[Objective] To investigate the factors influencing the detection of HLA-C expression by flow cytometry. [Methods] A total of 12 hematopoietic stem cell suspension samples from peripheral hematopoietic stem cell volunteer donors were randomly collected after CD34⁺ cell counting detection. The influence of detecting different number of nucleated cell (500 000, 50 000 and 5 000), sequential order of red blood cell lysis and antibody incubation, and the HLA-C antibody with varied remaining time from the expiration date on the detection results of HLA-C expression by flow cytometry were investigated, respectively. The significance of differences between different groups was analyzed through Student t test. [Results] There was no significant difference in the proportion of HLA-C positive cells and mean fluorescence intensity (MFI) among the three groups with different nucleated cell numbers detected (500 000, 50 000 and 5 000) (P>0.05). The sequential order of red blood cell lysis and antibody incubation had no influence on the proportion of HLA-C positive cells (P>0.05), but HLA-C MFI value was significantly lower when antibody incubation was performed after red blood cell lysis than that when antibody incubation was performed before red blood cell lysis (P<0.05). The proportion of HLA-C positive cells and MFI value detected by HLA-C antibody remaining 24 months from the expiration date were significantly higher than those detected by HLA-C antibody remaining only 5 months from the expiration date (P<0.05). [Conclusion] The present study has investigated the factors of influencing HLA-C expression level by flow cytometry, the results have important reference and application value for standardizing the experimental operation of HLA-C expression and improving the accuracy and comparability of detection results.

BackgroundSleep disorder in the first trimester is a fairly common health problem， and previous studies have mainly reflected the overall sleep quality through scale assessments， which may not accurately capture the differences among various subtypes. ObjectiveTo explore the latent profiles of sleep quality in first-trimester pregnant women and identify the physiological， psychological and social factors， in order to provide practical references for the development of personalized interventions for sleep disorders in first-trimester pregnant women. MethodsA total of 1 066 first-trimester pregnant women who visited the obstetric outpatient clinic of a tertiary A hospital in Shenzhen from October 2021 to October 2022 were investigated using the general information questionnaire， Pittsburgh Sleep Quality Index （PSQI）， Edinburgh Postnatal Depression Scale （EPDS）， Chinese version of short International Physical Activity Questionnaire （IPAQ-S-C） and Social Capital Assessment Tool in Pregnancy for Maternal Health （SCAT-MH）. Then the sleep profiles were identified through latent profile analysis， and the robust mixture regression model was employed to determine the influencing factors of sleep profiles. ResultsA 3-profile solution showed the best fit： 732 cases （68.67%） of good sleep quality group， 87 cases （8.16%） of low sleep efficiency group， and 247 cases （23.17%） of daytime dysfunction group. In comparison with subjects in good sleep quality group， first-trimester pregnant women in low sleep efficiency group were at a younger age （OR=0.951， 95% CI： 0.922~0.980）， held a Bachelor's degree or above （OR=1.869， 95% CI： 1.260~2.773） and exhibited lower levels of social capital （OR=0.962， 95% CI： 0.951~0.973）， while those in daytime dysfunction group were at an older age （OR=1.072， 95% CI： 1.027~1.120） and had higher levels of depression （OR=1.166， 95% CI： 1.115~1.218）. Pregnant women who were workers （OR=0.507， 95% CI： 0.293~0.876） were less likely to report daytime dysfunction. ConclusionThree latent profiles with significant heterogeneity are derived from the sleep quality of first-trimester pregnant women， and levels of depression and social capital are the main influencing factors of sleep quality. ［Funded by Industry-University-Research Innovation Fund for Chinese Universities （number， 2023HT018）］

Objective To analyze and summarize the clinical presentation of spontaneous and secondary intracranial hypotension syndrome(IHS).Methods Patients diagnosed with spontaneous or secondary IHS from September 2022 to May 2023 were retrospectively analyzed.The clinical data,imaging features,treatment methods and prognosis were collected.The correlation between intracranial pressure values and clinical characteristics of the patients was statistically analyzed.Results A total of 30 patients were enrolled,and the proportion of spontaneous and secondary IHS was 63%(19 cases)and 37%(11 cases),respectively.In terms of clinical features,orthostatic headache was the most common type(29 cases,96.7%)and most commonly involved occipital region(12 cases,40.0%),followed by frontoparietal region(9 cases,30.0%).Among the brain imaging features,dural enhancement was the most common(17 cases,56.7%).According to CT angiography of spinal cord findings,cerebrospinal fluid leakage is one of the most common location of cervical spine segments(10 cases),and on the thoracic segments(9 cases),followed by the thoracic segments(4 cases)and lumbar segments(4 cases).After conservative treatment and surgical treatment,the total effective rate was 90%.Conclusion Orthostatic headache and cranial MRI"dural enhancement"have strong indication on the definitive diagnosis of IHS.CT myelography is helpful to precisely localize the site of cerebrospinal fluid leakage.Targeted epidural blood patch therapy is an effective method to cure IHS when conservative treatment is ineffective.

