1.Brain functional changes following electroacupuncture in a mouse model of comorbid pain and depression: A resting-state functional magnetic resonance imaging study.
Xuan YIN ; Xiao-Ling ZENG ; Jing-Jing LIN ; Wen-Qing XU ; Kai-Yu CUI ; Xiu-Tian GUO ; Wei LI ; Shi-Fen XU
Journal of Integrative Medicine 2025;23(2):159-168
OBJECTIVE:
Comorbid pain and depression are common but remain difficult to treat. Electroacupuncture (EA) can effectively improve symptoms of depression and relieve pain, but its neural mechanism remains unclear. Therefore, we used resting-state functional magnetic resonance imaging (rs-fMRI) to detect cerebral changes after initiating a mouse pain model via constriction of the infraorbital nerve (CION) and then treating these animals with EA.
METHODS:
Forty male C57BL/6J mice were divided into 4 groups: control, CION model, EA, and sham acupuncture (without needle insertion). EA was performed on the acupoints Baihui (GV20) and Zusanli (ST36) for 20 min, once a day for 10 consecutive days. The mechanical withdrawal threshold was tested 3 days after the surgery and every 3 days after the intervention. The depressive behavior was evaluated with the tail suspension test, open-field test, elevated plus maze (EPM), sucrose preference test, and marble burying test. The rs-fMRI was used to detect the cerebral changes of the functional connectivity (FC) in the mice following EA treatment.
RESULTS:
Compared with the CION group, the mechanical withdrawal threshold increased in the EA group at the end of the intervention (P < 0.05); the immobility time in tail suspension test decreased (P < 0.05); and the times of the open arm entry and the open arm time in the EPM increased (both P < 0.001). There was no difference in the sucrose preference or marble burying tests (both P > 0.05). The fMRI results showed that EA treatment downregulated the amplitude of low-frequency fluctuations and regional homogeneity values, while these indicators were elevated in brain regions including the amygdala, hippocampus and cerebral cortex in the CION model for comorbid pain and depression. Selecting the amygdala as the seed region, we found that the FC was higher in the CION group than in the control group. Meanwhile, EA treatment was able to decrease the FC between the amygdala and other brain regions including the caudate putamen, thalamus, and parts of the cerebral cortex.
CONCLUSION
EA can downregulate the abnormal activation of neurons in the amygdala and improve its FC with other brain regions, thus exerting analgesic and antidepressant effects. Please cite this article as: Yin X, Zeng XL, Lin JJ, Xu WQ, Cui KY, Guo XT, Li W, Xu SF. Brain functional changes following electroacupuncture in a mouse model of comorbid pain and depression: a resting-state functional magnetic resonance imaging study. J Integr Med. 2025; 23(2): 159-168.
Animals
;
Electroacupuncture
;
Male
;
Magnetic Resonance Imaging
;
Depression/diagnostic imaging*
;
Mice, Inbred C57BL
;
Brain/diagnostic imaging*
;
Disease Models, Animal
;
Mice
;
Pain/diagnostic imaging*
;
Acupuncture Points
2.Association of Co-Exposure to Polycyclic Aromatic Hydrocarbons and Metal(loid)s with the Risk of Neural Tube Defects: A Case-Control Study in Northern China.
Xiao Qian JIA ; Yuan LI ; Lei JIN ; Lai Lai YAN ; Ya Li ZHANG ; Ju Fen LIU ; Le ZHANG ; Linlin WANG ; Ai Guo REN ; Zhi Wen LI
Biomedical and Environmental Sciences 2025;38(2):154-166
OBJECTIVE:
Exposure to polycyclic aromatic hydrocarbons (PAHs) or metal(loid)s individually has been associated with neural tube defects (NTDs). However, the impacts of PAH and metal(loid) co-exposure and potential interaction effects on NTD risk remain unclear. We conducted a case-control study in China among population with a high prevalence of NTDs to investigate the combined effects of PAH and metal(loid) exposures on the risk of NTD.
METHODS:
Cases included 80 women who gave birth to offspring with NTDs, whereas controls were 50 women who delivered infants with no congenital malformations. We analyzed the levels of placental PAHs using gas chromatography and mass spectrometry, PAH-DNA adducts with 32P-post-labeling method, and metal(loid)s with an inductively coupled plasma mass spectrometer. Unconditional logistic regression was employed to estimate the associations between individual exposures and NTDs. Least absolute shrinkage and selection operator (LASSO) penalized regression models were used to select a subset of exposures, while additive interaction models were used to identify interaction effects.
