OBJECTIVES: The core pathology of osteoporosis lies in bone resorption exceeding bone formation; thus, promoting osteogenesis is a key therapeutic strategy. The osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) forms the biological basis of bone formation. Astragaloside IV (A-IV), a major active component of Astragalus membranaceus, is known to enhance osteogenesis, but its precise molecular mechanisms remain unclear. This study aims to investigate the effects of A-IV on the proliferation and osteogenic differentiation of BMSCs from osteoporotic rats and to elucidate its molecular mechanism through the regulation of microRNA-21 (miR-21) and Notch2 expression. METHODS: After 1 week of adaptive feeding, mature female SD rats were randomly divided into a sham-operated (Sham) group (n=4) and an ovariectomized (OVX) group (n=8) to establish an osteoporosis model. Twelve weeks after surgery, BMSCs were isolated from femoral bone marrow and cultured. Cells were divided into a S-BMSCs group (from Sham), an O-BMSCs group (from OVX), and an A-BMSCs group (from OVX-derived BMSCs treated with A-IV). S-BMSCs and O-BMSCs were induced for osteogenic differentiation using osteogenic induction medium, whereas A-BMSCs were treated with A-IV before induction. Flow cytometry was used to identify mesenchymal stem cell surface markers (CD29) and hematopoietic stem cell marker (CD34) to confirm BMSC characteristics. Cell proliferation was assessed using the methyl thiazolyl tetrazolium (MTT) assay. Alizarin red staining was performed to quantify calcium nodule formation, and alkaline phosphatase (ALP) activity assays were used to evaluate osteogenic differentiation. Real-time reverse transcription PCR (real-time RT-PCR) was used to detect changes in osteogenic-related genes, runt-related transcription factor 2 (Runx2) and osteopontin (OPN), as well as miR-21 expression. Western blotting was performed to assess Runx2, OPN, and Notch2 protein expression. RESULTS: Flow cytometry confirmed that O-BMSCs retained the phenotypic characteristics of mesenchymal stem cells. A-IV significantly enhanced the proliferation of BMSCs from osteoporotic rats (P<0.05), increased ALP activity, and upregulated the mRNA and protein expression of Runx2 and OPN (P<0.05). Bioinformatic and experimental analyses demonstrated that miR-21 directly targeted Notch2. A-IV treatment increased miR-21 expression while suppressing Notch2 protein expression and inhibiting activation of the Notch signaling pathway (P<0.05). CONCLUSIONS Astragaloside IV promotes the osteogenic differentiation of BMSCs derived from osteoporotic rats by upregulating miR-21 expression and inhibiting the key Notch signaling protein Notch2, thereby relieving the Notch2-mediated suppression of osteogenesis.
Animals ; Triterpenes/pharmacology* ; Saponins/pharmacology* ; Osteogenesis/drug effects* ; MicroRNAs/metabolism* ; Rats, Sprague-Dawley ; Female ; Cell Differentiation/drug effects* ; Mesenchymal Stem Cells/drug effects* ; Signal Transduction/drug effects* ; Osteoporosis/pathology* ; Rats ; Cells, Cultured ; Receptor, Notch2/metabolism* ; Receptors, Notch/metabolism* ; Ovariectomy ; Cell Proliferation/drug effects*

OBJECTIVES: To explore the efficacy of DSA-guided intrathecal drug delivery system combined with Zi Wu Liu Zhu Acupoint Therapy for management of cancer pain and provide reference for its standardized clinical application. Methods and. RESULTS: Recommendations were formulated based on literature review and expert group discussion, and consensus was reached following expert consultation. The consensus recommendations are comprehensive, covering the entire treatment procedures from preoperative assessment and preparation, surgical operation process, postoperative management and traditional Chinese medicine treatment to individualized treatment planning. The study results showed that the treatment plans combining traditional Chinese with Western medicine effectively alleviated cancer pain, reduced the use of opioid drugs, and significantly improved the quality of life and enhanced immune function of the patients. Postoperative follow-up suggested good treatment tolerance among the patients without serious complications. CONCLUSIONS The formulated consensus is comprehensive and can provide reference for clinicians to use DSA-guided intrathecal drug delivery system combined with Zi Wu Liu Zhu Acupoint Therapy. The combined treatment has a high clinical value with a good safety profile for management of cancer pain.
Humans ; Medicine, Chinese Traditional ; Cancer Pain/therapy* ; Drugs, Chinese Herbal/therapeutic use* ; Drug Delivery Systems ; Pain Management/methods* ; China

