A high performance liquid chromatography (HPLC) method utilizing correction factors was established for the quantitative detection of related substances in flumazenil. Separation was achieved using an Agilent Pursuit XRs C18 column (250 mm × 4.6 mm, 5 μm) with an isocratic elution of dilute phosphoric acid, methanol, and tetrahydrofuran as the mobile phases. Correction factors calculated from a standard curve method were applied to determine the impurity content. The quantification of impurities in flumazenil was conducted using both external standard and correction factor methods, followed by validation and comparison of the two. For the identification of degradation products, a forced degradation approach was employed to prepare a flumazenil degradation solution, and the resulting impurities were confirmed by LC-MS analysis. The separation of flumazenil and its impurities was found to be efficient. The limits of quantification for impurities A, B, D, and E were established at 0.169 9, 0.314 7, 0.143 9, and 0.270 8 ng, respectively, with the limits of detection at 0.055 8, 0.096 9, 0.048 8, and 0.089 0 ng. These impurities demonstrated a strong linear relationship across the concentration ranges of 0.034 9-7.847 0, 0.038 7-8.710 7, 0.034 6-7.794 1, and 0.032 4-7.292 8 µg·mL^-1, respectively (n = 7). The method achieved average recoveries between 98.25% and 99.42%, with an RSD of less than 2.0% (n = 9), indicating high accuracy. The external standard and correction factor methods were used to determine the related substances in flumazenil, and the results of the two methods were consistent. The established HPLC method is characterized by its high accuracy, sensitivity, and repeatability, and is suitable for determining related substances in flumazenil.

OBJECTIVE: To investigate the role of multiple serological methods in the identification of complex antibodies. METHODS: The blood group antigens were detected by saline and microcolumn agglutination methods. The saline method was used to screen and identify IgM-type antibodies in the patient's serum, while the polybrene, anti-globulin, microcolumn agglutination, enzymic and absorption-elution methods were used to screen and identify IgG-type antibodies. RESULTS: The patient was B/CCDee/Jk(a-b+)/Fy(a-b+) blood type. The serum reacted with panel cells, and the reaction presented anti-E pattern in the saline medium. It was fully positive in the microcolumn agglutination card, except 2 negative ones after using papain to treat the panel cells. Referring to the pattern table, it was concluded that there existed anti-c, anti-E, and anti-Jk^a antibodies, and one antibody corresponding to an antigen that was easily destroyed by papain. The red blood cells with specific phenotype were selected for absorption-elution to identify IgG-type anti-c, anti-E, anti-Jk^a and anti-Fy^a antibodies. CONCLUSION It is confirmed that IgM-type anti-E, and IgG-type anti-c, anti-E, anti-Jk^a and anti-Fy^a antibodies exist in the patient's serum by multiple serological methods.
Humans ; Papain ; Blood Group Antigens ; Erythrocytes ; Immunoglobulin G ; Immunoglobulin M

OBJECTIVES: To investigate the clinical treatment outcomes and the changes of the outcomes over time in extremely preterm twins in Guangdong Province, China. METHODS: A retrospective analysis was performed for 269 pairs of extremely preterm twins with a gestational age of <28 weeks who were admitted to the department of neonatology in 26 grade A tertiary hospitals in Guangdong Province from January 2008 to December 2017. According to the admission time, they were divided into two groups: 2008-2012 and 2013-2017. Besides, each pair of twins was divided into the heavier infant and the lighter infant subgroups according to birth weight. The perinatal data of mothers and hospitalization data of neonates were collected. The survival rate of twins and the incidence rate of complications were compared between the 2008-2012 and 2013-2017 groups. RESULTS: Compared with the 2008-2012 group, the 2013-2017 group (both the heavier infant and lighter infant subgroups) had lower incidence rates of severe asphyxia and smaller head circumference at birth (P<0.05). The mortality rates of both of the twins, the heavier infant of the twins, and the lighter infant of the twins were lower in the 2013-2017 group compared with the 2008-2012 group (P<0.05). Compared with the 2008-2012 group, the 2013-2017 group (both the heavier infant and lighter infant subgroups) had lower incidence rates of pulmonary hemorrhage, patent ductus arteriosus (PDA), periventricular-intraventricular hemorrhage (P-IVH), and neonatal respiratory distress syndrome (NRDS) and a higher incidence rate of bronchopulmonary dysplasia (P<0.05). CONCLUSIONS There is a significant increase in the survival rate over time in extremely preterm twins with a gestational age of <28 weeks in the 26 grade A tertiary hospitals in Guangdong Province. The incidences of severe asphyxia, pulmonary hemorrhage, PDA, P-IVH, and NRDS decrease in both the heavier and lighter infants of the twins, but the incidence of bronchopulmonary dysplasia increases. With the improvement of diagnosis and treatment, the multidisciplinary collaboration between different fields of fetal medicine including prenatal diagnosis, obstetrics, and neonatology is needed in the future to jointly develop management strategies for twin pregnancy.
Bronchopulmonary Dysplasia/epidemiology* ; Female ; Gestational Age ; Humans ; Infant ; Infant, Extremely Premature ; Infant, Newborn ; Pregnancy ; Respiratory Distress Syndrome, Newborn/epidemiology* ; Retrospective Studies ; Treatment Outcome