RESULTS:
In the single-exposure models, we found that eight PAHs, PAH-DNA adducts, and 28 metal(loid)s were associated with NTDs. Pyrene, selenium, molybdenum, cadmium, uranium, and rubidium were selected through LASSO regression and were statistically associated with NTDs in the multiple-exposure models. Women with high levels of pyrene and molybdenum or pyrene and selenium exhibited significantly increased risk of having offspring with NTDs, indicating that these combinations may have synergistic effects on the risk of NTDs.
CONCLUSION
Our findings suggest that individual PAHs and metal(loid)s, as well as their interactions, may be associated with the risk of NTDs, which warrants further investigation.
Humans
;
Neural Tube Defects/chemically induced*
;
Polycyclic Aromatic Hydrocarbons/adverse effects*
;
Female
;
Case-Control Studies
;
China/epidemiology*
;
Adult
;
Pregnancy
;
Environmental Pollutants
;
Maternal Exposure/adverse effects*
;
Metals/toxicity*
;
Young Adult
;
Risk Factors
3.Astragaloside IV Alleviates Podocyte Injury in Diabetic Nephropathy through Regulating IRE-1α/NF-κ B/NLRP3 Pathway.
Da-Lin SUN ; Zi-Yi GUO ; Wen-Yuan LIU ; Lin ZHANG ; Zi-Yuan ZHANG ; Ya-Ling HU ; Su-Fen LI ; Ming-Yu ZHANG ; Guang ZHANG ; Jin-Jing WANG ; Jing-Ai FANG
Chinese journal of integrative medicine 2025;31(5):422-433
OBJECTIVE:
To investigate the effects of astragaloside IV (AS-IV) on podocyte injury of diabetic nephropathy (DN) and reveal its potential mechanism.
METHODS:
In in vitro experiment, podocytes were divided into 4 groups, normal, high glucose (HG), inositol-requiring enzyme 1 (IRE-1) α activator (HG+thapsigargin 1 µmol/L), and IRE-1α inhibitor (HG+STF-083010, 20 µmol/L) groups. Additionally, podocytes were divided into 4 groups, including normal, HG, AS-IV (HG+AS-IV 20 µmol/L), and IRE-1α inhibitor (HG+STF-083010, 20 µmol/L) groups, respectively. After 24 h treatment, the morphology of podocytes and endoplasmic reticulum (ER) was observed by electron microscopy. The expressions of glucose-regulated protein 78 (GRP78) and IRE-1α were detected by cellular immunofluorescence. In in vivo experiment, DN rat model was established via a consecutive 3-day intraperitoneal streptozotocin (STZ) injections. A total of 40 rats were assigned into the normal, DN, AS-IV [AS-IV 40 mg/(kg·d)], and IRE-1α inhibitor [STF-083010, 10 mg/(kg·d)] groups (n=10), respectively. The general condition, 24-h urine volume, random blood glucose, urinary protein excretion rate (UAER), urea nitrogen (BUN), and serum creatinine (SCr) levels of rats were measured after 8 weeks of intervention. Pathological changes in the renal tissue were observed by hematoxylin and eosin (HE) staining. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to detect the expressions of GRP78, IRE-1α, nuclear factor kappa Bp65 (NF-κBp65), interleukin (IL)-1β, NLR family pyrin domain containing 3 (NLRP3), caspase-1, gasdermin D-N (GSDMD-N), and nephrin at the mRNA and protein levels in vivo and in vitro, respectively.
