AIM: To establish human lymphocyte cell line secreting monoclonal antibody against Rhesus(D) antigen and analyse the nucleotide and deduced amino acid sequences of a human monoclonal anti-D Fab fragment. METHODS: By using PCR method, the cDNA of human anti-(Rhesus D) antibody(lgM ?)Fab fragment was amplified from an Epstein- Barr-virus-transformed cell line. Cloning and subsequent sequence analysis of the Fab fragment was performed. The deduced amino acid sequence was compared and analysed with previously published sequences. RESULTS : A band of approximate 700 and 650 base pairs was amplified using lgM heavy chain primers and ? light chain primers, respectively. Sequence analysis indicated that the deduced amino acid sequences was in agreement with the characterization of the amino acid present in the human lg Fab fragment. CONCLUSION: The cloning and sequencing of a human anti-Rhesus (D) antibody Fab fragment cDNA will make benefits for production of recombinant anti-Rhesus (D) antibody and prevention of Rh haemolytic disease in the newborn.

Objective:This study aims to explore the causal relationship between obstructive sleep apnea (OSA) and type 2 diabetes (T2D) using bidirectional Mendelian randomization (MR).Methods:The genetic data related to OSA were obtained from the FinnGen Biobank (Ncase=16, 761, Ncontrol=201, 194) in the Genome-wide association study (GWAS). Three single nucleotide polymorphism (SNP) were screened out as instrumental variable (IV) of OSA. The genetic data related to T2D were derived from a large Meta-analysis of GWAS (Ncase=62, 892, Ncontrol=596, 424), 114 SNP were selected as IV of T2D. Multiple MR methods were used for analysis and inverse variance weighted (IVW) was performed as main method. The sensitivity of MR analytic results was analyzed using MR-Egger and other methods, and the IV was evaluated using F-value statistics. Results:MR analysis showed that OSA was significantly associated with increased risk of T2D ( OR=2.016, 95% CI: 1.185-3.429, P<0.05). There was no significant relationship between T2D and OSA risk ( OR=1.030, 95% CI: 0.980-1.082, P=0.238). There was heterogeneity in both-way results (OSA?T2D, P=1.808×10 -11; T2D?OSA, P=1.729×10 -7), and no horizontal pleiotropy (OSA?T2D, P=0.477; T2D?OSA, P=0.349). IV of OSA and T2D-selected in the study were strong instrumental variables ( F statistics of OSA=20.543; F statistics of T2D=30.117). Conclusion:Our results supported that OSA was a risk factor for T2D, but T2D had no significant impact on the incidence of OSA. Blood glucose monitoring and diabetes screening in OSA patients might be beneficial to the early detection and intervention of T2D.

Objective To investigate the value of anti-HI antibody screening and identification in clinical blood transfusion . Methods 4 cases of anti-HI antibody positive in our hospital from February 2016 to November were selected as the research subjects .The irregular antibodies screening and identification were performed by adopting the anti-human globulin and saline test tube method ,then the blood transfusion effect was evaluated .Results Four cases were ABO blood group ,in which 2 cases were group A RhD positive and 2 cases were group AB RhD positive .The screening results of anti-human globulin method were weak positive , while which of saline test tube method were positive .Under the saline medium condition ,the reaction results of serum with 16 spectrum cellular reaction in 4 cases were positive ,while which with autoerythrocytes were negative ;under the anti-human globulin medium condition ,the reaction results of serum with 16 spectrum cellular reaction in 4 cases were negative except 1 case of weak positive ,while which with autoerythrocytes were negative .After blood transfusion in 2 cases of treatment blood use ,Hb increase reached the expected effect without blood transfusion adverse reactions occurrence .Conclusion In blood transfusion for the patients with anti-HI antibody positive ,the blood of same type ABO and compatible results in both saline test tube method and anti-human globulin tests should be selected .