Objective: Several COVID-19 patients have overlapping comorbidities. The independent role of each component contributing to the risk of COVID-19 is unknown, and how some non-cardiometabolic comorbidities affect the risk of COVID-19 remains unclear. Methods: A retrospective follow-up design was adopted. A total of 1,160 laboratory-confirmed patients were enrolled from nine provinces in China. Data on comorbidities were obtained from the patients' medical records. Multivariable logistic regression models were used to estimate the odds ratio ( Results: Overall, 158 (13.6%) patients were diagnosed with severe illness and 32 (2.7%) had unfavorable outcomes. Hypertension (2.87, 1.30-6.32), type 2 diabetes (T2DM) (3.57, 2.32-5.49), cardiovascular disease (CVD) (3.78, 1.81-7.89), fatty liver disease (7.53, 1.96-28.96), hyperlipidemia (2.15, 1.26-3.67), other lung diseases (6.00, 3.01-11.96), and electrolyte imbalance (10.40, 3.00-26.10) were independently linked to increased odds of being severely ill. T2DM (6.07, 2.89-12.75), CVD (8.47, 6.03-11.89), and electrolyte imbalance (19.44, 11.47-32.96) were also strong predictors of unfavorable outcomes. Women with comorbidities were more likely to have severe disease on admission (5.46, 3.25-9.19), while men with comorbidities were more likely to have unfavorable treatment outcomes (6.58, 1.46-29.64) within two weeks. Conclusion Besides hypertension, diabetes, and CVD, fatty liver disease, hyperlipidemia, other lung diseases, and electrolyte imbalance were independent risk factors for COVID-19 severity and poor treatment outcome. Women with comorbidities were more likely to have severe disease, while men with comorbidities were more likely to have unfavorable treatment outcomes.
Adult ; Aged ; COVID-19/virology* ; China/epidemiology* ; Comorbidity ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Severity of Illness Index ; Treatment Outcome

AIM:To investigate the effect of histone deacetylase 1(HDAC1)silencing on apoptosis of squa-mous cell carcinoma of skin.METHODS:Skin squamous cell carcinoma A431 cells were transfected with HDAC1 small interfering RNA(HDAC1 siRNA)or small interfering RNA negative control(siRNA NC).The expression levels of HDAC1 in transfected cells were detected by RT-PCR and Western blot.The cell viability was measured by MTT assay, and the apoptosis was analyzed by flow cytometry.The protein levels of STAT3,p-STAT3 and cleaved caspase-3 were de-termined by Western blot.The inhibitor of STAT3 signaling pathway was used to treat the A 431 cells transfected with HDAC1 siRNA.The cell viability was detected by MTT assay,the apoptosis was analyzed by flow cytometry,and the pro-tein levels of STAT3,p-STAT3 and cleaved caspase-3 were determined by Western blot.RESULTS: HDAC1 siRNA in-hibited the expression of HDAC1 at mRNA and protein levels in the A431 cells.After interfering with the expression of HDAC1,the cell viability and the protein level of p-STAT3 in the cells decreased,while the apoptotic rate and the protein level of cleaved caspase-3 in the cells were increased.After treatment with the inhibitor of STAT3 pathway,the viability of A431 cells transfected with siRNA and the protein level of p-STAT3 decreased,while the apoptotic rate and the protein le-vel of cleaved caspase-3 in the cells were increased.CONCLUSION: Interference with HDAC1 expression may regulate the STAT3 signaling pathway to inhibit the viability of skin squamous cell carcinoma cells,thus promoting the apoptosis of squamous cell carcinoma of skin.