RESULTS:
Cytoplasmic vacuolation and ER swelling were observed in the HG and IRE-1α activator groups. Podocyte morphology and ER expansion were improved in AS-IV and IRE-1α inhibitor groups compared with HG group. Cellular immunofluorescence showed that compared with the normal group, the fluorescence intensity of GRP78 and IRE-1α in the HG and IRE-1α activator groups were significantly increased whereas decreased in AS-IV and IRE-1α inhibitor groups (P<0.05). Compared with the normal group, the mRNA and protein expressions of GRP78, IRE-1α, NF-κ Bp65, IL-1β, NLRP3, caspase-1 and GSDMD-N in the HG group was increased (P<0.05). Compared with HG group, the expression of above indices was decreased in the AS-IV and IRE-1α inhibitor groups, and the expression in the IRE-1α activator group was increased (P<0.05). The expression of nephrin was decreased in the HG group, and increased in AS-IV and IRE-1α inhibitor groups (P<0.05). The in vivo experiment results revealed that compared to the normal group, the levels of blood glucose, triglyceride, total cholesterol, BUN, blood creatinine and urinary protein in the DN group were higher (P<0.05). Compared with DN group, the above indices in AS-IV and IRE-1α inhibitor groups were decreased (P<0.05). HE staining revealed glomerular hypertrophy, mesangial widening and mesangial cell proliferation in the renal tissue of the DN group. Compared with the DN group, the above pathological changes in renal tissue of AS-IV and IRE-1α inhibitor groups were alleviated. Quantitative RT-PCR and Western blot results of GRP78, IRE-1α, NF-κ Bp65, IL-1β, NLRP3, caspase-1 and GSDMD-N were consistent with immunofluorescence analysis.
CONCLUSION
AS-IV could reduce ERS and inflammation, improve podocyte pyroptosis, thus exerting a podocyte-protective effect in DN, through regulating IRE-1α/NF-κ B/NLRP3 signaling pathway.
Podocytes/metabolism*
;
Animals
;
Diabetic Nephropathies/metabolism*
;
Saponins/therapeutic use*
;
Triterpenes/therapeutic use*
;
Signal Transduction/drug effects*
;
NF-kappa B/metabolism*
;
Protein Serine-Threonine Kinases/metabolism*
;
Male
;
Rats, Sprague-Dawley
;
NLR Family, Pyrin Domain-Containing 3 Protein/metabolism*
;
Endoribonucleases/metabolism*
;
Endoplasmic Reticulum Chaperone BiP
;
Rats
;
Diabetes Mellitus, Experimental/complications*
;
Endoplasmic Reticulum/metabolism*
;
Multienzyme Complexes
4.The Mesencephalic Locomotor Region for Locomotion Control
Xing-Chen GUO ; Yan XIE ; Xin-Shuo WEI ; Wen-Fen LI ; Ying-Yu SUN
Progress in Biochemistry and Biophysics 2025;52(7):1804-1816
Locomotion, a fundamental motor function encompassing various forms such as swimming, walking, running, and flying, is essential for animal survival and adaptation. The mesencephalic locomotor region (MLR), located at the midbrain-hindbrain junction, is a conserved brain area critical for controlling locomotion. This review highlights recent advances in understanding the MLR’s structure and function across species, from lampreys to mammals and birds, with a particular focus on insights gained from optogenetic studies in mammals. The goal is to uncover universal strategies for MLR-mediated locomotor control. Electrical stimulation of the MLR in species such as lampreys, salamanders, cats, and mice initiates locomotion and modulates speed and patterns. For example, in lampreys, MLR stimulation induces swimming, with increased intensity or frequency enhancing propulsive force. Similarly, in salamanders, graded stimulation transitions locomotor outputs from walking to swimming. Histochemical studies reveal that effective MLR stimulation sites colocalize with cholinergic neurons, suggesting a conserved neurochemical basis for locomotion control. In mammals, the MLR comprises two key nuclei: the cuneiform nucleus (CnF) and the pedunculopontine nucleus (PPN). Both nuclei contain glutamatergic and GABAergic neurons, with the PPN additionally housing cholinergic neurons. Optogenetic studies in mice by selectively activating glutamatergic neurons have demonstrated that the CnF and PPN play distinct roles in motor control: the CnF drives rapid escape behaviors, while the PPN regulates slower, exploratory movements. This functional specialization within the MLR allows animals to adapt their locomotion patterns and speed in response to environmental demands and behavioral objectives. Similar to findings in lampreys, the CnF and PPN in mice transmit motor commands to spinal effector circuits by modulating the activity of brainstem reticular formation neurons. However, they achieve this through distinct reticulospinal pathways, enabling the generation of specific behaviors. Further insights from monosynaptic rabies viral tracing reveal that the CnF and PPN integrate inputs from diverse brain regions to produce context-appropriate behaviors. For instance, glutamatergic neurons in the PPN receive signals from other midbrain structures, the basal ganglia, and medullary nuclei, whereas glutamatergic neurons in the CnF rarely receive inputs from the basal ganglia but instead are strongly influenced by the periaqueductal grey and inferior colliculus within the midbrain. These differential connectivity patterns underscore the specialized roles of the CnF and PPN in motor control, highlighting their unique contributions to coordinating locomotion. Birds exhibit exceptional flight capabilities, yet the avian MLR remains poorly understood. Comparative studies suggest that the pedunculopontine tegmental nucleus (PPTg) in birds is homologous to the mammalian PPN, which contains cholinergic neurons, while the intercollicular nucleus (ICo) or nucleus isthmi pars magnocellularis (ImC) may correspond to the CnF. These findings provide important clues for identifying the avian MLR and elucidating its role in flight control. However, functional validation through targeted experiments is urgently needed to confirm these hypotheses. Optogenetics and other advanced techniques in mice have greatly advanced MLR research, enabling precise manipulation of specific neuronal populations. Future studies should extend these methods to other species, particularly birds, to explore unique locomotor adaptations. Comparative analyses of MLR structure and function across species will deepen our understanding of the conserved and evolved features of motor control, revealing fundamental principles of locomotion regulation throughout evolution. By integrating findings from diverse species, we can uncover how the MLR has been adapted to meet the locomotor demands of different environments, from aquatic to aerial habitats.