Objective To summarize and analyze the pathological data of 431 cases of renal biopsy from Quzhou, Zhe-jiang and to discuss the epidemiological characters via pathological distribution in Quzhou. Methods A total of 431 patients with kidney diseases hospitalized between January 2012 and December 2014 by our hospital were reviewed and their epidemiological characters and distribution of pathological patterns were analyzed. Results Among the 431 speci-mens, 181 cases were from males (42.0%), and 250 were females (58.0%, male/female=1:1.38). As to the clinical types of disease, the most common disease was nephrotic syndrome (NS, 178 cases, 41.3%), followed by chronic glomeru-lonephritis (CGN, 142 cases, 32.9%). As to the etiologic classification, a majority of 342 cases were primary glomeru-lonephritis (PGN, 79.3%), 77 cases were secondary glomerulonephritis (SGN, 17.9%), 11 cases were tubulo-interstitial disease(TID, 2.6%), and 1 case was renal transplantation disease(0.2%). Among the PGN patients, IgA nephropathy (IgAN) and membranous nephropathy(MN) were most commonly observed, accounting for 32.7%and 24.5%, respectively. Of the SGN patients, lupus nephritis(LN) was most common, accounting for 40.2%, which was followed by Henoch-Schonlein purpura nephritis (HSPGN, 18.2%) and hepatitis b virus-associated glomerulonephritis (HBVGN, 13.0%). Conclusion PGN is the most common etiological factor of kidney biopsy, with IgAN and MN at most. LN is the main type of SGN, followed by HSPGN and HBVGN. In Quzhou, Zhejiang, the incidence rate is higher in young adults, indi-cating that special attention should be paid on IgAN, MN and LN in prevention and treatment of kidney diseases.

OBJECTIVE：To stu dy the improvement effect s of Dantaojin （Salvia miltiorrhiza ，Persicae Semen ，Curcumae Radix）extract on oxidative stress of liver and kidney in chronic lead poisoning model mice. METHODS ：Totally 72 mice were randomly divided into normal control group ，model group ，positive control group （dimercaptosuccinate，70 mg/kg），Dantaojin extract low-dose ，medium-dose and high-dose groups （20，40，60 g/kg），with 12 mice in each group. Except for normal control group，other groups were given intraperitoneal injection of lead acetate solution 20 mg/kg（every other day ，consecutive 20 days） to establish chronic lead poisoning model. After modeling ，administration groups were given relevant medicine intragastrically ， normal control group and model group were given constant volume of water intragastrically ，once a day ，for consecutive 20 days. After last medication ，body weight of mice was weighed ，and organ coefficients （kidney and liver ）were calculated. Serum levels of ALT ，AST，BUN and Scr were determined by automatic biochemical detector. HE staining was used to observe histopathological features of liver and kidney. ELISA method was used to determine the levels of GSH-Px ，SDO and MDA in liver and renal tissue. RESULTS：Compared with normal control group ，body weight （except for high-dose group ），the levels of SOD （except for high-dose group ）and GSH-Px were all decreased significantly in model group and Dantaojin extract groups ，while the renal coefficients（except for high-dose group ），liver coefficients （except for low-dose ，medium-dose and high-dose group ），the levels of BUN （except for high-dose group ），Scr，AST（except for high-dose group ），ALT and MDA were increased significantly （P＜ 0.05 or P＜0.01）. The epithelial cells of glomerulus and renal tubules were atrophied ，the arrang ement of hepatocytes was loose and some cells were necrotic. Compared with model group ，body weight ，the levels of SOD and GSH-Px were increased significantly in positive control group and Dantaojin extract groups，while the renal and liver coefficients ，the levels of BUN，Scr，AST，ALT and M DA were decreased significantly（P＜0.05 or P＜0.01）. Histopathological fea tures of liver and renal tissue were improved significantly in Dantaojin extract medium-dose and high-dose groups. CONCLUSIONS ：Dantaojin extract could improve oxidant stress injury in liver and renal tissue，the mechanism of which may be associated with eliminating reactive oxygen radicals ，inhibiting lipid peroxidation and enhancing antioxidant defense ability.

