Objective To correctly identify the blood group of ABO and study its molecular biological characteristics.Methods Blood samples from blood donors with discrepancies in forward and reverse typing using the microplate method were subjected to both saline tube agglutination test for serological identification and polymerase chain reaction-sequence specific primers(PCR-SSP)for genotyping.Additionally,direct sequencing of exons 1 to 7 of the ABO gene was performed on some donor samples to analyze their blood phenotype,genotype and gene sequence.Results For 256 samples showing discrepancy between forward and reverse typing in microwell method,119 were identified as normal ABO blood types,90 were weakened ABO antibodies and 47 were ABO subtypes by serology tube test.According to the PCR-SSP genotyping test,233 of 256(91.02%)were consistent with serological phenotype and genotype,17 of 256(6.64%)were inconsistent,and 6 samples can′t read genotype based on the kit result typing table.A total of 17 genotypes were identified in 250 samples as AO1 in 56,AO2 in 58,AA in 50,BO1 in 31,BO2 in 17,BB in 8,O1O1 in 2,O1O2 in 7,AB in 13,AO4 in 1,A205O2 in 1,A205A in 1,A201A in 1,O1O4 in 1,O2O2 in 1,A201B in 1 and A205B in 1.Sequencing of exons 1 to 7 of the ABO gene was carried out in 78 samples,and 29 ABO alleles were detected,seven of which were common alleles(?A101,?A102,?A104,?B101,?O01,?O02,?O04),22 of which were rare alleles(?A201,?A205,?Ax01,?Ax03,?Ax13,?Ax19,?Ax22,?Ael10,?B305,?Bel03,?Bel06/?Bx02,?Bw07,?Bw12,?Bw17,?Bw37,?O05,?O26,?O61,?B(A)04,?B(A)07,?cisAB01 and ?cisAB01var).In addition,six rare allele mutation sites were identified(c.101A＞G;c.103_106delG;c.146_147insGC;c.259G＞T;c.322C＞T;c.932T＞C).Conclusion The identification of ambiguous ABO blood group requires the combination of serological testing and molecular biological examination to correctly identify the blood type and ensure the safety of clinical blood transfusion.

Objective To explore the clinical effect of personalized pars plana vitrectomy(PPV)for proliferative di-abetic retinopathy(PDR).Methods In this retrospective case study,76 patients(86 eyes)diagnosed with PDR and re-ceiving PPV in the Department of Ophthalmology of Songjiang Hospital affiliated to Shanghai Jiao Tong University School of Medicine,from October 2019 to November 2022,were divided into the observation group(40 patients,46 eyes)and the control group(36 patients,40 eyes).Patients in the obseration group were treated with personalized PPV,while patients in the control group were treated with conventional PPV,After treatment,all patients were followed up for 12 months.The operation time,intraoperative use of heavy water and silicone oil,incidence of iatrogenic retinal tears and heavy water resi-dues,proportion of scleral buckling,preoperative and postoperative best corrected visual acuity(BCVA)and intraocular pressure(IOP),retinal reattachment rate at 12 months after surgery,and the incidence of post-vitrectomy vitreous hemor-rhage(PVH),diabetic macular edema(DME)and neovascular glaucoma(NVG)were compared between the two groups.Results The operation time of patients in the observation group was shorter than that in the control group(P＜0.05).Intraoperative use of heavy water and silicone oil in the observation group was lower than that in the control group(both P＜0.05).The incidence of iatrogenic retinal tears and heavy water residues and the proportion of scleral buckling showed no statistically significant difference between the two groups(all P＞0.05).There was no statistically significant difference between the two groups in BCVA preoperatively,3,6 and 12 months postoperatively(all P＞0.05).BCVA in the observa-tion group was better than that in the control group at 1 day,1 week and 1 month after surgery(all P＜0.05).Compared with the preoperative value,BCVA increased in the observation group at 1 day,1 week,1 month,3 months,6 months,and 12 months after surgery(all P＜0.05);in the control group,BCVA increased slightly at 1 day and 1 week(both P＞0.05)and then increased significantly at 1 month,3 months,6 months,and 12 months after surgery(all P＜0.05).The two groups showed no statistically significant difference in IOP at 1 day,1 week,1 month,3 months,6 months,and 12 months postoperatively(all P＞0.05).There was no statistically significant difference in the retinal reattachment rate and the inci-dence of complications such as PVH,DME,and NVG between the two groups at 12 months postoperatively(all P＞0.05).Conclusion Personalized PPV can shorten the operation time,reduce the intraoperative use of heavy water and silicone oil,enhance the efficiency of the operation,and rapidly improve the visual acuity of PDR patients.

