Rational design of a near-infrared dual-emission fluorescent probe for ratiometric imaging of glutathione in cells.

Sensors for which the output signal is an intensity change for a single-emission peak are easily disturbed by many factors, such as the stability of the instrument, intensity of the excitation light, and biological background. However, for ratiometric fluorescence sensors, the output signal is a change in the intensity ratio of two or more emission peaks. The fluorescence intensity of these emission peaks is similarly affected by external factors; thus, these sensors have the ability to self-correct, which can greatly improve the accuracy and reliability of the detection results. To accurately image glutathione (GSH) in cells, gold nanoclusters (AuNCs) with intrinsic double emission at wavelengths of 606 nm and 794 nm were synthesized from chloroauric acid. With the emission peak at 606 nm as the recognition signal and the emission peak at 794 nm as the reference signal, a near-infrared dual-emission ratio fluorescence sensing platform was constructed to accurately detect changes in the GSH concentration in cells. In vitro and in vivo analyses showed that the ratiometric fluorescent probe specifically detects GSH and enables ultrasensitive imaging, providing a new platform for the accurate detection of active small molecules.

Anesthesia management experience for pediatric day-case PDA ligation under thoracoscopy assisted by a robot: a retrospective study.

BACKGROUND: To summarize the anesthesia management experience for pediatric day-case patent ductus arteriosus (PDA) ligation under robot-assisted thoracoscopy and explore the key points of anesthesia management for this procedure. METHODS: The clinical data of 72 pediatric patients who underwent robot-assisted thoracoscopic day-case PDA ligation at the Children's Hospital, Zhejiang University School of Medicine from April 2021 to February 2023 were retrospectively analyzed. 0.3% ropivacaine local infiltration combined with S-ketamine 0.2 mg/kg intravenous injection was used for postoperative analgesia The patient's basic information and intraoperative conditions were analyzed, which included gender, age, weight, surgery time, anesthesia time, extubation time, intraoperative blood loss, MAP before pneumothorax, PaCO2 before pneumothorax, etc. Postoperative conditions were also monitored, such as PACU stay time, agitation during the recovery period, pain, and the incidence of nausea and vomiting. After discharge, the recovery status was assessed. RESULTS: A total of 70 pediatric patients who met the criteria for day-case PDA ligation were included in this study. Before the occurrence of pneumothorax, the mean arterial pressure (MAP) of these 70 patients was 69.58 ± 12.52 mmHg, and during controlled hypotension, the MAP was 54.96 ± 11.23 mmHg. Before the occurrence of pneumothorax, the partial pressure of carbon dioxide (PaCO2) was 38.69 ± 3.38 mmHg, and during controlled hypotension, the PaCO2 was 51.42 ± 4.05 mmHg. Three cases experienced agitation during the recovery period, and four cases had mild pain, but there was no moderate or severe pain, nausea, or vomiting. Only 1 case of postoperative respiratory tract infection and 1 case of postoperative pneumothorax occurred. Within 30 days after discharge, the unplanned revisit rate, unplanned readmission rate, and surgical wound infection rate were all 0. The residual shunt rate detected by echocardiography was 0 after 1 month. CONCLUSIONS: The children under the robot-assisted thoracoscopic day case PDA surgeries in this study have limited trauma, little bleeding, and little postoperative pain, though still at a risk of respiratory infection and pneumothorax.

Application of laryngeal mask airway anesthesia with preserved spontaneous breathing in children undergoing video-assisted thoracic surgery.

