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 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 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 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 observe any effect of repetitive transcranial magnetic stimulation (rTMS) on sleep disorders among children with cerebral palsy (CP).Methods:A total of 102 children with CP and disordered sleep were randomly divided into an experimental group and a control group, each of 51. All were given routine rehabilitation and sleep health education, but the experimental group additionally received rTMS for two weeks. The polysomnography (PSG) results of the two groups were recorded and analyzed.Results:The PSG parameters had improved greatly in both groups after the treatment. The percentage of N2 sleep (depth of sleep during light sleep) in the severe cerebral palsy group and of N3 sleep (depth of sleep during deep sleep) in the moderate cerebral palsy group had increased significantly more than in the mild cerebral palsy group, on average. After the intervention the percentages of N2 and N3 in those with mixed cerebral palsy and of N3 in those with involuntary motor cerebral palsy had increased significantly more than in those with spastic cerebral palsy, on average.Conclusion:rTMS treatment can improve the sleep disorders of children with cerebral palsy, especially N2 sleep among children with moderate to severe cerebral palsy, N3 sleep in cases of mixed or dyskinetic CP.

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 explore the diagnostic value of first-trimester and mid-trimester ultrasound in screening fetal pentalogy of Cantrell, and to analyze missed and misdiagnosed cases.Methods:The fetal ultrasound image characteristics of pentalogy of Cantrell diagnosed in the Affiliated Suzhou Hospital of Nanjing Medical University from March 2018 to November 2022 were retrospectively analyzed. The necessary sections and key features of ultrasound images for diagnosing the disease in first-trimester and mid-trimester were summarized. The diagnostic value of ultrasound screenings in first-trimester and mid-trimester was analyzed, and the progression of the disease during pregnancy was understood, the missed diagnosis rate and misdiagnosis rate were calculated, and the reasons for missing diagnosis were analyzed. All fetuses were followed up to birth or induction of labor.Pentalogy of Cantrell was divided into types Ⅰ, Ⅱ, and Ⅲ according to Toyama′s research.Results:Among the 120 190 fetuses, 13 cases of pentalogy of Cantrell were diagnosed by ultrasound in first-trimester and mid-trimester. Ultrasound predominantly showed the fetal heart being malpositioned outside the chest and the fetal abdominal contents bulging outside the abdominal cavity, and the sternumal echo was partially or completely missed in some cases. All 13 cases were confirmed by follow-up, including 1 case of type Ⅱ, and 12 cases of type Ⅲ. In addition, 1 missed case of type Ⅱ pentalogy of Cantrell was followed up after birth. The correct diagnostic rates of fetal pentalogy of Cantrell using standard ultrasound sections during the first-trimester and mid-trimester were 99.9% and 100%, the sensitivity were 88.9% and 100%, the specificity were both 100%, the positive predictive values were both 100%, and the negative predictive values were 99.9% and 100%, respectively.Conclusions:First-trimester and mid-trimester ultrasound screenings have high diagnostic accuracy for pentalogy of Cantrell, and early detection and early diagnosis are of great clinical significance for the guidance of pregnancy outcomes.