5.Construction and validation of a risk prediction model for in-hospital death after successful resuscitation in patients with cardiac arrest
Yu LI ; Zhen CHEN ; Xin GUO ; Yifan LIANG ; Jueyan WANG ; Jinlei LI ; Xianting YANG ; Fen AI
Journal of Clinical Medicine in Practice 2025;29(11):26-32,41
Objective To construct and validate a risk prediction model for in-hospital death af-ter successful resuscitation in patients with cardiac arrest.Methods A retrospective study was con-ducted on 295 patients with cardiac arrest who successfully restored spontaneous circulation after car-diopulmonary resuscitation and were further treated in hospital.The patients were divided into training and validation sets using K-fold cross-validation and then grouped and compared based on whether in-hospital death occurred.A binary Logistic regression analysis was used to screen risk prediction fac-tors,and a nomogram prediction model was constructed.The model performance was evaluated and validated in the training and validation sets,respectively.Results The results of the multivariate Logistic regression analysis showed that hospitalization duration(OR=1.180;95%CI,1.080 to 1.280;P<0.001),norepinephrine dose(OR=0.980;95%CI,0.970 to 0.990;P=0.002),ini-tial respiratory rate after resuscitation(OR=1.090;95%CI,1.030 to 1.150;P=0.004),and sinus rhythm recovery after resuscitation(OR=4.280;95%CI,1.670 to 10.980;P=0.003)were inde-pendent influencing factors for in-hospital death.A nomogram model was constructed based on these in-dependent influencing factors,and it was verified that the model had good discrimination,calibration,applicability,and rationality.Conclusion The influencing factors for in-hospital death after successful resuscitation in patients with cardiac arrest include hospitalization duration,norepinephrine dose,initial respiratory rate after resuscitation,and sinus rhythm recovery after resuscitation.The nomo-gram model constructed based on these factors can provide a reference for clinical decision-making.
6.Quantitative evaluation of left ventricular remodeling in left bundle branch block induced cardiomyopathy by multimodal imaging
Xiaoxian WANG ; Changqing MIAO ; Beibei GE ; Mingxia LI ; Fen CHEN ; Fang XU ; Ning ZHANG ; Guanjun GUO ; Wei XU ; Wenzhi SHEN ; Yingming ZHAO ; Jing YAO
Chinese Journal of Ultrasonography 2025;34(1):26-32
Objective:To quantitatively evaluate the effects of left bundle branch block(LBBB)on left ventricular structure,function and myocardial perfusion using left ventricular pressure-strain loop and single photon emission computed tomography(SPECT),and to investigate the relationship between myocardial work,myocardial perfusion and pathological changes of left ventricular remodeling in left bundle branch block induced cardiomyopathy(LBBB-CM).Methods:Fourteen male beagle dogs were selected,and the main trunk of the left bundle branch was ablated to create an LBBB dog model. Electrocardiogram(ECG),transesophageal echocardiography and arterial blood pressure data of LBBB dogs were collected before and 12 months after the ablation of left bundle branch trunk. Global and segmental myocardial work parameters were obtained by left ventricular pressure-strain loop. The differences of above parameters between baseline and 12 months after the ablation of left bundle branch were compared. SPECT was performed in LBBB dogs 12 months after the creation of LBBB. The hearts were harvested for anatomy observation and histopathological analysis in LBBB dogs and another 7 male beagle dogs(normal control group)matched by age and weight. The correlation between myocardial perfusion(percentage of regional tracer uptake)and myocardial work parameters,myocardial fibrosis in LBBB dogs were analyzed.Results:Compared with baseline,the left ventricular end-diastolic volume of 12 months after the ablation increased[(20.78 ± 5.32)ml vs(26.71 ± 7.94)ml, P = 0.003],left ventricular ejection fraction decreased[(59.17 ± 5.67)% vs(47.69 ± 5.45)%, P<0.001];left ventricular global/segmental longitudinal strain,global/segmental constructive work and global/segmental work efficiency decreased(all P<0.05),left ventricular global/segmental wasted work increased(all P<0.001). Heterogenous perfusion defect was observed in LBBB dogs by SPECT,compared with lateral wall segments,the percentage of regional tracer uptake of septum was decreased(all P<0.05). Gross anatomical and