Purpose: To investigate the feasibility and safety of non-intubated general anesthesia with spontaneous breathing combined with paravertebral nerve blocks (PVNB) in young children undergoing video-assisted thoracic surgery (VATS) and to determine its significance for rapid recovery after pediatric thoracic surgery. Methods: The data of 46 children aged 6-36 months with an American Society of Anesthesiologists status of I-II who underwent elective VATS under general anesthesia were retrospectively analyzed. Of these patients, 25 underwent non-intubated general anesthesia with spontaneous breathing combined with PVNB (non-intubation group), and 21 received conventional intubated general anesthesia combined with local infiltration anesthesia (intubation group). The following perioperative parameters were compared between the two groups: heart rate (HR), mean arterial pressure, saturation of pulse oximetry (SpO2), partial pressure end-tidal carbon dioxide, time from the completion of the operation to extubation or removing laryngeal masks, time to first feeding after the operation, length of postoperative in-hospital stay, incidence of postoperative complications, and hospitalization expenses. Results: The operations were completed successfully in both groups. When the non-intubation group was compared with the intubation group, the minimal SpO2 level during the surgery was higher (93% vs. 88%, P < 0.001), which might indicate better oxygenation. There was no significant difference of the duration of surgery and intraoperative blood loss between two groups. Compared to the intubation group, the duration of anesthesia (P = 0.027), time from the completion of the operation to extubation (P < 0.001), time to the first feeding after surgery (P < 0.001), and length of postoperative in-hospital stay (P < 0.001) were significantly reduced in the non-intubation group. The incidence of postoperative complications was not significantly different. Conclusions: Non-intubated general anesthesia with spontaneous breathing combined with PVNB is safe and feasible in young children undergoing VATS and can promote rapid recovery in young children undergoing thoracoscopic surgery.

Gut-Derived Exosomes Induce Liver Injury After Intestinal Ischemia/Reperfusion by Promoting Hepatic Macrophage Polarization.

Liver injury induced by intestinal ischemia/reperfusion (I/R) is accompanied by the polarization of Kupffer cells, which are specialized macrophages located in the liver. However, the causes of hepatic macrophage polarization after intestinal I/R remain unknown. This study investigated whether gut-derived exosomes contribute to the pathogenesis of liver injury triggered by intestinal I/R in a murine model and explored the underlying mechanisms. Intestinal I/R models were established by temporally clamping the superior mesenteric arteries of mice. Exosomes were isolated from the intestinal tissue of mice that underwent intestinal I/R or sham surgery according to a centrifugation-based protocol. Exosomes were co-cultured with RAW 264.7 macrophages or injected intravenously in mice. Liposomal clodronate was administered intraperitoneally to deplete the macrophages. Macrophage polarization was determined by flow cytometry, immunohistochemistry, and quantitative polymerase chain reaction. Liver injury was assessed by histological morphology and increased serum aspartate aminotransferase and alanine aminotransferase levels. Exosomes from mice intestines subjected to I/R (IR-Exo) promoted macrophage activation in vitro. Intravenous injection of IR-Exo caused hepatic M1 macrophage polarization and led to liver injury in mice. Depleting macrophages ameliorated liver injury caused by intestinal I/R or the injection of IR-Exo. Furthermore, inhibiting exosome release improved intestinal injury, liver function, and survival rates of mice subjected to intestinal I/R. Our study provides evidence that gut-derived exosomes induce liver injury after intestinal I/R by promoting hepatic M1 macrophage polarization. Inhibition of exosome secretion could be a therapeutic target for preventing hepatic impairment after intestinal I/R.

The Characteristics and Distribution of Nerve Plexuses in the Dartos Fascia From Concealed Penis Children.

The purpose of this study is to analyze the nerve plexus distribution in dartos fascia of concealed penis (CP). A total of 28 CP patients met ASA categories I and II were included, with median age of 3.5 years (8 months-5 years). During the surgery, tissue samples of dartos fascia at points 3, 6, 9, and 12 o'clock of the penile shaft were collected. Standard hematoxylin and eosin (H&E) staining and S-100 immunohistochemical staining were used to analyze the nerve plexus distribution among different positions. The number of nerve plexuses in superficial fascia collected at the 6 o'clock position of the penile shaft was the most abundant among four positions (median 7.25, range 1-24). The abundant nerve plexuses in the dartos fascia of CP patients, especially at the 6 o'clock position, indicate that the surgery on the preputial frenulum should avoid damage to the dartos fascia, as it might be related to maintain the erection and sexual function in adolescence.