Objective To analyze the detection efficiency of p16INK4a protein combined with human papillomavirus and liquid-based cytology(LCT)in the screening of cervical precancerous lesions,and to provide a basis for cervical cancer preven-tion and treatment.Methods The results of p16INK4a staining of cervical epithelial cells,human papillomavirus testing and cer-vical cytology were analyzed in 139 inpatients at Guangzhou Women's and Children's Medical Center between January 2019 and December 2020.Of them,there were 111 patients with cervical intraepithelial neoplasia(CIN)and 28 cases of cervical inflam-matory disease.The efficacy of the three methods alone and in combination to screen for CIN lesions was compared.Results In the detection of CIN patients,the sensitivity of p16INK4a,microfluidic microarray and cervical cytology for detecting CIN and a-bove lesions was 91.89% ,94.59% and82.88% ,with specificity of 57.14% ,17.86% and46.43% ,and AUC of 0.75,0.56 and 0.65,respectively;while the sensitivity of"p16INK4a+LCT","p16INK4a+hrHPV","LCT+hrHPV"and their sen-sitivity were 96.40% ,97.30% ,94.59% and 99.10% ,their specificity was 85.71% ,92.86% ,89.29% and 92.86% ,and the AUC was 0.91,0.95,0.92 and 0.96,respectively.Conclusion The combined p16INK4a and hrHPV test helps to improve diagnostic accuracy and early detection,thus allowing for earlier intervention or treatment.This combined application allows for more accurate identification of low-grade and high-grade cervical intraepithelial neoplasia,providing more information for indi-vidualized patient management.

ObjectiveThe human angiotensin converting enzyme2 （hACE2） transgenic mouse model was used to clarify the antiviral efficacy of BD-77 against a novel coronavirus SARS-CoV-2 and explore the action mechanism of BD-77 against SARS-CoV-2. MethodSARS-CoV-2 Omicron and Delta variant strains-infected VeroE6 cell models were established and administered with BD-77 to observe the antiviral effect of BD-77 in vitro. A kit was used to detect the effect of BD-77 in vitro on the binding of spike S protein of SARS-CoV-2 virus （Delta/Omicron） to angiotensin converting enzyme2 （ACE2）. Chromatography was adopted to detect the binding of BD-77 to the S protein and N protein of the novel coronavirus. hACE2 transgenic C57BL/6 mice were divided into a blank control group， SARS-CoV-2 infection group， BD-77 administration groups of 37.5 mg·kg^-1 and 75 mg·kg^-1， with eight mice in each group. The pneumonia model of SARS-CoV-2-infected hACE2 transgenic mice was built to observe the survival of the mice， detect the virus titer of the lung tissue of the mice， and observe the lesions in the lung tissue. ResultBD-77 had a certain inhibitory effect on Omicron and Delta variant strains in vitro， with median inhibitory concentration （IC₅₀） of 526.3 mg·L^-1 and 653.0 mg·L^-1， respectively. BD-77 had no significant inhibitory effect on the binding of the S protein of WT， Omicron， and Delta variant strains of SARS-CoV-2 to ACE2 and had no binding effect with the S protein and N protein of the novel coronavirus. No mice in the blank group died， while the mortality rate of SARS-CoV-2-infected mice was 75%. There was a large amount of virus replication in the lung tissue of the mice and large areas of inflammatory infiltration in the lung tissue and interstitium. Compared with the model group， BD-77 administration groups of 37.5 mg·kg^-1 and 75 mg·kg^-1 could reduce the mortality of mice， significantly lower the virus titer in the lung tissue of mice （P<0.05）， and improve lung lesions. ConclusionBD-77 demonstrated significant inhibitory effects against SARS-CoV-2 virus in vitro and in vivo. However， its mechanism of action did not involve direct inhibition of the virus itself or intervention in the virus-host binding process. This finding suggests that the mechanism of action of BD-77 needs to be thoroughly investigated and elucidated by further experiments.