Objective:To investigate the clinical characteristics, laboratory characteristics and genetic diagnosis of aromatic L-amino acid decarboxylase deficiency (AADCD), and to improve the understanding of this disease.Methods:Four children diagnosed with AADCD from the Department of Neurology, Beijing Children′s Hospital Affiliated to Capital Medical University from August 2016 to June 2020 were collected, and their clinical manifestations, laboratory and imaging data, and genetic test results were retrospectively analyzed.Results:All the 4 cases were diagnosed in early infancy, with the first symptom of feeding difficulties. They developed paroxysmal dyspraxia accompanied by eye movement crisis, movement regression, hypotonia, growth retardation, sleep disorders and autonomic nervous symptoms such as ptosis, excessive sweating and nasal congestion at the age of 2-4 months, respectively. The 4 children were siblings from 2 families with healthy parents. The dihydroxyphenylalanine decarboxylase ( DDC) gene mutations in cases 1 and 2 were derived from the maternal missense mutation c.1040G>A(P.RG347gln), and from the paternal deletion of exons 11 and 12, respectively. The DDC gene mutation in case 3 was derived from the maternal mutation c.419G>A(p.G140E) and the paternal mutation c.1375C>T(p.H459Y), respectively. Case 4 did not undergo genetic testing. Blood amino acid and acylcarnitine profiles and urine organic acid analyses were performed in 3 cases, and no specific abnormalities were found. In case 3, the results of 3-O-methyldopa (3-OMD) screening by blood dry filter paper increased significantly. Cerebrospinal fluid neurotransmitter detection results showed that the concentrations of 3-methoxy-4-hydroxyphenyldiol, vanillic acid and 5-hydroxyindoleacetic acid were significantly decreased, while the levels of 5-hydroxytryptophan and 3-OMD were increased in case 3. Blood aromatic L-amino acid decarboxylase (AADC) activity decreased significantly in case 3. Cranial magnetic resonance imaging (MRI) and electroencephalogram (EEG) examinations were performed in cases 1, 3, and 4, among which the cranial MRI in case 1 was normal, while the cranial MRI in cases 3 and 4 suggested that myelination was slightly backward. The EEG was normal in all the 3 cases. Cases 1 and 2 died of pneumonia and respiratory failure at the age of 1 year and 10 months. Case 3 was given clonazepam, benxel hydrochloride tablets and vitamin B6 tablets orally after diagnosis at the age of 4 months, and then treated with selegiline hydrochloride tablets and pramexol hydrochloride tablets. At the follow-up of 1 year and 6 months, the frequency of eye movement crisis and movement disorder was reduced, sleep was improved and autonomic nervous symptoms were alleviated, but there was no improvement in developmental delay. Case 4 was diagnosed with cerebral palsy and epilepsy, but failed various antiepileptic drugs and rehabilitation training, and died at the age of 10 due to heart failure and kidney failure. Conclusions:The clinical manifestations of AADCD are complicated and the misdiagnosis rate is high. Infants with early-onset hypotonia, developmental retardation, eye movement crisis, and movement disorders should be screened with dry filter paper as soon as possible for 3-OMD level, and suspicious cases should be diagnosed by cerebrospinal fluid neurotransmitter detection, plasma AADC activity determination, and gene examination. Early diagnosis of AADCD in children and gene mutation carriers can guide treatment and provide genetic counseling to reduce the incidence of the offspring.