Objective:To analyze the clinicopathological features of histological mixed-type early gastric cancer and to compare the endoscopic pathological features of dominant proportions.Methods:Clinical data of 43 patients with histological mixed-type early gastric cancer who underwent endoscopic submucosal dissection (ESD) at the Department of Gastroenterology, Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School from January 2015 to December 2020 were retrospectively analyzed. The endoscopic characteristics, coincidence rate of preoperative diagnosis and postoperative pathology of the patients were analyzed. The endoscopic pathological findings of differentiation dominant group and undifferentiation dominant group were compared.Results:Histological mixed-type early gastric cancer was more common in patients over 60 years old (55.81%, 24/43) , male (65.12%, 28/43), and Helicobacter pylori ( HP) infection (69.77%, 30/43). Most lesions were found in atrophic background (79.07%, 34/43), and lower part of stomach (65.12%, 28/43). Redness or mixed redness was the main color (76.74%, 33/43), and 0-Ⅱc was the main type in lesion morphology (58.14%, 25/43). Characteristics of both differentiated and undifferentiated carcinoma could be observed under magnifying endoscopy. Only 16.28% (7/43) of the patients were diagnosed as having histological mixed-type carcinoma by biopsy. Postoperative pathology showed that there were 33 cases in differentiation dominant group and 10 cases in undifferentiation dominant group. There were 15 (34.88%) patients with deep submucosal infiltration, and 3 (6.98%) patients with positive lymphatic vessels. Differentiation dominant group had clearer demarcation line under endoscopy [78.79% (26/33) VS 4/10, χ2=7.753, P=0.012]. There was no significant difference between the two groups in gender, HP infection, background mucosa, lesion location, endoscopic morphology, lesion long diameter, color, or microscopic and microvascular manifestations under magnifying endoscopy ( P>0.05). Tub2+por was the most common pathological type in differentiation dominant group (78.79%, 26/33), and sig+tub2 was the dominant combination in undifferentiation dominant group (6/10). Endoscopic diagnosis of differentiated primary carcinoma was highly sensitive (93.94%, 31/33), and highly specific for undifferentiated primary carcinoma (94.28%, 33/35). Conclusion:Histological mixed-type early gastric cancer can be diagnosed qualitatively by endoscopy. It is difficult to accurately judge the differentiation advantage and the invasion depth before the operation, which is prone to postoperative pathological upgrading. Therefore, the indications should be strictly considered for ESD treatment for mixed-type early gastric cancer.

Objective:To explore the clinical value of prenatal ultrasound in diagnosis of fetal cleft lip and palate during first-trimester (11-13 + 6 gestational weeks). Methods:Ultrasonographic images were retrospectively selected from those fetuses who underwent first trimester scanning during July 2017 to June 2020 in the Affiliated Suzhou Hospital of Nanjing Medical University. Fetal facial mid-sagittal section and the retronasal triangle (RNT) section were combined together to evaluate whether the fetuses had cleft lip and palate (CLP) or not. All fetuses were followed up to birth or induced abortion.Results:A total of 5 520 fetuses were enrolled, with crown-rump length (CRL) between 45-84 mm. Seven cases of different types of CLP were detected by the 2 combined sections, including 4 cases with unilateral CLP, 1 case with median CLP, and 2 cases with bilateral CLP, which were confirmed by follow-up. In addition, 2 cases of isolated cleft lip (CL) were missed.Conclusions:Combination of fetal facial mid-sagittal section and RNT section is useful for the early diagnosis of fetal cleft lip and palate during first-trimester scanning.

