Human naïve pluripotent stem cells (PSCs) hold great promise for embryonic development studies. Existing induction and culture strategies for these cells, heavily dependent on MEK inhibitors, lead to widespread DNA hypomethylation, aberrant imprinting loss, and genomic instability during extended culture. Here, employing high-content analysis alongside a bifluorescence reporter system indicative of human naïve pluripotency, we screened over 1,600 chemicals and identified seven promising candidates. From these, we developed four optimized media-LAY, LADY, LUDY, and LKPY-that effectively induce and sustain PSCs in the naïve state. Notably, cells reset or cultured in these media, especially in the LAY system, demonstrate improved genome-wide DNA methylation status closely resembling that of pre-implantation counterparts, with partially restored imprinting and significantly enhanced genomic stability. Overall, our study contributes advancements to naïve pluripotency induction and long-term maintenance, providing insights for further applications of naïve PSCs.
Humans ; DNA Methylation/drug effects* ; Genomic Instability ; Pluripotent Stem Cells/metabolism* ; Cell Culture Techniques/methods* ; Cells, Cultured

OBJECTIVE: To assess the practical and health economical values of non-invasive prenatal test (NIPT) in Changsha Municipal Public Welfare Program. METHODS: A retrospective analysis was carried out on 149 165 women undergoing NIPT test from April 9, 2018 to December 31, 2019. For pregnant women with high risks, invasive prenatal diagnosis and follow-up of pregnancy outcome were conducted. The cost-benefit of NIPT for Down syndrome was analyzed. RESULTS: NIPT was carried out for 149 165 pregnant women and succeeded in 148 749 cases (99.72%), for which outcome were available in 148 538 (99.86%). 90% of pregnant women from the region accepted the screening with NIPT. 415 (0.27%) were diagnosed as high risk. Among these, 381 (91.81%) accepted amniocentesis, which led to the diagnosis of 212 cases of trisomy 21 (PPV=85.14%), 41 cases with trisomy 18 (PPV=48.81%) and 10 cases with trisomy 13 (PPV=20.83%). The sensitivity and specificity of NIPT for trisomy 21, trisomy 18 and trisomy 13 were (97.70%, 99.98%), (97.62%, 9.97%) and (100%, 99.97%), respectively. In addition, 213 and 30 cases were diagnosed with sex chromosomal aneuploidies (PPV=46.2%) and other autosomal anomalies (PPV=16.57%), respectively. For Down syndrome screening, the cost and benefit of the project was 120.79 million yuan and 1,056.95 million yuan, respectively. The cost-benefit ratio was 1: 8.75, and safety index was 0.0035. CONCLUSION NIPT is a highly accurate screening test for trisomy 21, which was followed by trisomy 18 and sex chromosomal aneuploidies, while it was less accurate for other autosomal aneuploidies. The application of NIPT screening has a high health economical value.
Aneuploidy ; Cost-Benefit Analysis ; Female ; Humans ; Noninvasive Prenatal Testing ; Pregnancy ; Retrospective Studies ; Trisomy 18 Syndrome/genetics*

OBJECTIVE: To explore the analgesic mechanism of intrathecal trichostatin A (TSA) injection in a rat model of neuropathic pain induced by chronic constrictive injury (CCI). METHODS: Male SD rats were randomized into sham operation+ DMSO group (group S), CCI +DMSO group (group C), CCI +10 μg TSA group (group T), and in the latter two groups, rat models of neuropathic pain were established induced by CCI. The rats were given intrathecal injections of 10 μL 5% DMSO or 10 μg TSA (in 5% DMSO) once a day on days 7 to 9 after CCI or sham operation. The rats were euthanized after behavioral tests on day 10, and the lumbar segment of the spinal cord was sampled to determine the expression of histone deacetylase 4 (HDAC4) protein and mRNA and detect the differentially expressed miRNAs using a miRNA chip. MiR-190b-5p and miR-142-3p were selected for validation of the results using RT-qPCR. RESULTS: Compared with those in group S, the rats in group C showed significantly decreased paw withdrawal mechanical threshold (PWMT) from day 3 to day 10 after CCI ( < 0.05); intrathecal injection of TSA significantly reversed the reduction of PWMT following CCI ( < 0.05). Positive HDAC4 expression was detected mainly in the cytoplasm of the neurons in the gray matter of the spinal cord, and was obviously up-regulated after CCI ( < 0.05). Intrathecal injection of TSA significantly suppressed CCI-induced up-regulation of HDAC4 at 10 days after the operation ( < 0.05). Compared with the miRNA profile in group S, miRNA profiling identified 83 differentially expressed miRNAs in group C (fold change ≥2 or ≤0.5, < 0.05); TSA treatment reversed the expressions of 58 of the differentially expressed miRNAs following CCI, including 41 miRNAs that were decreased after CCI but up-regulated following TSA treatment. The results of real-time PCR validated the changes in the expressions of miR-190b-5p and miR-142-3p. CONCLUSIONS TSA suppresses CCI-induced up-regulation of HDAC4 and reverses differential expressions of miRNAs in the spinal cord of rats, which may contribute to the analgesic effect of TSA on neuropathic pain.
Animals ; Histone Deacetylases ; Hydroxamic Acids ; Male ; MicroRNAs ; Rats ; Rats, Sprague-Dawley ; Spinal Cord ; Up-Regulation

