1.Recurrent Diabetic Ketoacidosis: Predictors and Clinical Outcomes in a 24-Year Retrospective Cohort
Liang Wei Wong ; Lisa Mohamed Nor ; Raja Nurazni binti Raja Azwan ; Adilah Zulaikha binti Abd Latib ; Hidayatil Alimi bin Keya Nordin ; Qin Zhi Lee ; Kean Heng Lim ; Jia Ling Low ; Mohd Fyzal bin Bahrudin ; Syaza binti Izhar Hisham ; Jia Whey Jacelyn Ong ; Chin Voon Tong
Journal of the ASEAN Federation of Endocrine Societies 2026;41(S1):33-34
Introduction:
Diabetic ketoacidosis (DKA) is a life-threatening complication associated with significant morbidity and healthcare
burden. Despite advances in diabetes care, recurrent
DKA remains common, often reflecting gaps in treatment
adherence and patient education. Identifying predictors
of recurrence is crucial for risk stratification and targeted
intervention.
Methodology:
We conducted a retrospective observational study of all
adult DKA admissions to a tertiary centre between 2001
and 2025. Electronic medical records were reviewed for
demographic data, biochemical parameters, precipitating
factors, and clinical outcomes. DKA was defined using standard biochemical criteria. Recurrent DKA was defined as ≥2 admissions during the study period. Factors associated
with recurrent DKA admissions were analyzed. Patients
under the age of 18 years and those with missing vital
information were excluded.
Results:
A total of 667 DKA admissions, comprising 566 patients,
were identified, of which 101 admissions (15.1%) were
recurrent, involving 65 patients. Among recurrent DKA
episodes, the most common precipitating factors were
infection (64.4%) and insulin omission (62.4%). After
multivariate analyses, patients with type 1 diabetes
mellitus (T1DM) were more likely to develop recurrent
DKA compared to those with type 2 diabetes mellitus
(aOR 4.16; 95% confidence interval [CI] 2.58–6.70; p <0.001).
Insulin omission was strongly associated with recurrent
DKA (aOR 2.29; 95% CI 1.46–3.60; p <0.001). In contrast,
baseline glycated hemoglobin and chronic kidney disease
were not significantly associated with recurrence. Diabetic
counseling during the first DKA admission did not reduce
recurrent DKA. There were no significant differences in
mortality (3.9% vs 6.2%, p = 0.524) or critical care admission
rates (40.6% vs 38.7%, p = 0.718) between recurrent and first
DKA episodes.
Conclusion
Recurrent DKA accounts for a substantial proportion of
DKA admissions and is strongly associated with insulin
omission and T1DM. Our findings suggest that recurrent
DKA is driven predominantly by behavioral and adherencerelated factors, indicating the need for multidisciplinary
interventions beyond standard inpatient counseling.
Diabetic Ketoacidosis
;
Retrospective Studies
2.Clinical and Biochemical Characteristics of Adult Diabetic Ketoacidosis: T1DM vs. T2DM
Mohd Fyzal Bahrudin ; Chin Voon Tong ; Raja Nurazni Raja Azwan ; Adilah Zulaikha Abd Latib ; idayatil Alimi Keya Nordin ; Qin Zhi Lee ; Kean Heng Lim ; Jia Ling Low ; Syaza Izhar Hisham ; Liang Wei Wong ; Jia Whey Jacelyn Ong ; Lisa Mohamed Nor ; Zanariah Hussein
Journal of the ASEAN Federation of Endocrine Societies 2026;41(S1):39-40
Introduction:
The rising incidence of diabetic ketoacidosis (DKA) in type 2
diabetes mellitus (T2DM) represents a paradigm shift from
its traditional recognition as a hallmark complication of
type 1 diabetes mellitus (T1DM). However, contemporary
data comparing clinical presentation, precipitating factors,
and outcomes between these two populations remain
limited.