myocardial pathological changes were manifested as cardiomegaly,flaky or focal grayish thickening of endocardium,cardiomyocyte degeneration and fibrosis. Compared with normal control group,the collagen fiber volume fraction(CVF)in all segmental endocardium and partial segmental myocardium of LBBB dogs were significantly increased(all P<0.05). Percentage of regional tracer uptake was positively correlated with segmental myocardial work(SMW)and segmental myocardial efficiency(SWE)( r s = 0.49,0.31;both P<0.001),and negatively correlated with CVF and segmental wasted work(SWW)( r s = -0.51,-0.49;both P<0.001). Conclusions:Isolated LBBB is not benign,which can result in left ventricular remodeling,decreased cardiac constructive function,abnormal myocardial perfusion,endocardial fibrosis and myocardial fibrosis.The parameters of myocardial work assecsed by echocardiograpgy and myocardial perfusion,as non-invasive examination,can to some extent reflect the degree of left ventricular remodeling in LBBB-CM.
7.Quantitative evaluation of left ventricular remodeling in left bundle branch block induced cardiomyopathy by multimodal imaging
Xiaoxian WANG ; Changqing MIAO ; Beibei GE ; Mingxia LI ; Fen CHEN ; Fang XU ; Ning ZHANG ; Guanjun GUO ; Wei XU ; Wenzhi SHEN ; Yingming ZHAO ; Jing YAO
Chinese Journal of Ultrasonography 2025;34(1):26-32
Objective:To quantitatively evaluate the effects of left bundle branch block(LBBB)on left ventricular structure,function and myocardial perfusion using left ventricular pressure-strain loop and single photon emission computed tomography(SPECT),and to investigate the relationship between myocardial work,myocardial perfusion and pathological changes of left ventricular remodeling in left bundle branch block induced cardiomyopathy(LBBB-CM).Methods:Fourteen male beagle dogs were selected,and the main trunk of the left bundle branch was ablated to create an LBBB dog model. Electrocardiogram(ECG),transesophageal echocardiography and arterial blood pressure data of LBBB dogs were collected before and 12 months after the ablation of left bundle branch trunk. Global and segmental myocardial work parameters were obtained by left ventricular pressure-strain loop. The differences of above parameters between baseline and 12 months after the ablation of left bundle branch were compared. SPECT was performed in LBBB dogs 12 months after the creation of LBBB. The hearts were harvested for anatomy observation and histopathological analysis in LBBB dogs and another 7 male beagle dogs(normal control group)matched by age and weight. The correlation between myocardial perfusion(percentage of regional tracer uptake)and myocardial work parameters,myocardial fibrosis in LBBB dogs were analyzed.Results:Compared with baseline,the left ventricular end-diastolic volume of 12 months after the ablation increased[(20.78 ± 5.32)ml vs(26.71 ± 7.94)ml, P = 0.003],left ventricular ejection fraction decreased[(59.17 ± 5.67)% vs(47.69 ± 5.45)%, P<0.001];left ventricular global/segmental longitudinal strain,global/segmental constructive work and global/segmental work efficiency decreased(all P<0.05),left ventricular global/segmental wasted work increased(all P<0.001). Heterogenous perfusion defect was observed in LBBB dogs by SPECT,compared with lateral wall segments,the percentage of regional tracer uptake of septum was decreased(all P<0.05). Gross anatomical and myocardial pathological changes were manifested as cardiomegaly,flaky or focal grayish thickening of endocardium,cardiomyocyte degeneration and fibrosis. Compared with normal control group,the collagen fiber volume fraction(CVF)in all segmental endocardium and partial segmental myocardium of LBBB dogs were significantly increased(all P<0.05). Percentage of regional tracer uptake was positively correlated with segmental myocardial work(SMW)and segmental myocardial efficiency(SWE)( r s = 0.49,0.31;both P<0.001),and negatively correlated with CVF and segmental wasted work(SWW)( r s = -0.51,-0.49;both P<0.001). Conclusions:Isolated LBBB is not benign,which can result in left ventricular remodeling,decreased cardiac constructive function,abnormal myocardial perfusion,endocardial fibrosis and myocardial fibrosis.The parameters of myocardial work assecsed by echocardiograpgy and myocardial perfusion,as non-invasive examination,can to some extent reflect the degree of left ventricular remodeling in LBBB-CM.