Ca²⁺/calmodulin-dependent protein kinase II (CaMKII) is the most abundant kinase within excitatory synapses in the mammalian brain. It interacts with and phosphorylates a large number of synaptic proteins, including major ionotropic glutamate receptors (iGluRs) and group I metabotropic glutamate receptors (mGluRs), to constitutively and/or activity-dependently regulate trafficking, subsynaptic localization, and function of the receptors. Among iGluRs, the N-methyl-D-aspartate receptor (NMDAR) is a direct target of CaMKII. By directly binding to an intracellular C-terminal (CT) region of NMDAR GluN2B subunits, CaMKII phosphorylates a serine residue (S1303) in the GluN2B CT. CaMKII also phosphorylates a serine site (S831) in the CT of α-amino-3-hydroxy-5- methylisoxazole-4-propionic acid receptors. This phosphorylation enhances channel conductance and is critical for synaptic plasticity. In addition to iGluRs, CaMKII binds to the proximal CT region of mGluR1a, which enables the kinase to phosphorylate threonine 871. Agonist stimulation of mGluR1a triggers a CaMKII-mediated negative feedback to facilitate endocytosis and desensitization of the receptor. CaMKII also binds to the mGluR5 CT. This binding seems to anchor and accumulate inactive CaMKII at synaptic sites. Active CaMKII dissociates from mGluR5 and may then bind to adjacent GluN2B to mediate the mGluR5-NMDAR coupling. Together, glutamate receptors serve as direct substrates of CaMKII. By phosphorylating these receptors, CaMKII plays a central role in controlling the number and activity of the modified receptors and determining the strength of excitatory synaptic transmission.
Calcium-Calmodulin-Dependent Protein Kinase Type 2 ; metabolism ; Neuronal Plasticity ; Phosphorylation ; Receptor, Metabotropic Glutamate 5 ; metabolism ; Receptors, Metabotropic Glutamate ; metabolism ; Receptors, N-Methyl-D-Aspartate ; metabolism ; Serine ; metabolism ; Synapses ; Synaptic Transmission

OBJECTIVETo observe the electroacupuncture (EA) pretreatment at Baihui (GV20) on the concentration of adenosine deaminase (ADA) and adenosine, and to evaluate its effects on the neurologic function score and the infarction volume after middle cerebral artery occlusion (MCAO) ischemia/reperfusion (I/R), thus exploring its mechanisms for relieving the ischemia/reperfusion injury.

METHODSTotally 54 male SD rats were randomly divided into 3 groups, the sham-EA group, the EA group, and the control group, 18 in each group. Rats in the control group were not intervened after anesthesia. Rats in the EA group were needled at Baihui (GV20) for 30 min. Rats in the sham-EA group received the same procedure as those performed in the EA group without electricity connected. The changes of adenosine and ADA contents were detected at 30, 60, and 120 min after EA respectively. The I/R model was established. Totally 48 male SD rats were randomly divided into 6 groups, i.e., the model group (Group A), the EA group (Group B), the EA +8-Cyclopentyl-1,3-dipropylxanthine (DPCPX) group (Group C), the EA + DMSO group (Group D), the Deoxycoformycin (Deo) group (Group E), and the normal saline group (Group F). Rats in Group B, C, and D received EA for 30 min before modeling. Rats in Group C and D were peritoneally injected with DPCPX (1 mg/kg) and DMSO (1 mL/kg) at 30 min before EA. The neurologic function score was evaluated and the infarct volumes were detected after 24-h reperfusion.

RESULTSCompared with the sham-EA group, there was no statistical difference in the contents of the adenosine or ADA in the control group at each time point (P > 0.05). Compared with the control group at the same time point, the content of ADA significantly decreased at 60 min in the EA group [(315.0 +/- 22.9 U/L), P < 0.05], and restored to the normal level at 120 min after EA. The content of adenosine increased in the EA group at 120 min [(20.4 +/- 2.2) ng/microL, P < 0.05]. Compared with the model group, the neurologic function score decreased (P < 0.05) and the infarct volumes were obviously reduced (P < 0.01) in Group B, D and E. There was no statistical difference in the neurologic function score or the infarct volumes in other groups, when compared with the model group (P > 0.05)

CONCLUSIONEA at Baihui (GV20) showed protective effects on the cerebral I/R rats, which might be achieved through lowering the ADA concentration and elevating the adenosine content, and further activating adenosine A1 receptor.

Adenosine Deaminase ; metabolism ; Animals ; Brain Ischemia ; metabolism ; Electroacupuncture ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism

INTRODUCTIONCirculating insulin concentrations provide important information for the evaluation of insulin secretion and insulin resistance. Reference intervals are the most widely applied tool for the interpretation of clinical laboratory results. We carried out an analysis of the data available from the Fangchenggang Area Male Health and Examination Survey in order to derive a reference interval for fasting insulin specific to the Chinese population.