Methodology:
In this retrospective observational study, all adult DKA
admissions with T1DM or T2DM at a tertiary centre between
2001 and 2025 were studied. DKA was defined according
to standard biochemical criteria. Electronic medical records were reviewed to extract demographic data, biochemical
parameters, precipitating factors, management details, and
clinical outcomes of all adult DKA admission in T1DM and
T2DM. Patients aged <18 years or those with incomplete
data were excluded.
Results:
A total of 601 DKA episodes were analyzed, comprising
130 (21.6%) in patients with T1DM and 471 (78.4%) in those
with T2DM. Mean age was 26.9 ± 0.71 years for T1DM and
52.1 ± 0.7 years for T2DM. Gender distribution was balanced
(male 48.3%, female 51.7%). Mean hemoglobin A1c (HbA1c)
was 11.3 ± 0.3% in T1DM and 12.1 ± 0.2% in T2DM. Infection
was the most common precipitating factor overall (69.0%),
occurring more frequently in T2DM than in T1DM (74.7%
vs. 56.2%), followed by medication non-adherence (62.4%
vs. 43.8%). Significant differences were observed between
groups in admission pH, bicarbonate (both p <0.001), blood
ketones (p = 0.028), and HbA1c (p = 0.010), whereas anion
gap (p = 0.056) and blood glucose levels (p = 0.755) did not
differ significantly. Clinical outcomes were comparable
with respect to intensive care unit (ICU) admission rates
(40.0% in T1DM vs. 39.0% in T2DM, p = 0.779) and median
resolution time (p = 0.462). However, the median length of
hospital stay was significantly longer in T2DM (8.2 ± 0.32
vs. 5.4 ± 0.4 days; p <0.001). Overall mortality was 6.0%,
with substantially higher mortality in T2DM compared to
T1DM (7.4% vs. 0.8%, p = 0.013).
Conclusion
In this large regional series of adult DKA, most episodes
occurred in T2DM. Despite similar ICU admission rates
and time to resolution, T2DM was associated with more
frequent infection-related precipitants, longer hospital
stays, and higher mortality, underscoring the need for
targeted preventive strategies in this population.
Adult
;
Diabetes Mellitus, Type 1
;
Diabetic Ketoacidosis
;
Diabetes Mellitus, Type 2
3.Diabetic Ketoacidosis in Pregnancy: Clinical Triggers, Outcomes, and Missed Opportunities—A Case Series
Jia Whey Jacelyn Ong ; Chin Voon Tong ; Raja Nurazni binti Raja Azwan ; Adilah Zulaikha binti Abd Latib ; Hidayatil Alimi bin Keya Nordin ; Qin Zhi Lee ; Kean Heng Lim ; Jia Ling Low ; Mohd Fyzal bin Bahrudin ; Syaza binti Izhar Hisham ; Liang Wei Wong ; Lisa Mohamed Nor ; Nurain Mohd Noorr
Journal of the ASEAN Federation of Endocrine Societies 2026;41(S1):48-49
Introduction:
Diabetic ketoacidosis (DKA) in pregnancy is an uncommon
yet life-threatening emergency, with disproportionate risks
to both mother and fetus. Pregnancy-specific physiological changes predispose patients to rapid metabolic decompensation, often with atypical presentations. Despite
this, local data remain limited. We describe the clinical
profile, precipitating factors, and outcomes of DKA in
pregnancy in a tertiary centre, with emphasis on potentially
preventable triggers.
Cases:
Nine pregnant patients with DKA were identified from a
retrospective review of all cases admitted for DKA from
2002 to 2025. Mean age was 31.67 ± 5.20 years; all were
Malay. The majority had type 2 diabetes mellitus (55.6%),
followed by type 1 diabetes (33.3%) and latent autoimmune
diabetes in adults (11.1%). The mean period of amenorrhea
was 19.67 ± 12.62 weeks.
Infection was the leading precipitant (44.4%), with
additional triggers including insulin omission (22.2%),
hyperemesis gravidarum, preterm labor, steroid exposure,
and perioperative fasting. Most diagnoses were made in
the emergency department (55.6%).
