Objective:To analyze the effect of endoscopic submucosal tunnel dissection on patients with large esophageal superficial neoplasms.Methods: Chosen patients with large esophageal superficial neoplasms in our hospital as research object, randomly divided into control group treated by endoscopic mucosal dissection (ESD) and observation group treated by endoscopic submucosal tunnel dissection(ESTD), compared surgery related indicators, complications and treatment outcomes.Results: 1)Observation group patients’ tumor stripping rate (23.17±4.73)/min was significantly higher than control group patients’ (12.65±2.19)/min; Intraoperative blood loss(9.14±0.67)ml, total length of hospital stay (7.34±1.89) d, were significantly less than control group patients with intraoperative blood loss(21.38±3.14)ml, total length of hospital stay (13.21±3.05)d, t value was 4.965, 5.395, 4.932, respectively(t=4.965,t=5.395,t=4.932;P<0.05); 2)Observation group patients esophageal bleeding ESTD rate(5.26%), esophageal stricture rate(31.58%), mediastinal emphysema rate(5.26%), esophageal perforation rate of 0, were significantly less than control group patients with esophageal bleeding rate(31.58%), esophageal stricture rate(5.26%), mediastinal emphysema rate(21.05%), esophageal perforation rate (26.32%), t value were 4.378, 4.378, 4.471, 5.758, (t=4.378,t=4.378,t=4.471,t=5.758;P<0.05); 3)Observation group patients with no recurrence during the follow-up period, control group patients with local recurrence rate of 21.05%, t value 8.623,P<0.05(t=8.623, P<0.05).Conclusion: Endoscopic tunnel mucosal stripping technique can effectively improve the complete tumor removal rate, during the process of optimization operation at the same time reduce the occurrence of complications, to the improvement of the prognosis of patients with positive clinical significance.

OBJECTIVE:To establish a method for determination of trace platinum in zolpidem tartrate. METHODS:Graphite furnace atomic absorption spectrometry(GFAAS)was adopted for direct determination after dissolving sample in 1% Hydrochloric acid solution. Horizontal platform graphite tube was performed with detection wavelength of 244.79 nm,slit width of 0.2 nm and current intensity of hollow cathode lamp of 6 mA. The mode of background correction was zeeman effect,the peak height was used as measurement pattern,and the injection volume was 20 μl. RESULTS:The linear range of platinum was 0-100 ng/ml(r=0.999 0);RSDs of precision and reproducibility tests were no more than 2.0%;recovery was 97.78%-103.07%(RSD=1.6%,n=9);the detection limit was 1.48 ng/ml. CONCLUSIONS:This method is simple and rapid with good precision and accuracy,and can be applied for the determination of trace platinum in zolpidem tartrate.

Objective:To investigate methods of molecular diagnosis and clinical features of 46, XY disorders of sexual development(DSD).Methods:A total of 206 cases of 46, XY DSD patients, who visited the Shanghai Ninth People′s Hospital, Shanghai Jiaotong University School of Medicine, from July 2009 to June 2021, underwent AA chip based on multiplex PCR and probe-capture-targeted next-generation sequencing. Clinical features of patients with genetic diagnosis were analyzed.Results:Among 206 patients, the diagnostic rate of patients with micropenis, hypospadias and cryptorchidism was the highest, up to 75.28%. Almost all patients had different degrees of undermasculinized external genitalia. The most frequent phenotype was micropenis with hypospadias(87.25%). Only one gene variant was detected in 81 patients(39.32%), multiple genetic variants were detected in 104 patients(50.49%), and no gene variant was identified in 21 patients(10.19%). 107 patients had definite genetic diagnosis, with a diagnostic rate of 51.94% by adding the pathogenic and likely pathogenic ratios following the American College of Medical Genetics and Genomics(ACMG) guidelines, including 40 patients of steroid 5α-reductase type 2(SRD5A2) variants(37.38%), 36 patients of androgen receptor(AR) variants(33.64%), 13 patients of steroidogenic factor 1(NR5A1) variants(16.82%), 6 patients of 17β-hydroxysteroid dehydrogenases 3(HSD17B3) variants(5.61%), 2 patients of 17α-hydroxylase/17, 20-lyase enzyme(CYP17A1), Wilms′ tumor 1(WT1) and GATA binding protein 4(GATA4) variants(1.87%), and one patient of luteinizing hormone receptor(LHCGR) variant(0.93%). Gynecomastia was found in 29 of 81 postpubertal patients, of which 25(86.21%) had AR variants.Conclusions:46, XY DSD presents complex clinical manifestations and molecular etiologies. Targeted nextgeneration sequencing has the advantages of high throughput, high efficiency and low cost, which has a high value especially in etiological diagnosis of 46, XY DSD with large genetic heterogeneity.