Objective To study whether preoperative serum thyrotropin ( TSH) concentration can be used for risk prediction of papillary thyroid microcarcinoma ( PTMC ) . Methods The cohort of this retrospective study consisted of 1 707 patients who underwent surgery on thyroid nodules at Chinese PLA General Hospital from October 1999toFebruary2011. 37.32%(n=637)ofthesepatientssufferedfromdifferentiatedthyroidcancer(DTC),and 14. 18%(n=242) of patients with DTC suffered from PTMC. Results (1) The mean TSH level in patients with DTC was significantly higher than that in patients with benign thyroid nodules [(1. 99(1. 25-3. 19) vs 1. 48 (0. 85-2. 32) mU/L, P<0. 01]. DTC with diameter greater than 10 mm had higher serum TSH level compared with that in benign thyroid nodules[2. 04(1. 26-3. 36) vs 1. 45(0. 83-2. 30), P<0. 01]. Serum TSH level was not significantly raised in cases where-as the diameter of tumor was 10 mm or less. (2) With the increasing level of TSH, the prevalence of DTC and tumours with diameter greater than 10 mm rose significantly, but the increasing trend was not significant in PTMC. (3) Raised TSH level was an independent risk factor of DTC based on Binary logistic regression. Conclusions Serum TSH is an independent risk predictor of DTC, it is an independent risk predictor of the diameter of DTC greater than 10 mm, but it is not a good risk predictor in PTMC.

Objective To study gender-related differences of clinical characteristics and vascular complications in patients with aldosterone-producing adenoma(APA).Method Consecutive 315 patients with APA confirmed by pathological diagnosis were included and the clinical features and vascular complications were compared based on gender.Results (1) Of the 315 patients with APA,female accounted for 52.7％ (166/315).Male patients with APA presented a higher BMI,compared with females [(25.4± 2.9) vs (24.1 ± 3.2) kg/m2,P＜0.01],and the history of smoking and drinking was more common in male patients.No significant difference was found in regard to the age,duration,hypokalemia,tumor size,and family history of hypertension between two groups (P＞0.05).(2) No significant difference was found in the prevalence of hypertension,duration of hypertension,blood pressure,and antihypertensive medieation between two groups.However,the patients with grade 3 hypertension tended to be more prevalent in males(P =0.08).(3) Serum aldosterone concentrations were similar in two groups,but 24 h urinary aldosterone showed an increased trend in male group (P =0.07).(4) The overall prevalences of cardiovascular (51.0％ vs 36.1％) and cerebrovascular (9.4％ vs 3.0％) complications were significantly higher in male group (P ＜ 0.05).Further analysis of cardiac events revealed significantly higher rates of left ventricular hypertrophy (23.5％ vs 13.9％) and arrhythmia (21.5％ vs 10.8％) in males (P＜0.05).However,no significant difference in the prevalence between two groups was found in regard to cerebral hemorrhage,infarction,and chronic renal insufficiency.(5) The fall of blood pressure and recovery from hypokalemia were comparable between males and females in two weeks after sugery.However,in the patients with persistent hypertension,the number of antihypertensive drugs used in males was greater than that in females and more male patients needed ≥ 2 types of antihypertensive drugs.Conclusion There were significant gender-related differences with regard to clinical features and vascular complications in patients with APA.

A 31-year-old male normotensive patient with aldosterone-producmg adenoma complained of thirst,polydipsia,polyuria,and periodical paraplegia.The diagnosis is raised by signs of hypokalemia.Despite the lack of hypertension,primary aldosteronism was confirmed by persistent hypokalemia,increased urinary potassium,increased urinary and plasma aldosterone levels and suppressed plasma rennin activity (PRA).The blood pressure profile was studied by ambulatory monitoring,and the mean blood pressure of 24h was normal and the circadian rhythm remained normal. Surgical removal of the histologically typical aldosterone-producing adenomas normalized the kalemia.The patient had a marked fall in blood pressure with mean values of 21/17 mm Hg ( diurnal and nocturnal blood pressure were 19/17 and 22/17 mm Hg respectively)and recovery of normal urinary and plasma aldosterone levels and PRA 6 weeks after surgery.This suggests that excess serum aldosterone induced relative hypertension in those patients whose blood pressure was spontaneously very low.Our observations call for primary hyperaldosteronism assay in patients with hypokalemia and renal potassium leakage.