Biochemical parameters reflected significant severity (mean
bicarbonate 7.89 ± 2.98 mmol/L; anion gap 25.00 ± 5.81),
with 88.9% classified as severe DKA. Intensive Care Unit
(ICU) care was required in 77.8% of cases. The majority
(77.8%) were admitted to the ICU unit, with a median time
to resolution of 13.00 ± 12.00 hours (interquartile range
[IQR]), and the median hospital length of stay was 7.00 ±
5.00 days (IQR).
Complications during treatment included hypokalemia
(33.3%), acute kidney injury (22.2%), and hypoglycemia
(11.1%). Rebound DKA occurred in one-third of patients.
All patients were discharged clinically stable. Outcome
data demonstrated pregnancy loss in three cases and one
preterm birth.
Conclusion
DKA in pregnancy remains a severe and resource-intensive
condition. This series highlights missed opportunities in
prevention, with modifiable precipitants such as infection
and insulin omission commonly identified. The high
severity at presentation suggests delays in recognition.
Early detection, optimized metabolic care, and targeted
preventive strategies are crucial to improving maternal
and fetal outcomes.
Female
;
Pregnancy
;
Diabetic Ketoacidosis
4.Effect of Slicing Angle and Initial Water Content on Water Migration and Effective Ingredient Content in Drying Process of Salviae Miltiorrhizae Radix et Rhizoma
Guohong YANG ; Bingqian ZHOU ; Heng LU ; Xiao WANG ; Lanping GUO ; Wei LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):208-216
ObjectiveTo explore the effects of angle and original moisture content on the moisture distribution, migration and contents of effective components in the drying process of sliced Salviae Miltiorrhizae Radix et Rhizoma(SMRR). MethodsSet the slicing angles of SMRR at 30°, 45°, and 90°. Cut the fresh samples, 1/3 dehydrated samples, and 2/3 dehydrated samples, dry them in an oven at 40 ℃ and take samples at the set time points. Low-field nuclear magnetic resonance(LF-NMR) and magnetic resonance imaging(MRI) were used to analyze the changes in transverse relaxation time(T2) of SMRR samples in 9 treatment groups at specific times, as well as the distribution and migration of water in the samples. The contents of tanshinone ⅡA, tanshinone Ⅰ, cryptotanshinone, and salvianolic acid B in samples from 9 different treatment groups were determined by high performance liquid chromatography(HPLC), and the best processing technology of SMRR was screened by combining with One-way ANOVA, Duncan multiple comparison and principal component analysis(PCA). ResultsThe moisture content of dry basis of SMRR in each treatment group decreased with the extension of drying time. The drying rate of fresh cut group decreased slowly at first, while the drying rate of water loss group showed a trend of increasing at first and then decreasing. The internal water of SMRR could be divided into three states, including bound water, non flowing water and free water. During the drying process, the water migration law showed that the free water of fresh cut group disappeared after drying for 12 h, the content of bound water gradually decreased, and the overall fluidity deteriorated. In the water loss group, part of the free water was transformed into more cohesive and non flowing water after drying for 3 h, and the three kinds of water basically disappeared after drying for 12 h. The MRI results showed that the entire dehydration process slowly moved from the outer side to the center, and the internal water eventually dissipated. In terms of the contents of active ingredients, the order of the effect of slicing angle on the total content of active ingredients in SMRR was 30°>45°>90°. The content of tanshinones was ranked as 1/3 dehydrated group>2/3 dehydrated group>fresh cut group, and the content of salvianolic acid B was ranked as 1/3 dehydrated group>fresh cut group>2/3 dehydrated group. Combined with the results of PCA and comprehensive scoring results, the overall level of effective component content in SMRR was the highest when cut at 30° after 1/3 of water loss. ConclusionAfter comprehensive evaluation, SMRR can be sliced at 30° after 1/3 of water loss. It is not only easy to cut, but also the surface and cross-sectional colors remain basically unchanged after drying, which is similar to the color under traditional processing, and the effective ingredients are preserved the highest. This study can provide a basis for the optimization of processing technology of SMRR.
5.Beneficial Effects of Dendrobium officinale Extract on Insomnia Rats Induced by Strong Light and Noise via Regulating GABA and GABAA Receptors.