By comparing the training status of clinical research methods between United States and China, this article introduces comprehensive training system in the United States in and after the university, and discusses the problems in clinical research methodology training in China. These issues include that when medical students at school do not receive the training of professional clinical research courses, and after they go to the work, they also lack professional and accurate clinical research methodology training, which make it difficult for doctors to independently conduct clinical research. Therefore, it is recommended that Chinese medical schools should systematically establish clinical research methodology courses for undergraduates and graduate students to cultivate the clinical research capabilities of medical students. Secondly, according to the clinical research levels of doctors, different training models are proposed in the study. The existing social resources should be scientifically guided. We hope our work would provide some references for the improvement of clinical research methodology in China, to promote clinicians to be independently responsible for conducting clinical research, and improve the national medical level.

Objective:To investigate the prognostic value of preoperative inflammatory indicators for hepatocellular carcinoma (HCC).Methods:The retrospective cohort study was conducted. The clinicopathological data of 73 patients with primary HCC who underwent radical partial hepatectomy in the Beijing Tsinghua Changgung Hospital of Tsinghua University from December 2014 to July 2019 were collected. There were 57 males and 16 females, aged from 33 to 81 years, with a median age of 58 years. Results of blood examination indicators at the first time in hospital were determined for patients. Observation indicators: (1) the best cut-off values of?? preoperative inflammatory indicators calculated by the maximally selected rank statistics; (2) follow-up; (3) influencing factors for prognosis of HCC patients; (4) comparison of clinicopathological parameters of HCC patients; (5) comparison of predictive value for overall survival. Follow-up was conducted using outpatient examination and telephone interview to determine postoperative survival of patients up to September 2019. Measurement data with normal distribution were represented as Mean±SD, and comparison between groups was analyzed using the t test. Measurement data with skewed distribution were represented as M (range). The best cut-off values ??for continuous variables were obtained using the maximally selected rank statistics based on survival at endpoint of follow-up. Count data were represented as absolute numbers, and comparison between groups was analyzed using the chi-square test or Fisher exact probability. Kaplan-Meier method was used to calculate survival rates, and Log-rank test was used for survival analysis. Univariate analysis was performed using the Log-rank test. Multivariate analysis was performed using the COX proportional hazard model. The time-dependent receiver operating characteristic curve (ROC) was used to compare the predictive value of independent prognostic factors. Results:(1) The best cut-off values of?? preoperative inflammatory indicators calculated by the maximally selected rank statistics: the best cut-off values of neutrophil-lymphocyte ratio (NLR), platelet-lymphocyte ratio (PLR), and prognostic nutrition index (PNI) were 3.46, 131.05, and 45.65. (2) Follow-up: 73 patients were followed up for 31 months (range, 2-57 months). Twenty patients died during the follow-up. (3) Influencing factors for prognosis of HCC patients: results of univariate analysis showed that NLR, PNI, tumor diameter, and tumor differentiation degree were related factors affecting prognosis of patients ( χ2=10.213, 4.434, 5.174, 4.306, P<0.05). Results of multivariate analysis showed that NLR and tumor differentiation degree were independent factors affecting prognosis of patients ( hazzard ratio=4.429, 13.278, 95% confidence interval as 1.662-11.779, 1.056-10.169, P<0.05). (4) Comparison of clinicopathological parameters of HCC patients: of 73 patients, 64 cases had NLR<3.46 and 9 cases had NLR≥3.46. Cases with tumor length >5 cm or ≤5 cm, neutrophils, lymphocytes were 23, 41, (2.9±1.2)×10 9/L, (1.7±0.6)×10 9/L for 64 patients with NLR<3.46, versus 8, 1, (5.8±2.9)×10 9/L, (1.0±0.3)×10 9/L for 9 patients with NLR≥3.46; there were significant differences in above indicators between the two groups ( χ2=7.017, t=2.982, -3.168, P<0.05). (5) Comparison of predictive value for overall survival: time-dependent ROC curves of NLR and tumor differentiation degree for 1-, 2-, 3-, 4-year survival rates had the area under curve of 0.735,0.611, 0.596, 0.574 and 0.554, 0.583, 0.572, 0.556, respectively. NLR had better predictive value for overall survival of patients than tumor differentiation degree. Conclusion:Preoperative NLR is an independent factor affecting prognosis patients, and its predictive efficacy is better than tumor differentiation degree.