Objective:To explore the relationship between soft markers found in the first trimester (11-13 + 6 gestational weeks) ultrasound screening and fetal adverse pregnancy outcomes. Methods:Single pregnancy fetuses were selected from the Multicenter Clinical Study of First Trimester Screening in China during August 2017 to August 2020. The types and detection rate of soft markers during the first trimester were compared. The correlation between positive soft markers and adverse pregnancy outcomes was analyzed by binary Logistics regression.Results:A total of 16 625 fetuses with complete follow-up outcomes were included in the group. Six hundred and seven ultrasonic soft markers were detected in 556 fetuses with positive soft markers during the first trimester, and the first four most frequently occurred were increased nuchal translucency (NT) (2.08%, 345/16 625), echogenic intracardiac focus (EIF) (0.94%, 156/16 625), hypoplasia of fetal nasal bone (0.20%, 34/16 625), single umbilical artery (SUA) (0.19%, 31/16 625). Among 556 fetuses, the incidence of adverse pregnancy outcome in fetuses with two or more positive soft markers was 32.50% (13/40), which was significantly higher than fetuses with single positive soft marker (11.05%, 57/516), and the difference was statistically significant (χ 2=5.055, P<0.001). The incidence of adverse pregnancy outcome in positive soft markers fetus associated with structural abnormalities was 80.77% (21/26), which was significantly higher than fetuses with isolated positive soft marker (12.08%, 64/530), and the difference was statistically significant (χ 2=90.310, P<0.001). Binary logistic regression analysis showed choroid plexus cyst (CPC), SUA, echogenic bowel (EB), absent/reversed a-wave of ductus venosus, hypoplasia of fetal nasal bone, increased NT, and EIF were closely related to the adverse pregnancy outcomes (all P<0.05). However, there were no significant correlations between tricuspid regurgitation (TR), pyelectasis (PYE) and fetal adverse pregnancy outcomes (all P>0.05). Conclusions:The ultrasonic soft markers during the first trimester are of great significance in predicting fetal adverse pregnancy outcomes. For multiple positive soft markers or positive soft markers combined with structural abnormalities, more attention should be paid to them and comprehensive evaluation is required to be carried out.

Objective:To investigate the metabolism-related proteins and their presence in the plasma of female androgenetic alopecia (FAGA) patients.Methods:From March 2021 to March 2023, FAGA patients aged 18-50 (FAGA group) and healthy women (HC group) were recruited from the Dermatology Outpatient Department of Huashan Hospital. 3 ml of peripheral venous blood was collected from each participant and centrifuged to obtain plasma. Olink proteomics analysis was performed on the collected plasma, differentially expressed proteins were screened with R language, the diagnostic accuracy of the differentially expressed proteins was assessed using receiver operating characteristic (ROC) curve. Gene ontology (GO) analysis was performed on differentially expressed proteins. Immunofluorescence analysis on hair follicles in the parietal region of the FAGA group and the occipital region of the HC group was performed to validate the differentially expressed proteins identified. SPSS 25.0 software was used to analyze the data, with normal distribution metric data represented by Mean±SD. Student’s t-test was used to compare the basic information of two groups of subjects and the relative fluorescence intensity of differentially expressed proteins in hair follicles. Pearson correlation analysis was performed on plasma metabolism-related proteins and the basic information of subjects. P<0.05 indicates a statistically significant difference. Results:Sixty-one cases were included in the FAGA group, with an average age of (33.8±7.4) years and an onset age of (29.5±7.8) years. Among them, 38 cases were mild FAGA, 14 cases were moderate, and 9 cases were severe. Twenty-seven cases were included in the HC group, with an average age of (32.0±7.7) years. There was no statistically significant difference in the basic information (age, body mass index, testosterone, 25-hydroxyvitamin D, uric acid, and ferritin levels) between the two groups of subjects ( P>0.05). Compared to the HC group, the plasma of the FAGA group showed 26 significantly upregulated differentially expressed proteins ( P<0.05), with AHCY and NECTIN2 exhibiting the most significant differences (all P=0.003). The ROC curve evaluation revealed that the area under the curve for AHCY and NECTIN2 was greater than 0.7, indicating good diagnostic accuracy. The GO analysis revealed that the differentially expressed proteins were primarily enriched in the BAT3 complex (cellular component), ubiquitin-dependent ERAD pathway, natural killer cell activation (biological process), as well as ubiquitin protein ligase binding and ubiquitin-specific protease binding (molecular function). Pearson correlation analysis revealed that AHCY ( r=-0.23, P=0.010) and NECTIN2 ( r=-0.31, P=0.033) were negatively correlated with the severity of hair loss in FAGA patients. The results of hair follicle immunofluorescence analysis showed that the relative fluorescence intensity of AHCY and NECTIN2 in the FAGA group was higher than that in the HC group ( P<0.05). In other words, both AHCY and NECTIN2 were upregulated in the FAGA group. Conclusion:Metabolism-related proteins play an important role in FAGA. AHCY and NECTIN2 may serve as early diagnostic biomarkers for FAGA.