Exploring the mechanism of PingTang No.5 capsule on nonalcoholic fatty liver disease through network pharmacology and experimental validation.

PingTang No.5 capsule (PT5), a modified Traditional Chinese Medicine (TCM) formula of Zexie Decoction, is used to treat patients with lipid metabolism disorders in our hospital. The present study was designed to investigate the mechanisms of PT5 in treating non-alcoholic fatty liver disease (NAFLD). PT5 information including ingredients, pharmacological properties, and potential targets was obtained from TCM databases. The candidate targets of PT5 were predicted by network pharmacological analysis, and the possible pathway and mechanism were obtained from DAVID database, followed by experimental validation in NAFLD mice model treated with PT5. Total 328 compounds were selected using the threshold oral bioactivity (OB) > 30% or drug-likeness (DL) > 0.1 of pharmacology characteristic, and 1033 candidate targets obtained to construct the network analysis. The 113 targets were selected from the intersection between candidate targets of PT5 and NAFLD relative gene. These targets were evaluated in diabetic complications, cancer, Hepatitis B, Fluid shear stress and atherosclerosis, and TNF signaling pathway. TNF-α was the important factor in protein interaction analysis of STRING and involved in the lipid regulation and oxidative stress in NAFLD. When administrated to the NAFLD mice, PT5 reduced weight, blood fatty acids, decreased the adipocyte size, and improved the metabolism. Besides, the molecular verification of lipid metabolism increased and oxidative stress reduced that interpreted the mechanism of PT5 preventing liver cell from lipid accumulation and injury of NAFLD. These results presented PT5 have the potential therapy as an alternative treatment for NAFLD.

Effect of nalbuphine on rocuronium injection pain.

Objective: Rocuronium-associated intravenous injection pain occurs frequently in children during induction of anesthesia. The aim of this study was to systematically evaluate the benefits of nalbuphine in patients with rocuronium-associated injection pain. Methods: Ninety children undergoing tonsillectomy and adenoidectomy in our hospital between October 2019 and September 2020 were randomly divided into the following groups, with 30 patients per group: control group (group C), lidocaine group (group L), and nalbuphine group (group N). Routine 0.1 mg/kg midazolam and 2 mg/kg propofol were injected intravenously. After sedation, children in group C, group L, and group N were administered an intravenous injection of saline, lidocaine (10 mg/mL), or nalbuphine hydrochloride (2 mg/mL), respectively, at a dosage of 0.1 mL/kg. Intravenous injection of rocuronium stock solution (0.6 mg/kg) was administered 2 minutes later. Pain was evaluated using Ambeshs 4-pointscale. The incidence of rocuronium injection pain was compared among the three groups, and postoperative adverse reactions, such as drowsiness, bradycardia, hypotension, and respiratory depression, were evaluated. Results: The incidence of injection pain among children in group N was significantly lower than that in group C and group L (p<0.05). The incidence of drowsiness in group N was significantly higher than that in the other groups (p<0.05). The incidences of hypotension, bradycardia, and respiratory depression were not significantly different among the three groups (p>0.05). Conclusions: Intravenous nalbuphine during induction of anesthesia effectively prevented rocuronium-associated injection pain in children. Drowsiness is a complication.

Recommendation about the perioperative prevention of infection to healthcare workers and the anesthesia management of children with SARS-CoV-2 infection.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has spread widely and persistently over 100 countries. New challenges have occurred in the perioperative management of airway and anesthesia in children diagnosed with SARS-CoV-2 infection. According to current publications and to our own experiences in anesthesia management for cases with SARS-CoV-2 suspected, we reviewed concerns about the perioperative prevention of SARS-CoV-2 to medical staff and the anesthesia strategy to the patient.