Heng-Pu ZHOU ; Jie SU ; Ke-Jian WEI ; Su-Xiang WU ; Jing-Jing YU ; Yi-Kang YU ; Zhuang-Wei NIU ; Xiao-Hu JIN ; Mei-Qiu YAN ; Su-Hong CHEN ; Gui-Yuan LYU
Chinese journal of integrative medicine 2025;31(6):490-498
OBJECTIVE:
To explore the therapeutic effects and underlying mechanisms of Dendrobium officinale (Tiepi Shihu) extract (DOE) on insomnia.
METHODS:
Forty-two male Sprague-Dawley rats were randomly divided into 6 groups (n=7 per group): normal control, model control, melatonin (MT, 40 mg/kg), and 3-dose DOE (0.25, 0.50, and 1.00 g/kg) groups. Rats were raised in a strong-light (10,000 LUX) and -noise (>80 db) environment (12 h/d) for 16 weeks to induce insomnia, and from week 10 to week 16, MT and DOE were correspondingly administered to rats. The behavior tests including sodium pentobarbital-induced sleep experiment, sucrose preference test, and autonomous activity test were used to evaluate changes in sleep and emotions of rats. The metabolic-related indicators such as blood pressure, blood viscosity, blood glucose, and uric acid in rats were measured. The pathological changes in the cornu ammonis 1 (CA1) region of rat brain were evaluated using hematoxylin and eosin staining and Nissl staining. Additionally, the sleep-related factors gamma-aminobutyric acid (GABA), glutamate (GA), 5-hydroxytryptamine (5-HT), and interleukin-6 (IL-6) were measured using enzyme linked immunosorbent assay. Finally, we screened potential sleep-improving receptors of DOE using polymerase chain reaction (PCR) array and validated the results with quantitative PCR and immunohistochemistry.
RESULTS:
DOE significantly improved rats' sleep and mood, increased the sodium pentobarbital-induced sleep time and sucrose preference index, and reduced autonomic activity times (P<0.05 or P<0.01). DOE also had a good effect on metabolic abnormalities, significantly reducing triglyceride, blood glucose, blood pressure, and blood viscosity indicators (P<0.05 or P<0.01). DOE significantly increased the GABA content in hippocampus and reduced the GA/GABA ratio and IL-6 level (P<0.05 or P<0.01). In addition, DOE improved the pathological changes such as the disorder of cell arrangement in the hippocampus and the decrease of Nissel bodies. Seven differential genes were screened by PCR array, and the GABAA receptors (Gabra5, Gabra6, Gabrq) were selected for verification. The results showed that DOE could up-regulate their expressions (P<0.05 or P<0.01).
CONCLUSION
DOE demonstrated remarkable potential for improving insomnia, which may be through regulating GABAA receptors expressions and GA/GABA ratio.
Animals
;
Dendrobium/chemistry*
;
Rats, Sprague-Dawley
;
Male
;
Sleep Initiation and Maintenance Disorders/blood*
;
Plant Extracts/therapeutic use*
;
Receptors, GABA-A/metabolism*
;
Noise/adverse effects*
;
Light/adverse effects*
;
gamma-Aminobutyric Acid/metabolism*
;
Sleep/drug effects*
;
Rats
;
Receptors, GABA/metabolism*
6.Clinical value of quality control circle activities in preventing deep vein thrombosis in military hospitals
Xile WEI ; Xuehui HU ; Hange SONG ; Meixia ZHANG ; Guang CHENG ; Heng LUO ; Desheng WANG
Journal of Navy Medicine 2025;46(2):150-155
Objective To explore the clinical effect of quality control circle(QCC)activities on the prevention of postoperative deep vein thrombosis(DVT)in inpatients in military hospitals.Methods A total of 318 patients who were diagnosed and treated in The First Affiliated Hospital of Air Force Medical University from January to December 2021 and 40 medical staff were enrolled in this study.Routine care was performed in 158 patients from January to June 2021,and QCC care was implemented in 160 patients from July to December 2021.The awareness of DVT prevention in medical staff and patients(or their famiy members)before and after QCC activities,lower limb DVT preventive measures taken by medical staff,and the occurrence of DVT were compared.Results The scores of the cause,clinical manifestations,nursing measures and preventive measures of DVT after QCC activities were significantly higher than those before QCC activities in both medical staff and patients(or their families)(P<0.05).The overall implementation rate of preventing lower limb DVT after QCC activities was significantly higher than that before QCC activities(94.14%vs.46.20%,P<0.05).The incidence of DVT after QCC activities was significantly lower than that before QCC activities(0.62%vs.5.06%,P<0.05).Conclusion Implementing QCC activities can improve the cognitive ability of military medical staff and patients(or family members)in DVT prevention,increase the implementation rate of DVT prevention measures,and reduce the incidence of DVT.