8.Analysis of urinary iodine level in Hashimoto thyroiditis patients
Xiaodie Li ; Yongxia Xu ; Fen Wang ; Wenlu Guo ; Wei Jia ; Xuefeng Wang ; Lang Lang ; Defa Zhu
Acta Universitatis Medicinalis Anhui 2024;59(1):144-148
Objective :
To analyze the difference of urinary iodine level in Hashimoto thyroiditis ( HT) patients, and to explore the possible relationship between urinary iodine level and HT under different iodine nutritional sta- tus,so as to provide some references for reasonable iodine intake in HT patients.
Methods :
A total of 101 hospi- talized HT patients were selected as HT group and divided into 3 groups according to thyroid function : HT group with hyperthyroidism (41 cases) .There were 25 cases in HT group with normal thyroid function.There were 35 cases in HT combined with hypothyroidism group.In addition,30 healthy subjects were selected as control group. Serum levels of thyroid stimulating hormone(TSH) ,triiodothyronine(T3 ) ,thyroxine (T4 ) ,thyroid peroxidase an- tibody (TPOAb) and thyroglobulin antibody (ATG) were detected by chemiluminescence assay.The size and mor- phological structure of thyroid organs were examined by ultrasonography.Urinary iodine was determined by catalytic spectrophotometry with arsenic and cerium.The nutritional status of iodine was classified into iodine deficiency ( < 100 μg/ L) ,iodine adequacy( 100 -199 μg/ L) ,iodine adequacy (200 -299 μg/ L) and iodine excess ( ≥ 300 μg/ L) .Non-parametric test was used to compare urinary iodine level between HT group and control group,one- way ANOVA and t test were used to compare urinary iodine level between HT group and control group ,and Spearman correlation analysis was used to compare the correlation between urinary iodine level and T3 ,T4 ,TSH, ATG and TPOAb under different iodine nutrition status.
Results :
Compared with control group,ATG and TPOAb levels in HT group increased (P<0. 001) ,and urinary iodine levels increased (P<0. 05) ,with statistical signifi- cance.Compared with the control group in different thyroid function states,only the HT group with hypothyroidism increased the urinary iodine level (P<0. 01) ,and the difference was statistically significant.Spearman correlation analysis showed that urine iodine level was positively correlated with ATG and TPOAb levels in iodine excess condi- tion (P<0. 05) ,and urine iodine level was positively correlated with TSH level in iodine sufficient condition and iodine excess condition in HT patients (P<0. 05) .
Conclusion
The urinary iodine level of HT patients was high- er than that of normal people.When the urinary iodine level of residents is ≥ 300 μg/ L,iodine intake is prone to HT.When the urinary iodine level of HT patients is ≥ 200 μg/ L,iodine consumption is prone to hypothyroidism, and iodine intake should be limited.