Objective To investigate the correlation of serum osteoprotegerin (OPG) and calcium levels with cerebral microbleeds in patients with acute ischemic stroke. Methods A total of 97 patients with acute ischemic stroke were selected as the study subjects and divided into cerebral microbleed (group 31 patients with) and non-cerebral microbleed (group 66 patients) based on the results of susceptibility-weighted imaging. Demographic data and laboratory examination indicators were collected from the two groups, and serum OPG and calcium levels were measured. The levels of serum OPG and calcium were compared between patients with different degrees of lesion and bleeding sites. Spearman rank correlation analysis was used to determine the correlations of serum OPG and calcium with cerebral microbleeds. Multivariate Logistic regression analysis was conducted to explore the influencing factors of cerebral microbleeds. Receiver operating characteristic (ROC) curves were plotted to assess the predictive value of serum OPG and calcium for cerebral microbleeds. Results Significant differences were observed in age, proportions of patients with drinking and hypertension as well as diabetes, systolic blood pressure, diastolic blood pressure, and serum OPG and calcium levels between the cerebral microbleed group and the non-cerebral microbleed group (P < 0.05). Multivariate Logistic regression analysis revealed that older age, history of alcohol consumption, history of hypertension, high systolic blood pressure, and high OPG level were independent risk factors for cerebral microbleeds (OR=1.480, 1.330, 1.843, 1.632, 1.652; P < 0.05), while high calcium level was an independent protective factor for cerebral microbleeds (OR=0.721, P < 0.05). The AUC values for the prediction of cerebral microbleeds in acute ischemic stroke patients using serum OPG or calcium alone, and their combination were 0.853, 0.825, and 0.921, respectively, with the combined prediction showing higher value than the individual prediction (Z=2.895, 3.138; P < 0.05). Spearman rank correlation analysis revealed a positive correlation between serum OPG and the severity of cerebral microbleeds (r_s=0.736, P < 0.05) and a negative correlation between calcium and the severity of cerebral microbleeds (r_s=-0.752, P < 0.05). No significant differences were observed in serum OPG and calcium levels between patients with cerebral microbleeds in different locations (P > 0.05). Conclusion Elevated serum OPG and reduced calcium levels are observed in patients with acute ischemic stroke and cerebral microbleeds. The levels of serum OPG and calcium are closely related to the severity of cerebral microbleeds, and early combined detection can effectively predict the risk of cerebral microbleeds.

Cerebral infarction, with high incidence, high mortality, high disability and high recurrence rates, can impose a serious burden on families and society. After cerebral infarction occurrence, neurons, as the fundamental structures of the central nervous system, are unable to renew or multiply after death; hence, full recovery from neurological impairments following cerebral infarction is challenging. With stem cell and genetic recombination advancements, cellular replacement therapy after cerebral infarction progresses, which helps clinical transformation and application. In this paper, the basic researches of cellular replacement therapy after cerebral infarction are reviewed from 3 aspects: endogenous nerve regeneration, exogenous stem cell transplantation, and in situ somatic cell trans-differentiation into neurons, in order to provide references for cerebral infarction treatment

Objective:To investigate clinicopathological and magnetic resonance imaging (MRI) characteristics of pediatric cardiac tumors.Methods:The clinical, pathological and MRI data of 7 patients with pediatric cardiac tumors confirmed by pathological examination in Children's Hospital of Chongqing Medical University from February 2012 to December 2016 were retrospectively analyzed.Results:There were 3 males and 4 females with first diagnosis age ranging from 1 month to 3 years. As for clinical presentation, most cases were featured with cardiac murmur and enlarged cardiac boundary; only 1 case had acute cerebral infarction, and 1 case did not show any abnormal performance. Pathological findings showed that 6 cases of benign tumors (including 2 cases of fibroma, 1 case of rhabdomyoma, 1 case of myxoma, 1 case of lipoma and 1 case of hemangioma), 1 case of malignant tumor (primitive neuroectodermal tumor of pericardium). MRI results showed that the signal intensity of malignant tumor was higher than that of normal myocardium in each sequence; significant differences were found in benign tumors; first-pass perfusion, cardiac cine image and late gadolinium enhancement were the most obvious.Conclusions:The clinical presentations of pediatric cardiac neoplasms are atypical. Each tumor type has pathognomonic pathological features. MRI has great advantages in the diagnosis and differential diagnosis of cardiac tumors especially for benign tumors.