7.Production of GTKO pigs and kidney xenotransplantation from pigs to rhesus macaques
Yan WANG ; Yue CHANG ; Chang YANG ; Taiyun WEI ; Xiaoying HUO ; Bowei CHEN ; Jiaoxiang WANG ; Heng ZHAO ; Jianxiong GUO ; Hongfang ZHAO ; Xiong ZHANG ; Feiyan ZHU ; Wenmin CHENG ; Hongye ZHAO ; Kaixiang XU ; Ameen Jamal MUHAMMAD ; Zhendi WANG ; Hongjiang WEI
Organ Transplantation 2025;16(4):526-537
Objective To explore the construction of α-1,3-galactosyltransferase (GGTA1) gene-knockout (GTKO) Diannan miniature pigs and the kidney xenotransplantation from pigs to rhesus macaques, and to assess the effectiveness of GTKO pigs. Methods The GTKO Diannan miniature pigs were constructed using the CRISPR/Cas9 gene-editing system and somatic cell cloning technology. The phenotype of GTKO pigs was verified through polymerase chain reaction, Sanger sequencing and immunofluorescence staining. Flow cytometry was used to detect antigen-antibody (IgM) binding and complement-dependent cytotoxicity. Kidney xenotransplantation was performed from GTKO pigs to rhesus macaques. The humoral immunity, cellular immunity, coagulation and physiological indicators of the recipient monkeys were monitored. The function and pathological changes of the transplanted kidneys were analyzed using ultrasonography, hematoxylin-eosin staining, immunohistochemical staining and immunofluorescence staining. Results Single-guide RNA (sgRNA) targeting exon 4 of the GGTA1 gene in Diannan miniature pigs was designed. The pGL3-GGTA1-sgRNA1-GFP vector was transfected into fetal fibroblasts of Diannan miniature pigs. After puromycin selection, two cell clones, C59# and C89#, were identified as GGTA1 gene-knockout clones. These clones were expanded to form cell lines, which were used as donor cells for somatic cell nuclear transfer. The reconstructed embryos were transferred into the oviducts of trihybrid surrogate sows, resulting in 13 fetal pigs. Among them, fetuses F04 and F11 exhibited biallelic mutations in the GGTA1 gene, and F04 had a normal karyotype. Using this GTKO fetal pig for recloning and transferring the reconstructed embryos into the oviducts of trihybrid surrogate sows, seven surviving piglets were obtained, all of which did not express α-Gal epitope. The binding of IgM from the serum of rhesus monkey 20# to GTKO pig PBMC was reduced, and the survival rate of GTKO pig PBMC in the complement-dependent cytotoxicity assay was higher than that of wild-type pig. GTKO pig kidneys were harvested and perfused until completely white. After the left kidney of the recipient monkey was removed, the pig kidney was heterotopically transplanted. Following vascular anastomosis and blood flow restoration, the pig kidney rapidly turned pink without hyperacute rejection (HAR). Urine appeared in the ureter 6 minutes later, indicating successful kidney transplantation. The right kidney of the recipient was then removed. Seven days after transplantation, the transplanted kidney had good blood flow, the recipient monkey's serum creatinine level was stable, and serum potassium and cystatin C levels were effectively controlled, although they increased 10 days after transplantation. Seven days after transplantation, the levels of white blood cells, lymphocytes, monocytes and eosinophils in the recipient monkey increased, while platelet count and fibrinogen levels decreased. The activated partial thromboplastin time, thrombin time and prothrombin time remained relatively stable but later showed an upward trend. The recipient monkey survived for 10 days. At autopsy, the transplanted kidney was found to be congested, swollen and necrotic, with a small amount of IgG deposition in the renal tissue, and a large amount of IgM, complement C3c and C4d deposition, as well as CD68+ macrophage infiltration. Conclusions The kidneys of GTKO Diannan miniature pigs may maintain normal renal function for a certain period in rhesus macaques and effectively overcome HAR, confirming the effectiveness of GTKO pigs for xenotransplantation.