Application of end-tidal carbon dioxide monitoring via distal gas samples in ventilated neonates.

BACKGROUND: Previous research has suggested correlations between the end-tidal partial pressure of carbon dioxide (PETCO2) and the partial pressure of arterial carbon dioxide (PaCO2) in mechanically ventilated patients, but both the relationship between PETCO2 and PaCO2 and whether PETCO2 accurately reflects PaCO2 in neonates and infants are still controversial. This study evaluated remote sampling of PETCO2 via an epidural catheter within an endotracheal tube to determine the procedure's clinical safety and efficacy in the perioperative management of neonates. METHODS: Abdominal surgery was performed under general anesthesia in 86 full-term newborns (age 1-30 days, weight 2.55-4.0 kg, American Society of Anesthesiologists class I or II). The infants were divided into 2 groups (n = 43 each), and carbon dioxide (CO2) gas samples were collected either from the conventional position (the proximal end) or a modified position (the distal end) of the epidural catheter. RESULTS: The PETCO2 measured with the new method was significantly higher than that measured with the traditional method, and the difference between PETCO2 and PaCO2 was also reduced. The accuracy of PETCO2 measured increased from 78.7% to 91.5% when the modified sampling method was used. The moderate correlation between PETCO2 and PaCO2 by traditional measurement was 0.596, which significantly increased to 0.960 in the modified sampling group. Thus, the PETCO2 value was closer to that of PaCO2. CONCLUSION: PETCO2 detected via modified carbon dioxide monitoring had a better accuracy and correlation with PaCO2 in neonates.

Effects of Heart Bypass Surgery on Plasma Aß40 and Aß42 Levels in Infants and Young Children.

Accumulation of ß-amyloid (Aß) plaques is a pathological hallmark of Alzheimer disease. Aß levels in animals and adults were reported to be associated with postoperative cognitive dysfunction (POCD). Our goal was to determine the plasma levels of Aß in infants and young children after cardiac surgery with cardiopulmonary bypass (CPB).Forty-two infants and young children aged from 1 to 35 months undergoing cardiac surgery with general anesthetics were prospectively enrolled from January to June 2014 at a tertiary medical center. Perioperative plasma samples were obtained, and Aß42 and Aß40 levels were measured using ELISA. Other clinical characteristics of the patients were also recorded.Plasma levels of Aß42 and Aß40 decreased dramatically 2 hours after surgery and remained significantly lower 6 hours after operation. Baseline Aß42 level correlated significantly with surgical intensive care unit (SICU) length of stay (LOS) and was an independent predictor for SICU LOS on multivariate analysis.Cardiac surgery with CPB decreases plasma Aß levels. Plasma levels of Aß42 and Aß40 might be used as novel biomarkers for predicting outcomes in the patient population.

Effects of traditional Chinese medicine Shuxuetong injection on random skin flap survival in rats.

BACKGROUND: A Shuxuetong injection is traditionally used in Chinese medicine to treat "blood stasis and stagnation" (yu xue yu zhi). We investigated the effect of such injection on the survival of random skin flaps. METHODS: McFarlane flaps were established in 60 rats divided into two groups. Postoperative celiac injections were given to both groups for 7 days. Shuxuetong was injected into the test group, and saline was injected into controls. On day 7, tissues were stained with H&E (hematoxylin-eosin) stain, immunohistochemically evaluated, and the expression levels of xanthine oxidase were determined. RESULT: The mean area of flap survival in the test group was significantly higher than in controls. Expression of vascular endothelial growth factor and superoxide dismutase, and microvessel development, were markedly increased in the test group, and the malondialdehyde level was reduced. CONCLUSION: Shuxuetong promotes random skin flap survival.

Mechanism of simvastatin-induced K562 cell apoptosis.