Objective To investigate the efficacy and safety of a single induction dose of cyclopofol in painless four-dimensional hysterosalpingography,this study aims to assess its academic significance and adherence to Nature journal's publication standards.Methods A total of 120 patients admitted to Guangdong Reproductive Hospital between February 2022 and December 2022,requiring painless vaginal four-dimensional ultrasound hysterosalpin-gography,were selected as the study population.Random selection was performed with 40 patients in each group.The dosage of cyclopofol administered was 0.3 mg/kg(C1 group),0.5 mg/kg(C2 group),and 0.7 mg/kg(C3 group).Heart rate(HR),mean arterial pressure measured non-invasively using a cuff,time to wake after surgery,and incidence of adverse reactions were observed at three time points:before examination(T1),during contrast agent injection(T2),and after examination(T3).Results There were no significant differences in heart rate(HR)and mean arterial pressure(MAP)between T1 and T3 among the three groups(P>0.05).However,at T2,group C1 exhibited significantly higher HR and MAP compared to group C2(P<0.05),while there was no statistically significant difference in HR and MAP between group C2 and C3 at T2(P>0.05).The duration of wakefulness was shorter in group C2 than in group C3(P<0.05),but there was no statistical significance in wakefulness duration between group C1 and C2(P>0.05).The incidence of adverse reactions was lower in group C2(10.0%)compared to both group C1(32.5%)and group C3(37.5%)with statistical significance(P<0.05).Conclusion The admin-istration of cyclopofol at a dosage of 0.5 mg/kg exhibits notable advantages in painless four-dimensional hysterotubal ultrasonography,including effective sedation,prompt wakefulness,and a low occurrence rate of adverse reactions.

Objective:To compare the results of invasive prenatal diagnosis and preimplantation genetic testing (PGT) and explore the underlying mechanism.Methods:Clinical data of pregnant women undergoing PGT and invasive prenatal diagnosis at the Sixth Affiliated Hospital of Sun Yat-sen University from January 2019 to December 2022 were collected. The results of PGT and invasive prenatal diagnosis were compared, and the outcomes of pregnancies were followed up. This study has been approved by the Medical Ethics Committee of the the Sixth Affiliated Hospital of Sun Yat-sen University (No. 2022SLYEC-491).Results:A total of 172 couples were included in this study, and 26 non-targeted variants were discovered upon prenatal diagnosis, including 10 cases (38.5%) by chromosomal karyotyping, 15 (57.7%) by chromosomal microarray analysis (CMA), and 1 (3.8%) by whole exome sequencing. The 10 karyotypic anomalies had included 6 chromosomal polymorphisms, 2 chromosomal mosaicisms, 1 paternally derived translocation, and 1 missed maternal chromosomal inversion. CMA has identified 15 copy number variations (CNVs), which included 11 microdeletions and microduplications, 3 loss of heterozygosity, and 1 low-level mosaicism of paternal uniparental disomy. One CNV was classified as pathogenic, and another one was likely pathogenic, whilst the remaining 13 were classified as variants of uncertain significance. Therefore, 8.7% of CNVs was detected by invasive prenatal diagnosis after PGT. 92.3% (24/26) of the non-targeted variants have been due to technological limitations of next-generation sequencing (NGS).Conclusion:Invasive prenatal diagnosis after PGT can detect non-targeted variants, which may further reduce the incidence of birth defects.