Objective To better understand the clinical management of tumor-induced osteomalacia (TIO) by analyzing the clinical features, diagnosis, treatment, postoperative biochemical changes, and clinical status in 12 cases of TIO. Methods Twelve cases of TIO hospitalized from 2004 to April 2010 were reviewed retrospectively. All cases were diagnosed based on their clinical manifestation, hypophosphatemia, and image study including technetium-99m octreotide scintigraphy (99mTc-Oct). Resuits There were 7 males and 5 females with mean age of (41.8±9.6) years (20 to 56 years). The course of disease was from 2 to 14 years ( median course 4.0 years). They all presented with bone pain, gait disturbance, muscle pain, and muscle weakness. Serum phosphate( Pi)levels were low in 12 cases with a range from 0.30 to 0.56 mmol/L. 99mTc-Oct was performed in 9 cases and it showed that the lesions were located in head of femur, fibula, retrocalcaneal area, foot, humerus,metacarpal, posterior chest wall or near nasal bone (apex partis petrosae ossis temporalis). Subcutaneous soft tissue mass was found in another 3 cases at loin, thigh, and foot by physical examination. The tumors were confirmed by CT, MRI or ultrasonography. Twelve patients underwent operation to remove the tumors and histopathology showed hemangioendothelioma or fibrous angioma (6 cases), giant cell tumor or fibroma of tendon sheath(4 cases), liposarcoma(1case), and phosphaturic mesenchymal tumor(1case). Serum Pi levels returned to normal in 10 patients after resection of tumor. During 2 to 64 months follow up, symptoms of bone pain and muscle weakness were improved obviously. Conclusions Patients with hypophosphatemic osteomalacia should be thoroughly investigated for TIO. 99mTc-Oct and other imaging examinations can effectively locate the tumors. Once the hidden tumor is found and excised, the patient will recover and enjoy normal life with normalized Pi concentrations and marked improvement of symptoms.

Objective To analyze the changes of heart rate turbulence (HRT) and heart rate variability (HRV) in coronary heart disease (CHD), and evaluate the relationship between CHD and coronary artery disease (CAD) and two parameters of HRT and HRV, and investigate the correlation between the 2 parameters.Methods We performed coronary angiography in all 103 patients with CHD (group A), who were classified into 3 subgroups: 35 patients of stable angina pectoris ( group A1 ), 28 patients of unstable angina pectoris ( group A2 ) and 40 patients of AMI ( group A3 ), and 30 structurally normal patients with ventricular premature contraction who were included in healthy control group ( group B).The 24-hour ambulatory electrocardiogram was performed in all the patients, and the parameters of HRT and HRV were obtained.Results The value of the TO onset of group A was significantly higher than that of group B( -4.17± 2.75, - 3.16 ± 3.18, - 0.96 ± 2.92; - 6.30 ± 3.47 ), the value of TS of group A was lower than that of groupB(6.81±3.18,5.12±3.31,3.20±1.71;9.52±3.85) (P ＜0.05 orP ＜0.01).The values of SDNN, PNN50, and HF of group A were significantly lower than those of group B (P＜ 0.05).TO was positively correlated with Gensinies score, while TS, SDNN, PNN50 and HF were negatively correlated with Gensinies score, and TS showed the strongest negative correlation with Gensini score (r=-0.45).TO was negatively correlated with SDNN, PNN50, and HF, while TS was positively correlated with SDNN, PNN50 and HF, and TS showed much stronger correlation with HF (r=0.47).Conclusions HRT was dramatically blunted and HRV in patients with CHD was significantly lower;.HRT and HRV of group A1 were significantly correlated with Gensini score, and TS had the strongest correlation with Gensini score.TO and TS were correlated with SDNN, PNN50 and HF, and TS had much stronger relationship with HF.

Objective To establish a method for determining the content of Naringin and Hesperidin in Zengshi Keli by HPLC. Methods The Kromasil C18 column with acetonitrile-water-phosphoric acid (20∶80∶0.02) as the mobile phase was used. The flow rate was 1.0 mL/min, the detective wavelength was 283 nm, the temperature of column is 35 ℃. Results The calibration curve were linear in the range of 0.178 5～0.892 5 ?g for Naringin and 0.073 68～0.368 4 ?g for Hesperidin (r=0.999 9) respectively. The average recovery was 97.24% (RSD=1.21%) and 96.95% (RSD=1.49%) respectively. Conclusion The method was simple, accurate, reproducible and can be used for quality control of Zengshi Keli.

Objective To study the relationship between HCMV infection and endothelial function disorder and its possible role in CHD and acute coronary syndrome (ACS).Method HCMV-DNA, IgM, IgG, ET, sICAM were detected in 59 patients with CHD(CHD group) and 27 patients without CHD(non-CHD group ). The CHD group was divided into acute coronary syndrome group (group A,n=35) and stable symptom group (group B,n=24).Results The positive rate of HCMV-DNA, IgM, IgG was higher in CHD group, especially in group A than that in non-CHD group (P