Statins are being widely used for the therapy and prevention of several types of tumors, including human chronic myelogenous leukemia, but the underlying molecular mechanisms still remain unknown. Therefore, inhibition of cell proliferation, apoptosis and involved molecules were investigated in K562 cells after incubation with simvastatin.The results showed that simvastatin diminished K562 cell proliferation and induced apoptosis. At the same time, the level of reactive oxygen species (ROS) and intracellular calcium concentration increased. Furthermore, nitric oxide (NO) content and inducible NO synthase (iNOS) mRNA expression were significantly higher in the simvastatin-treated group than in the corresponding control group. The elevated ROS level and intracellular calcium concentration, enhanced mRNA expression of iNOS and total NO content might be responsible for the apoptotic and anti-proliferative effects of simvastatin in K562 cells.

In vitro and in vivo study of cell growth inhibition of simvastatin on chronic myelogenous leukemia cells.

BACKGROUND: Statins, a family of 3-hydroxy-3-methylglutaryl CoA (HMG CoA) reductase inhibitors, are being investigated for the therapy and prevention of cancers. Here we aimed to investigate the effects of simvastatin on chronic myelogenous leukemia (CML) cells in vitro and in vivo, and to elucidate the mechanisms. METHODS: Cell proliferation and cell cycle were measured after K562 cells were incubated with simvastatin, and differentially expressed genes were determined by oligonucleotide microarray. Changes of 2 genes obtained by oligonucleotide microarray were validated by real-time RT-PCR, and immunohistochemistry was performed to determine expression of proliferating cell nuclear antigen (PCNA). Finally, a xenograft tumor model was constructed to evaluate the effects of simvastatin in vivo. RESULTS: Simvastatin could inhibit K562 cell proliferation, and the inhibition rate was approximately 30% after treatment with 20 mumol/l simvastatin for 48 h. Cell cycle was arrested in G(1) phase, as shown by flow cytometry results. Fifteen downregulated, 9 upregulated cell cycle-related genes and decreased PCNA protein were observed in the presence of simvastatin. Furthermore, simvastatin exhibited impairment of xenograft tumor growth in nude mice and also blocked cell cycle in G(1) phase. CONCLUSION: Simvastatin can inhibit CML cell proliferation in vitro and in vivo, and its mechanisms might be involved in cell cycle regulation.

[Simvastatin-induced apoptosis of K562 cells is mediated by endoplasmic reticulum stress].

To explore the apoptotic effect of simvastatin on K562 cells through endoplasmic reticulum stress, morphological change of apoptotic cells was observed by Hoechst33258 fluorescent staining under fluorescent microscope. Apoptosis rate of cells was determined with annexinV-FITC/PI double staining by flow cytometry; Intracellular calcium concentration ([Ca2+]i) was measured by laser scanning confocal microscope (LSCM); The expression levels of glucose regulated protein 78 (GRP78) and calpain gene mRNA were determined by RT-PCR; The expression levels of caspase-3, -6, -7, -9, -12, calpain and GRP78 proteins were evaluated by Western blotting. In this study, K562 cells treated with simvastatin for 72 h exhibited typical morphological change of apoptosis cells. After 72 h exposed to 10, 20, 30 micromol x L(-1) simvastatin, the apoptotic rates of K562 cells were 12.41%, 19.08% and 23.41%, respectively. Simvastatin induced the increase of [Ca2+]i in K562 cells, fluorescent intensities were 43, 54, and 64, respectively. The expression levels of GRP78 and calpain gene mRNA were up-regulated. The cleavage and activation of caspase-3, -6, -7, -9, -12 and upregulation of GRP78 expression were determined by Western blotting. These findings suggest that endoplasmic reticulum is an important pathway of apoptosis in cells and participates simvastatin-induced apoptosis in K562 cells. It is implied that simvastatin may be suitable for clinical usage in the treatment of myeloma patients.

[Experimental research of targeting hTERT gene inhibited in hepatocellular carcinoma therapy by RNA interference].