9.Stability study of umbilical cord mesenchymal stem cells formulation in large-scale production
Wang-long CHU ; Tong-jing LI ; Yan SHANGGUAN ; Fang-tao HE ; Jian-fu WU ; Xiu-ping ZENG ; Tao GUO ; Qing-fang WANG ; Fen ZHANG ; Zhen-zhong ZHONG ; Xiao LIANG ; Jun-yuan HU ; Mu-yun LIU
Acta Pharmaceutica Sinica 2024;59(3):743-750
Umbilical cord mesenchymal stem cells (UC-MSCs) have been widely used in regenerative medicine, but there is limited research on the stability of UC-MSCs formulation during production. This study aims to assess the stability of the cell stock solution and intermediate product throughout the production process, as well as the final product following reconstitution, in order to offer guidance for the manufacturing process and serve as a reference for formulation reconstitution methods. Three batches of cell formulation were produced and stored under low temperature (2-8 ℃) and room temperature (20-26 ℃) during cell stock solution and intermediate product stages. The storage time intervals for cell stock solution were 0, 2, 4, and 6 h, while for intermediate products, the intervals were 0, 1, 2, and 3 h. The evaluation items included visual inspection, viable cell concentration, cell viability, cell surface markers, lymphocyte proliferation inhibition rate, and sterility. Additionally, dilution and culture stability studies were performed after reconstitution of the cell product. The reconstitution diluents included 0.9% sodium chloride injection, 0.9% sodium chloride injection + 1% human serum albumin, and 0.9% sodium chloride injection + 2% human serum albumin, with dilution ratios of 10-fold and 40-fold. The storage time intervals after dilution were 0, 1, 2, 3, and 4 h. The reconstitution culture media included DMEM medium, DMEM + 2% platelet lysate, 0.9% sodium chloride injection, and 0.9% sodium chloride injection + 1% human serum albumin, and the culture duration was 24 h. The evaluation items were viable cell concentration and cell viability. The results showed that the cell stock solution remained stable for up to 6 h under both low temperature (2-8 ℃) and room temperature (20-26 ℃) conditions, while the intermediate product remained stable for up to 3 h under the same conditions. After formulation reconstitution, using sodium chloride injection diluted with 1% or 2% human serum albumin maintained a viability of over 80% within 4 h. It was observed that different dilution factors had an impact on cell viability. After formulation reconstitution, cultivation in medium with 2% platelet lysate resulted in a cell viability of over 80% after 24 h. In conclusion, the stability of cell stock solution within 6 h and intermediate product within 3 h meets the requirements. The addition of 1% or 2% human serum albumin in the reconstitution diluent can better protect the post-reconstitution cell viability.
10.Clinical analysis of children with anomalous origin of coronary artery over ten years in a single center
Jieliang LIN ; Meng ZHANG ; Fen LI ; Lijun FU ; Wei GAO ; Tingliang LIU ; Ying GUO ; Yumin ZHONG ; Jie SHEN
Chinese Journal of Applied Clinical Pediatrics 2024;39(10):757-760
Objective:To analyze the clinical characteristics of different types of anomalous origin of the coronary artery.Methods:A case-series study was conducted.Based on the clinical data of children diagnosed with anomalous origin of the coronary artery at Shanghai Children′s Medical Center, Shanghai Jiao Tong University School of Medicine from January 2013 to January 2023, the diseases of different types of anomalous origin of the coronary artery were summarized.Results:A total of 177 children with anomalous origin of the coronary artery were treated.Among them, 122 children developed the anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA), including 54 males and 68 females, with a median age of 1.2 years; 6 children developed the anomalous origin of the right coronary artery from the pulmonary artery (ARCAPA), including 3 males and 3 females, with a median age of 4.3 years; 9 children developed the anomalous left coronary artery from the right coronary sinus (ALCA), including 6 males and 3 females, with a median age of 9.5 years; 40 children developed the anomalous right coronary artery from the left coronary sinus (ARCA), including 24 males and 16 females, with a median age of 7.7 years.Most children diagnosed with ALCAPA had onset within 1 year of age, with chronic heart failure as the main manifestation, and young children were often accompanied by severe mitral regurgitation.A total of 111 children underwent surgery, and 11 children died.Six children with ARCAPA had no obvious clinical symptoms and were treated by operation according to the principle of double coronary circulation after diagnosis.Nine children with ALCA started with syncope, chest pain or abdominal pain after exercise.Eight of the children underwent surgical treatment, including 1 who received a heart transplant.Of the 40 children with ARCA, 23 children had clinical manifestations, with chest tightness, syncope, and chest pain after exercise as chief complaints; 16 children were tested positive for treadmill exercise before surgery; and a total of 13 children received surgical treatment.Conclusions:Different types of anomalous origin of the coronary artery vary in severity.The clinical manifestations of the anomalous origin of the left coronary artery are generally serious, and most of such patients have the risk of cardiac insufficiency or sudden death.Once diagnosed, surgical treatment should be performed timely.The clinical manifestations of the anomalous origin of the right coronary artery are relatively mild, and only a few may have serious consequences, which are usually treated according to the principle of individualization or double coronary circulation.


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