METHODSA total of 1,434 fasting serum insulin results were obtained from healthy nondiabetic adult men aged 20-69 years, after taking into consideration the inclusion and exclusion criteria. Serum insulin was measured using electrochemiluminescence immunoassays. Nonparametric statistical methods were used to calculate and analyse the data.

RESULTSThe reference interval for fasting serum insulin for Chinese adults was in the range 1.57-16.32 μU/mL (median 5.79 μU/mL). Significant correlations were found between fasting serum insulin and glucose and diastolic blood pressure (p < 0.001). Statistically significant differences were observed in insulin concentration with respect to age and body mass index (BMI; p < 0.001). Younger people had a higher fasting serum insulin concentration. Increased fasting serum insulin was also found to be associated with BMI.

CONCLUSIONWe established a reference interval for fasting serum insulin in healthy nondiabetic adult Chinese men that is lower than what was previously suggested. BMI and age (but not smoking, alcohol consumption or physical activity) were found to be important factors associated with fasting serum insulin. Our results will help improve the diagnostic interpretation of investigations for metabolic and cardiovascular disorders in a Chinese population.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Body Mass Index ; China ; Fasting ; blood ; Humans ; Insulin ; blood ; Insulin Resistance ; physiology ; Male ; Middle Aged ; Nomograms ; Reference Values ; Retrospective Studies ; Young Adult

Objective To evaluate the clinical outcome of transesophageal echocardiography(TEE) in guiding and monitoring min-invasive transthoracic hybrid procedure for intracristal ventricular septal defects (VSD) using eccentric occluders.Methods 13 patients with intacristal VSD successfully underwent the min-invasive hybrid procedure using eccentric occluders were analyzed retrospectively.They were all carefully examined by transthoracic echocardiography (TTE) preoperatively and were diagnosed as intracristal VSD.There were no obvious aortic valve prolapse and no severe regurgitation.The entire process was guided and monitored by TEE.The condition of VSD occlusion was assessed by immediate postoperative TEE.Results All cases were occluded successfully.The results of postoperative TEE showed that the occluder was coincided closely with the edge of VSD,no aortic valve regurgitation worsen.2 had tiny residual left-to-right shunt (＜1.5 mm),which disappeared 1 and 3 months respectively after operation.No occluder dislocated followed-up at 6 months to 2 years postoperatively,and no major complications occurred.Conclusions The min-invasive hybrid procedure for intracristal VSD using eccentric occluders guided and monitored by TEE is easy to manipulate,which is safe,less invasive,and has excellent early-term results.

The present study aimed to investigate the neuro-cognitive features in the processing of vocabularies of date in Chinese, using block-design functional magnetic resonance imaging (fMRI). Nineteen normal right-handed volunteers whose native language was Chinese performed judgments of vocabulary of month (JVM), the orientation of digit (JOD) and the meaning of words (JMW) respectively, while the fMRI data were recorded by Signa HDe 1.5T MR machine. All design of three tasks was adapted from previous studies with slight modification. The JOD and JWM were investigated as contrast conditions. JVM asked the subjects to determine whether the month belonged to the first half of the year. JOD was to tell whether the third digit had the same orientation (upright orientation or italic orientation) as the first two, and JMW was to determine whether the two-Chinese-character word was animate. The subjects responded according to the task instruction with a button pressing. Statistical parametric mapping (SPM2) was employed to process data and localize functional areas. We compared the average activation intensity of each activated brain regions in the same task against the rest and the activation intensity of the same regions in different tasks respectively. The activations in inferior parietal (BA40) and inferior occipital (BA18/19) were found in the JVM and JOD. The same areas in middle frontal (BA6), fusiform (BA18), posterior in right cerebellum areas were activated during the JVM and JMW. When the activation of the JMW was subtracted from the JVM, many areas concerning numerical processing, such as left anterior cingulate (BA32), post central (BA2), and the right superior temporal (BA39), superior parietal (BA7), as well as the inferior parietal (BA40), precuneus (BA7/19) of the bilateral hemispheres, were significantly activated. When the activation of the JOD was subtracted from the JVM, the left inferior occipital and the fusiform (BA18) of the bilateral hemispheres, which were also involved in the linguistic processing according to previous studies, were significantly activated. The above results prove that the processing of vocabulary of months in Chinese involves not only linguistic processing but also numerical processing; and these results further indicate that Chinese people might gain access to the cognitive experience of number during the process of acquisition of native language.
Brain ; anatomy & histology ; Brain Mapping ; China ; Humans ; Magnetic Resonance Imaging ; Vocabulary