BACKGROUND & OBJECTIVE: RNA interference (RNAi) is a new gene blocking technology that silences target gene at post-transcription level induced by the small interference RNA (siRNA). RNAi has been demonstrated great prospect in gene functional research and gene therapy areas. Nowadays, RNAi has been reported to be used to inhibit the expression of endogenous genes including cyclophilin, GAPDH, p53, and c-myc; and there were some progresses in the therapy of the diseases caused by AIDS and hepatitis viruses with RNAi. However, hTERT gene, which was highly expressed in hepatocellular carcinoma and other malignant neoplasm, has not been researched by RNAi. In present research, we utilized RNAi to inhibit hTERT gene expression in vitro and in vivo, investigated the feasibility and specificity of gene therapy for hepatocellular carcinoma. METHODS: Small interference RNAs homologous to hTERT gene were designed,pTZU6+1-shRNA-hTERT vector was constructed and transfected into hepatocellular carcinoma SMMC-7721 cells and transplanted SMMC-7721 tumor in nude mice to induce RNAi. The changes of hTERT gene expression and tumor cell proliferation in both siRNA treatment groups and control group were determined by flow cytometry, reverse transcription polymerase chain reaction (RT-PCR), immunochemistry in vitro and in vivo. RESULTS: The expression of hTERT had been obviously inhibited by RNAi in vitro. The inhibition rate of cell growth was 37.5% after pTZU6+1-shRNA-hTERT vector was transfected to hepatocellular carcinoma SMMC-7721 cells; the phase of cell cycle indicated the reduction of S phase, while G(1)/G(0) phase increased. The mRNA level of hTERT decreased from 99.4% to 53.1%, its protein expression reduced from 86.3% to 46.6%. The tumor size reduced after treated with pTZU6+1-shRNA-hTERT vector in vivo; hTERT mRNA level decreased from 99.1% to 76.2%, and its protein expression decreased from 87.2% to 61.8% in siRNA treatment group. In contrast, there were no changes in control groups in vitro and in vivo. CONCLUSION: RNAi inhibits the hTERT gene expression and proliferation of hepatocellular carcinoma SMMC-7721 cells with specificity, and is a possible new approach for neoplasm gene therapy.

[Detection of platelet Ca2+(i), CD62P, CD63 and plasma CD62P in cirrhosis patients].

OBJECTIVES: To study on the relationship between platelet Ca2+(i), CD62P, CD63, serum CD62P (SCD62P) and cirrhosis patients. METHODS: Platelet CD62P, CD63 were determined with flow cytometry, SCD63P with ELISA, and Ca2+(i) in platelet was determined with fluorophotometry. RESULTS: Platelet Ca2+(i), CD62P, CD63, and SCD62P levels in cirrhosis patients were (103.1+/-22.2)nmol/L, (47.6+/-20.0)%, (47.1+/-24.6)%, and (67.6+/-37.6)microg/L, and in controls were (57.6+/-13.1)nmol/L, (3.1+/-0.7)%, (2.5+/-0.7)%, and (24.0+/-6.5)microg/L, respectively. The levels in the former were higher than those in the latter (t > or = 6.148, P<0.05). The above levels in upper gastrointestinal haemorrhage group were much higher than those in the non-haemorrhage group (120.3nmol/L+/-18.8nmol/L vs 91.1nmol/L+/-14.3nmol/L, 64.9%+/-14.7% vs 34.6%+/- 11.9%, 70.9%+/-14.5% vs 30.2%+/-14.4%, and 103.6microg/L+/-14.9microg/L vs 40.8microg/L+/-24.0microg/L, respectively, t > or = 5.380, P<0.05). But the numbers of platelet between the two groups were no obvious difference. CONCLUSIONS: Platelet in the cirrhosis patients is greatly active, and the detection of platelet CD62P, CD63, SCD62P has a certain value in judging the degree of cirrhosis.