Objective To establish a method for acquisition, perfusion, preservation and transportation of the genetically modified pig kidneys. Methods An eight genetically modified pig was utilized as experimental subject. Prior to kidneys procurement, the health status of the pig was assessed through hematology examination, and the vascular structure of the kidneys was examined using imaging techniques. Following kidneys acquisition, the pig kidneys were perfused and subsequently packaged into the cryogenic storage container labeled "For Organ Transportation Only" for interprovincial transport after communicating the transportation process with transportation department. To evaluate pathological damage to the pig kidneys, a serious of methods were employed such as hematoxylin-eosin (HE) staining, real-time fluorescent quantitative polymerase chain reaction (RT-qPCR), terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) fluorescence staining and enzyme-linked immune absorbent assay (ELISA). Results The preoperative examination of the eight genetically modified pig showed that the serum creatinine was 73.2 μmol/L, blood urea nitrogen was 2.8 mmol/L and hemoglobin was 116 g/L, all within the normal range, indicating normal renal function. CT angiography revealed no lesions in the pig kidneys, and no dilation, stenosis or premature branching of the blood vessels. The total time of obtaining the left and right kidneys from the eight genetically modified pig was (125 ± 10) min, with a blood loss of (20 ± 2) mL. The warm ischemia times were 3 min and 7 min, respectively. The perfusion and trimming times of the left and right kidneys were 36 min and 41 min, respectively. After perfusion, both kidneys were white and moist. The cold preservation and transportation time was 8 h. HE staining showed that some glomeruli were shrunk, and the lumens of the surrounding renal tubules were slightly depressed and swollen with partial inner membrane shedding and microvacuoles formed when the kidneys were preserved for 8 h. The level of cysteinyl aspartate-specific proteinase-3 messenger RNA in the kidneys tissue gradually increased with the extension of cold preservation time after 2 h (P<0.05). TUNEL fluorescence staining showed that only a small number of cells underwent apoptosis after 8 h of cold preservation, which was not significantly different from that at 0 h (P>0.05). ELISA results showed that the contents of lactate dehydrogenase (LDH) and creatinine in the preservation solution remained relatively stable, but the content of kidney injury molecule 1 (KIM-1) gradually increased with the extension of preservation time, suggesting that the pig kidneys had mild injury. Conclusions By establishing methods for acquisition, perfusion, preservation and transportation of the kidneys from genetically modified donor pig, it is possible to effectively and reliably use genetically modified pig kidneys for xenotransplantation.

The pandemic of coronavirus disease 2019 (COVID-19) is changing the world like never before. This crisis is unlikely contained in the absence of effective therapeutics or vaccine. The papain-like protease (PLpro) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) plays essential roles in virus replication and immune evasion, presenting a charming drug target. Given the PLpro proteases of SARS-CoV-2 and SARS-CoV share significant homology, inhibitor developed for SARS-CoV PLpro is a promising starting point of therapeutic development. In this study, we sought to provide structural frameworks for PLpro inhibitor design. We determined the unliganded structure of SARS-CoV-2 PLpro mutant C111S, which shares many structural features of SARS-CoV PLpro. This crystal form has unique packing, high solvent content and reasonable resolution 2.5 Å, hence provides a good possibility for fragment-based screening using crystallographic approach. We characterized the protease activity of PLpro in cleaving synthetic peptide harboring nsp2/nsp3 juncture. We demonstrate that a potent SARS-CoV PLpro inhibitor GRL0617 is highly effective in inhibiting protease activity of SARS-CoV-2 with the IC