【Objective】 To determine the reference range of thromboelastogram(TEG) and establish a TEG feature for local population by measuring TEG parameters in healthy adults in Shenzhen comparing the difference between gender and age, and analyzing the reference data provided by reagent manufacturer. 【Methods】 A total of 916 healthy adults, aged between 19 to 59, who did their regular health checks in our hospital from September 2020 to August 2021 were selected. The TEG(from Lepu Medical Technology Co., Ltd.) was performed, and the clot reaction time(R), clot formation time(K), coagulation angle(α-Angle), maximum amplitude(MA), coagulation index(CI), fibrinolysis index LY30 and the estimated percent lysis (EPL) were analyzed. 【Results】 The reference ranges of TEG parameters, including R, K, α-Angle, MA, CI, LY30 and EPL, of 916 healthy adults from Shenzhen were 3.25~8.19 min, 0.66~3.18min, 47.70~76.56deg, 50.05~72.91mm, -4.3~3.4, 0~2.2% and 0~3%, respectively. The value of α-Angle, CI, K, LY30, MA and R didn’t all meet the given range provided by the manufacturer; some were exceeding and some inferior to. A total of 227 out of 916 individuals presented abnormal results, relative to the references, in at least one parameter, and 78 were diagnosed of abnormal coagulation based on the given reference range, with a specificity of 75.2%. 【Conclusion】 The reference range of TEG parameters of Shenzhen locals is significantly different from that provided by manufacturers. And it is imperative for local TEG laboratories to establish their own reference ranges according to age and gender groups based on local population characteristics.

OBJECTIVE: To report on a novel KIR3DL3 allele identified in a southern Han Chinese individual. METHODS: Peripheral blood sample was collected from a voluntary blood donor with inconclusive result by KIR3DL3 sequence-based typing (SBT). Total mRNA was extracted and subjected to reverse transcription to obtain KIR3DL3 cDNA, which was then amplified by PCR with a pair of KIR3DL3-specific primers. The product was subjected to cDNA cloning and sequencing. RESULTS: cDNA cloning and sequencing have identified a wide-type KIR3DL3*00802 allele and a novel KIR3DL3*064 allele. The latter differed from KIR3DL3*00601 by a missense variant at codon 374[c.1184 C>T (p.Thr374Ile)] in exon 9. The novel KIR3DL3 allele has been officially assigned by the KIR subcommittee of World Health Organization Nomenclature Committee for factors of HLA system. CONCLUSION cDNA cloning and sequencing may be used to distinguish inconclusive results in KIR3DL3 SBT in order to identify novel KIR alleles.

Chloroquine (CQ) phosphate has been suggested to be clinically effective in the treatment of coronavirus disease 2019 (COVID-19). To develop a physiologically-based pharmacokinetic (PBPK) model for predicting tissue distribution of CQ and apply it to optimize dosage regimens, a PBPK model, with parameterization of drug distribution extrapolated from animal data, was developed to predict human tissue distribution of CQ. The physiological characteristics of time-dependent accumulation was mimicked through an active transport mechanism. Several dosing regimens were proposed based on PBPK simulation combined with known clinical exposure-response relationships. The model was also validated by clinical data from Chinese patients with COVID-19. The novel PBPK model allows in-depth description of the pharmacokinetics of CQ in several key organs (lung, heart, liver, and kidney), and was applied to design dosing strategies in patients with acute COVID-19 (Day 1: 750 mg BID, Days 2-5: 500 mg BID, CQ phosphate), patients with moderate COVID-19 (Day 1: 750 mg and 500 mg, Days 2-3: 500 mg BID, Days 4-5: 250 mg BID, CQ phosphate), and other vulnerable populations (.., renal and hepatic impairment and elderly patients, Days 1-5: 250 mg BID, CQ phosphate). A PBPK model of CQ was successfully developed to optimize dosage regimens for patients with COVID-19.

Objective To investigate the clinical value of abnormal course of fetal superior mesenteric arteries (SMA) detected by ultrasound during the first trimester (11-13+6 weeks) in predicting congenital diaphragmatic hernia (CDH). Methods This study enrolled women who underwent fetal nuchal translucency (NT) screening during the first trimester in Central Hospital of Panyu District from March to December 2017. Low-speed high-definition flow imaging was used to observe the course of fetal SMA, and it was regarded as abnormal when the angle between SMA and abdominal aorta >90°. Once abnormal course of the SMA was suspected, the position of fetal thoracic cavity and abdominal organs would be scanned carefully. Furthermore, ultrasound examinations would be repeated at 16-18, 20-24, 28-32 and 37-40 weeks of gestation. Fetus diagnosed as CDH by ultrasound would be scheduled for MRI or autopsy to confirm the diagnosis. Pregnancy outcomes of all cases were followed up by telephone. Descriptive statistical analysis was used in this study. Results A total of 6 899 gravidas (6 964 fetuses) underwent NT scan during the first trimester were enrolled and the SMA of all fetuses were successfully displayed. Three cases with abnormal course of the SMA were identified. Two of them were diagnosed with left CDH at 17+ and 23+ weeks of gestation, which was confirmed by autopsy after termination of pregnancy, and the other one terminated pregnancy in first trimester due to a large omphalocele. Among the 6 961 fetuses with normal SMA, the pregnancy outcomes of 6 120 were successfully followed up, only one of which was found to have left CDH at 32 gestational weeks by ultrasound examination, and was later confirmed by neonatal MRI after delivery. No other fetal CDH was detected. Conclusions Abnormal course of the SMA identified in early pregnancy may be a simple and effective indicator for CDH that allow early intervention and treatment.