OBJECTIVE The abnormal amyloid-β(Aβ)and oxidative stress assiociated with the progression of Alzheimer disease(AD).Quercetin has been reported to possess antioxidant and anti-inflammatory properties in neurodegenerative disorders.In this present study,we designed to characterize the mechanisms by which quer-cetin exerts neuroprotective effects in murine neuroblas-toma N2a cells stably expressing human Swedish mutant amyloid precursor protein(N2a/APP).METHODS N2a/APP cells were treated with quercetin at concentrations of 10,20 and 50 μ mol·L-1 for 24 h.Cell viability was examined with CCK-8 assays.The protein levels of ERK1/2 and Akt were detected by Western blotting.Intra-cellular reactive oxygen species(ROS)was detected by a fluorescent probe 2,7-dichlorofluorescein diacetate(DCFH-DA).The mitochondrial membrane potential(Δψ m)in N2a/APP cells was detected by using JC-1 staining method.Immunofluorescence was used to detect the generation of 8-hydroxy-2′-deoxyguanosine(8-OHdG)and 4-hydroxynonenal(4-HNE).RESULTS Quercetin attenuated the enhancement of p-ERK1/2,reductions of p-Akt,and decreased levels of APP expression.More-over,quercetin alleviated loss of mitochondria membrane potential(MMP)since it attenuates these oxidative stress,as reflected in the levels of ROS,4-HNE and 8-OHdG,was elevated in N2a/APP cells and these effects were again ameliorated by quercetin.CONCLUSION Neuroprotection by quercetin in N2a/APP cells involves normalizing the impaired mitochondrial function and reducing oxidative stress via inactivation of the ERK1/2 and activation of the Akt pathways.

Objective:To identify the effective concentrations of Ropivacaine in the modified fascial iliac compartment block(FICB)that would not affect the movement of the affected limb but would offer effective pain relief after total knee arthroplasty(TKA)in elderly patients.In addition, adverse reactions within 24 hours of FICB were examined.Methods:This study was a prospective, single-arm sequential trial.Forty-five elderly patients treated with TKA at the First Affiliated Hospital of Soochow University between September 2021 and March 2022 were selected, with an American Society of Anesthesiologists(ASA)score of Ⅰ or Ⅱ.All patients were given ultrasound-guided FICB on the surgical side under general anesthesia and 10 minutes before the operation, and the injection volume was 30 ml.According to preliminary experiments and relevant literature, the initial concentration of Ropivacaine was 0.1%, and the concentration for the next patient was determined using a modified Dixon sequential method.If the quadriceps femoris muscle strength score of the first patient was ≥4, there was no resting pain[visual analogue scale(VAS)score ≤1], and the VAS score during activity was ≤3, the concentration for the next patient would be reduced.Conversely, the concentration would be increased.The Ropivacaine concentration was increased or decreased by 0.01% each time and the trial would be stopped after 12 reentries.The median effective concentration(ED50), 95% effective concentration(ED95)and corresponding 95% confidence interval(CI)of ropivacaine were calculated using the probit model.Meanwhile, adverse reactions within 24 hours of FICB were monitored.Results:Of 43 elderly patients who completed the trial, the intervention was effective in 23 and ineffective in 20.While ensuring that postoperative limb movement in elderly patients was not affected, a single injection of 30 ml ropivacaine through FICB was able to effectively inhibit postoperative pain, and the ED50 and ED95 of ropivacaine were 0.072%(95% CI: 0.065%-0.078%)and 0.093%(95% CI: 0.084%-0.124%), respectively.Within 24 hours of FICB, 2 patients had lower limb weakness and could not get out of bed and walk, and 5 patients had severe pain and needed additional analgesics.No other adverse reactions were found. Conclusions:The effective ED50 and ED95 of Ropivacaine for postoperative pain relief in elderly patients after TKA are 0.072% and 0.093%, respectively.And the incidence of adverse reactions is low.

