly improve the gait speed(P=0.033),stride length(P=0.020)and toe distance(P=0.014)during 25 cm obstacle crossing.The interactive effect of training and time was significant for the crossing height(15 cm:P=0.025;25 cm:P=0.019).The interactive effect of training and time was significant for the margin of stability(MOS)in the internal-external direction during 25 cm obstacle crossing(P<0.05).The minimum MOS in the first single support period(P=0.046)and the MOS at the time when the toe crossed directly above the obstacle(P=0.043)in strength training group were significantly increased.Conclusions Both strength training and flexibility training can improve the spatiotemporal characteristics of the elderly women during obstacle crossing.Compared with flexibility,muscle strength is the most important reason that restricts the crossing height of the elderly women.Strength training can effectively reduce the risk of tripping over obstacles by improving the crossing height and dynamic stability of elderly women.

Objective:To evaluate the application effect of spiral cyclic intervention with information support based on knowledge, attitude/belief, practice (KABP) Model on the quality of life among hospitalized people living with HIV/AIDS (PLWHA), in order to provide a scientific basis for improving quality of life intervention strategies.Methods:This was a random-controlled trial. A total of 50 hospitalized PLWHA admitted to the Infectious Diseases Department of Xi'an Eighth Hospital from October 2021 to November 2023 were randomly divided into an intervention group and a control group by random number table method. The intervention group received a 12-week spiral cyclic intervention with information support, while the control group received routine nursing care. The quality of life data were collected at baseline, week 4, and week 12. Data analysis was conducted using Chi-square test, t-test and generalized estimating equation(GEE). Results:A total of 50 PLWHA were enrolled with 25 cases in each group. The control group comprised 22 males and 3 females aged 17-60 years, while the intervention group included 23 males and 2 females aged 21-60 years. GEE results showed that after 12 weeks of intervention, the intervention group scored significantly higher than the control group in physical health summary and eight dimensions: mental health, health distress, quality of life, cognitive functioning, pain, role functioning, general health perceptions, and health transition. Specifically, the intervention group′s scores were (49.45 ± 1.45), (64.99 ± 3.25), (69.29 ± 4.13), (61.71 ± 2.63), (76.46 ± 3.85), (81.54 ± 3.80), (82.05 ± 5.68), (55.80 ± 3.52), and (64.15 ± 3.28) points, compared to the control group′s (45.39 ± 1.17), (52.75 ± 2.19), (54.08 ± 4.15), (51.03 ± 3.07), (65.14 ± 3.36), (71.41 ± 3.39), (60.84 ± 6.15), (44.57 ± 2.40), and (51.43 ± 3.08) points. All differences were statistically significant (Wald χ2 values were 3.97-9.53, all P<0.05). Conclusions:The spiral cyclic intervention with information support based on the KABP Model effectively improves the quality of life of hospitalized PLWHA, particularly in physical health summary.

Objective:To compare the differences in breastfeeding rates and the incidence of clinical complications in very/extremely low birth weight infants with and without the use of donor milk banks.Methods:Before and after the establishment of the donor milk bank,a total of 279 very/extremely low birth weight infants who were hospitalized in neonatal intensive care unit in a tertiary hospital in Beijing were selected.In the study,136 infants who did not receive donated breast-feeding were included in con-trol group and 143 infants who received donated breast-feeding were included in observation group.The clinical data of mothers and their infants were collected.The mother's information included gestational age,maternal comorbidities,and mode of delivery.Infant information includes gender,weight,gesta-tional age,duration of breastfeeding,total enteral feeding time,hospitalization time and incidence of complications(feeding intolerance,necrotizing enterocolitis,retinopathy of prematurity).Results:The maternal ages were(33.5±4.2)years in the observation group and(32.5±3.9)years in the con-trol group.Cesareans were performed in 95 cases(70.4％)and 81 cases(66.9％),respectively.The gestational ages of preterm infants were(29.2±2.1)weeks and(29.1±2.2)weeks,with birth weights of(1 140.5±247.1)g and(1 169.4±228.6)g,respectively.Newborn boys accounted for 72 cases(50.3％)in the observation group and 63 cases(46.3％)in the control group.No statistically significant differences were found in baseline characteristics between the two groups(all P＞0.05).After the use of donor milk banks,the rate of exclusive breastfeeding in very/low birth weight infants increased from 3.1％to 10.5％(x2=5.778,P=0.016)during hospitalization,the time to full enteral feeding was shortened from 13dto10d(Z=-4.567,P＜0.001),the first breastfeeding time was shortened from the third day of admission to the first day of admission(Z=-11.812,P＜0.001),the first breastfeeding of mother's own milk was extended from the third day of admission to the fourth day of admission(Z=-4.652,P＜0.001),and the incidence of feeding intolerance during hospitalization was reduced from 34.0％to 10.0％(x2=17.015,P＜0.001).There were no significant differences in the incidence of necrotizing enterocolitis,late-onset sepsis,retinopathy of prematurity and total length of hospital stay(P＞0.05).Conclusion:The use of donor milk bank can improve the breastfeeding rate,shorten the time to first breastfeeding,and reduce the incidence of feeding intolerance in very/extremely low birth weight infants,which provides a reference for the clinical treatment of very/extremely low birth weight infants.