8.Ectopic thyroid gland in parotid region: a case report and literature review
ZHANG Lei ; CHEN Xibo ; HENG Wei ; WANG Xuefeng ; WANG Yangyang ; LIU Rui
Journal of Prevention and Treatment for Stomatological Diseases 2025;33(8):659-665
Objective:
To explore the clinical characteristics, diagnosis, and treatment of ectopic thyroid gland in the parotid gland area, and to provide clinical ideas for the diagnosis and treatment of ectopic thyroid gland.
Methods:
A case of a normal thyroid gland with ectopic thyroid gland tissue in the parotid gland area in the neck was reported. The male patient was 20 years old. The chief complaint was the discovery of a painless mass gradually increasing under the left earlobe for one month. Clinical examination showed obvious bulging of the tissue under the left earlobe. A strip-shaped mass approximately 3.0 cm long could be palpated. It was soft in texture, with a clear boundary, and located under the skin. The skin was pale red and of normal temperature. The body position movement test was negative. Color Doppler ultrasound of the thyroid gland in the neck showed that the shape and size of the thyroid gland were normal. CT images of the head and neck showed a band-like soft tissue density shadow at the area of the parotid gland behind and below the left earlobe, with a clear boundary. The CT value was approximately 30 HU, and further enhancement yielded no additional findings. The admitting diagnosis was a mass in the left parotid gland area. The tumor was incised using a conventional surgical method for the parotid gland area. During the operation, it was found that the tumor was located under the skin, and the contents were bright-red granulomatous tissue without a capsule and adhesive to the skin tissue. The parotid gland capsule was not involved. After the tumor was completely scraped off, intermittent suturing was performed. The resected tumor was sent for pathological examination. A retrospective analysis of the diagnosis and treatment of this type of case was conducted in combination with a literature review.
Results:
The wound of the patient failed to heal in the first stage after the operation. By applying iodoform gauze for pressurized dressing changed weekly, the wound gradually healed about 2 months later. The postoperative pathological report showed an ectopic thyroid gland in the left parotid gland area. The results of the literature review indicate that ectopic thyroid glands can be partial or complete. In the former, normal thyroid gland tissue exists in the neck, and some thyroid gland tissue appears in other locations, mostly at the base of the tongue and mediastinum. In the latter, the thyroid gland in the neck is absent. Both can present with abnormal thyroid gland function and local compression symptoms, and the symptoms are more obvious in patients with a complete ectopic thyroid gland. Ectopic thyroid glands are mainly diagnosed and differentiated through physical examination and imaging examination. Ectopic thyroid glands occurring subcutaneously in the parotid gland area are extremely rare. Physicians should design personalized treatment plans based on clinical examinations and surgical indications.
Conclusion
A subcutaneous ectopic thyroid gland in the parotid gland area is rare. For ectopic thyroid gland surgery, a reasonable surgical plan should be designed considering the patient's aesthetic needs and prognosis. Puncture biopsy should be performed when necessary to formulate the surgical plan.