Objective:To investigate the value of short-time transcutaneous carbon dioxide pressure (TcPCO 2) and transcutaneous oxygen pressure (TcPO 2) monitoring in critically ill preterm infants. Methods:From January to December 2018, 62 critically ill neonates receiving respiratory support at Guangzhou Women and Children's Medical Center were retrospectively enrolled. A total of 348 sets of paired data including TcPCO 2/TcPO 2 and arterial carbon dioxide pressure (PaCO 2)/arterial oxygen partial pressure (PaO 2) were analyzed. The patients were divided into different groups based upon birth weight (23 cases>1 000 g-≤1 500 g, 129 sets of paired data; 18 cases≤1 000 g, 130 sets of paired data) and gestational age (16 cases born at ≤28 gestational weeks, 127 sets of paired data; 29 cases born at 28-34 gestational weeks, 159 sets of paired data) and the differences between groups were compared. The correlation and consistency of TcPCO 2/TcPO 2 and PaCO 2/PaO 2 were evaluated using Pearson's correlation and Blan-Altman scatter plots. Receiver operating characteristic (ROC) curve was drawn to analyze the diagnostic efficacy of TcPCO 2 in neonates with hypercapnia. Results:There was a positive correlation between TcPCO 2 and PaCO 2 in all patients ( r=0.913, 95% CI:0.894-0.929, P<0.05). In patients whose birth weight was>1 000 g-≤1 500 g or≤1 000 g, TcPCO 2 and PaCO 2 were positively correlated and the consistency were good ( r=0.909, 95% CI:0.874-0.935; r=0.934, 95% CI:0.908-0.953; both P<0.05), and the same finding was also observed in patients born at≤28 gestational weeks or 28-34 weeks of gestation ( r=0.938, 95% CI:0.913-0.956; r=0.871, 95% CI: 0.827-0.904; both P<0.05). The sensitivity, specificity and area under curve of TcPCO 2 in the diagnosis of hypercapnia were 90.91%, 85.85%, and 0.942, respectively. There was a poor correlation between TcPO 2 and PaO 2 in all patients and those with birth weight >1 000 g-≤1 500 g or gestational age 28-34 weeks (all r<0.75, all P<0.05). There was no correlation between TcPO 2 and PaO 2 in the birth weight ≤1 000 g and gestational age ≤28 weeks groups (both P>0.05). Conclusions:Short-time TcPCO 2 monitoring can accurately assess PaCO 2 in critically ill neonates requiring respiratory support and is of high diagnostic value for hypercapnia. However, TcPO 2 has limitation in evaluating PaO 2 and other indicators may need to be involved.

Objective:To evaluate the efficacy and safety of autologous hematopoietic stem cell transplantation(auto-HSCT)for the treatment of multiple myeloma(MM)in elderly patients aged over 65 years.Methods:In this retrospective analysis, the efficacy and safety of auto-HSCT for the treatment of MM was examined in 28 patients aged >65 years diagnosed and treated at Soochow Hopes Hematology Hospital between March 1, 2020 and October 31, 2022.The functions of the major organs of these patients were evaluated before transplantation.Results:The 28 patients had a median age of 67(66-72)at the time of transplantation, a median number of 2.985 × 10 6/kg(2.036-9.5 × 10 6/kg)of collected CD34+ hematopoietic stem cells, and a median number of 2(1-3)days of collection.The median time to neutrophil implantation after hematopoietic stem cell transfusion was 10(9-14)days, and the median platelet implantation time was 11(10-29)days.The median follow-up time was 25 months, but the median progression-free survival time was not reached.The 1-year PFS rate was 89.3% and 2-year PFS rate was 76.3%, with 2 patients' starting point of PFS set at the time of pre-transplantation re-induction therapy because needed salvage auto-HSCT.The overall survival time was not reached, the 1-year overall survival rate was 100.0%, and the 2-year overall survival rate was 90.5%. Conclusions:Auto-HSCT is a safe and effective treatment for elderly MM patients aged over 65 years after screening and assessment.