Objective:To explore the cognitive characteristics, influencing factors, and treatment needs regarding biologic agents among patients with psoriasis.Methods:A cross-sectional study was conducted. Patients with psoriasis attending the Department of Dermatology, Xijing Hospital, Air Force Medical University were selected from October to December 2022, and from June to December 2023. A self-designed electronic questionnaire was used for investigation, covering demographic characteristics, psoriasis history (disease types, disease duration, previous treatments, etc.), biologics knowledge (sources of awareness, core cognitive dimensions), and treatment needs.Results:The valid questionnaire response rate reached 93.2% (439/471). The ages of enrolled patients were 35.95 ± 12.57 years, and the disease duration was 7.90 ± 3.26 years. Psoriasis vulgaris was the predominant type (363 cases, 82.69%). The overall awareness rate of biologics slightly increased from 68.62% (105/153) in 2022 to 72.38% (207/286) in 2023 ( P > 0.05). Primary information sources included new media (WeChat/internet) platforms (168 cases, 53.84%) and peer-to-peer sharing (115 cases, 36.86%), while physician counseling merely accounted for 9.29% (29 cases) ( P < 0.001). Insufficient knowledge of biologics was manifested primarily as poor awareness of comorbidities (47.60%, 209/439) and treatment monitoring protocols (22.32%, 98/439). Core concerns regarding biologic therapy included safety (73.34%, 322/439), economic burden (65.14%, 286/439), and long-term efficacy (63.55%, 279/439) ; 60.13% (264/439) of the patients expected rapid improvement of skin symptoms. As for treatment modalities, 90.20% (396/439) of the patients preferred regimens with extended dosing intervals. Conclusions:The patients with psoriasis demonstrated an imbalance in their cognitive structure regarding biologic agents. Their treatment needs exhibited multidimensional characteristics, emphasizing not only rapid clearance of skin lesions but also greater importance of treatment safety and cost-effectiveness.

A 34-year-old male patient was admitted to the hospital on October 6, 2023 due to fall from a high place. Physical examination showed blood pressure 96/53 mmHg (1 mmHg=0.133 kPa), heart rate of 105 beats/min, slight bulge in the right kidney area, and positive percussion pain in the right kidney area. CT examination showed complete rupture of the lower pole of the right kidney, thrombosis of the right renal vein, and contusion and laceration of the right posterior lobe of the liver. According to the American Association for the Surgery of Trauma renal injury classification, he was diagnosed as grade Ⅳ renal rupture. After blood transfusion and fluid infusion, vital signs were stable. Laparoscopic nephrectomy plus renal vein thrombectomy plus liver rupture repair were performed. The operation took 90 minutes, and about 300 ml of blood clots and fresh blood were aspirated during the operation. The patient's vital signs were stable after the operation, and he could get out of bed 3 days after the operation. CT reexamination 1 month after the operation showed right kidney resected and the liver healed well. This operation is feasible if the patient's vital signs are stable, with the advantages of small surgical incision, short operation time, less bleeding, fast postoperative recovery and good surgical effect.