【Objective】 To investigate the serological and molecular biological characteristics of para-Bombay phenotype. 【Methods】 ABO blood type, H antigen and Lewis blood type in one blood sample with discrepancy between forward and reverse blood typing were detected by serological method. Antibody screening and identification and cross-match test were also performed by serological method. ABO blood group genes were detected by PCR-SSP, and FUT1 and FUT2 genes were sequenced. 【Results】 The serological test showed that the Para-Bombay phenotype was Ah secretion. The ABO blood group gene was AO2. FUT1 sequencing revealed two mutations: 235G>C and 881_882d^elTT. While FUT2 sequencing showed only one mutation 357C>T. 【Conclusion】 The discovery and accurate identification of blood groups is necessary to ensure the safety of blood transfusion. Blood donors of rare blood groups should be informed and recruited to the team of rare blood donors.

The current epidemic of Corona Virus Disease 2019 (COVID-19) poses a serious threat to people′s health and lives. Following the national policies and deployment, medical community in China invests large amounts of medical resources to help Hubei. However, some hospitals nationwide have suspended outpatient and inpatient services. The authors analyze the underlying causes of suspension of medical services and its harm to patients, and summarize the optimization of disease diagnosis and treatment during the COVID-19 outbreak. In further, they propose reasonable suggestions on safe medical management in general hospitals outside the epidemic area, in order to attract the attention of medical institutions and medical colleagues.

Hilar cholangiocarcinoma (HCCa) is a biliary malignancy that seriously affects the survival of patients.Radical resection is the only cure method for patients to achieve long-term survival.However,due to the low rate of radical resection,its prognosis is poor.In this article,surgical treatment strategies of HCCa in the precision surgery era,3D image evaluation in the curative resection,surgical procedure decision and the preservation of liver parenchyma in the hepatic resection are discussed combined with the research progress at home and abroad.

Pyroptosis is a form of new programmed cell death which is dependent on Caspase-1 in recent years.When it' s stimulated by various dangerous signals from hepatic ischemia-reperfusion injury,the intracellular pattern recognition receptors are assembled into inflammasomes and Caspase-1 which was transformed into active form.Activated Caspase-1 promotes the maturation and secretion of pro-inflammatory cytokines IL-1β and IL-18,initiates the innate immunity rapidly and then induces severe inflammatory reaction.In addition,Caspase-1 can also cleave Gasdermin D and release its N-terminal domain triggering pyroptosis.Many studies showed that pyroptosis play a crucial role in hepatic ischemia-reperfusion injury.In this review,we discussed the activation mechanism and research progress of pyroptosis in hepatic ischemia-reperfusion injury.

The 8th Japan-China Hepato-Pancreato-Biliary Symposium was held in Tokyo,Japan from 22nd to 23rd November 2018.The meeting was convened coincidently with the 80th Annual Congress of Japanese Surgical Association,which attracted the participation of Chinese and Japanese hepatobiliary and pancreatic surgeons.The symposium aimed to explore the latest achievements and clinical issues of diagnosis and treatment for hepato-pancreato-biliary diseases.In this article,authors reviewed the up-to-date research information in order to share the experience,achievements and new information in the field of hepatobiliary and pancreatic diseases with colleagues.