9.Construction and Functional Validation of GTKO/hCD55 Gene-Edited Xenotransplant Donor Pigs
Jiaoxiang WANG ; Lu ZHANG ; Shuhan CHEN ; Deling JIAO ; Heng ZHAO ; Taiyun WEI ; Jianxiong GUO ; Kaixiang XU ; Hongjiang WEI
Laboratory Animal and Comparative Medicine 2025;45(4):379-392
Objective To develop GTKO (α-1,3-galactosyltransferase gene-knockout, GTKO)/hCD55 (human CD55) gene-edited xenotransplant donor pigs and verify their function. Methods In this study, CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated nuclease 9), PiggyBac transposon technology and somatic cell nuclear transfer technology were used to construct GTKO/hCD55 gene-edited Diannan miniature pigs. The phenotype and function of GTKO/hCD55 pigs were analyzed by Sanger sequencing, real-time fluorescence quantitative PCR, flow cytometry, immunofluorescence, bisulfite sequencing, antigen-antibody binding assays, and complement-dependent cytotoxicity assays. Results After transfection of PX458 and PiggyBac gene editing vectors into wild-type fetal pig fibroblasts, 48 single-cell colonies were obtained through puromycin drug screening. Two single-cell colonies were selected for somatic cell nuclear transfer, resulting in two fetal pigs at 33 days of gestation. The GGTA1(α-1,3-galactosyltransferase) genotypes of fetal pig F01 were -17 bp and wild type (WT), while the GGTA1 genotypes of fetal pig F02 were -26 bp/+2 bp and -3 bp. The hCD55 mRNA expression levels of both fetal pigs were significantly higher than those of WT pigs (P<0.01). The fetal pig F02 was selected as the donor cell source for recloning, 11 surviving piglets were obtained, all identified as GTKO/hCD55 gene-edited pigs. These pigs showed absence of α-Gal antigen expression, but weak or no expression of hCD55 was observed. Methylation analysis of the hCD55 gene's CpG island showed hypermethylation in kidney tissue lacking hCD55 expression, whereas it was not methylated or partially methylated in kidney tissue expressing hCD55. Moreover, codon optimization of the CpG island of the hCD55 gene to reduce CG content could achieve stable expression of the hCD55 gene. In addition, antigen-antibody binding experiment showed that the amount of human IgM binding to GTKO/hCD55 gene-edited pig fibroblasts was significantly lower than that of WT pigs (P<0.01). Complement-dependent cytotoxicity experiment showed that the survival rate of fibroblasts in GTKO/hCD55 pigs was significantly higher than that in WT pigs (P<0.01). Conclusion This study demonstrates the successful generation of GTKO/hCD55 gene-edited xenotransplant donor pigs. Methylation-induced gene silencing of the hCD55 gene can be effectively avoided by reducing the CG content of the CpG island through codon optimization. This study provides a reference for the development of xenotransplant donor pigs and guides subsequent research on xenotransplantation.
10.Differences of perineuronal nets and parvalbumin-positive neuron distribution across specific brain regions in young and aged mice
Mei FU ; Wei-Yi YE ; Heng-Hao HOU ; Qian CHEN
Acta Anatomica Sinica 2025;56(5):515-523
Objective To investigate the differences of perineuronal net(PNN)and parvalbumin(PV)-positive neuron distribution across specific brain regions between young and aged mice.Methods Brains from young(45 days)and aged(350 days)mice(n=4 per group)were fixed with 4%paraformaldehyde,sectioned(50 μm)using a vibratome,and stained with Wisteria floribunda agglutinin(WFA)and PV immunofluorescence.Quantitative analyses of PNN-positive and PV-positive neurons,along with PNN encapsulation of PV-positive neurons,were performed in the anterior cingulate cortex(ACC),somatosensory cortex barrel field layer 4(S1BF L4),striatum(STR),and hippocampal CA2 region.Results Aged mice exhibited no significant changes in PNN-positive or PV-positive neuron counts in ACC,S1BF L4,or STR compared to young mice,but showed significantly increased PNN encapsulation of PV-positive neurons.In hippocampal CA2,PNN-positive neurons increased significantly without PV-positive neuron alterations.Conclusion The differences in PNN-PV neuron interactions and PNN density exist in specific brain regions of young and aged mice.


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