Objective To analyze the peculiarity of infants steroid-resistant nephrotic syndrome (SRNS) and to assess the efficacy,side-effect and relapse of various of Tacrolimus prescribed in infants SRNS.Methods A total of 76 case of infant SRNS from August 2012 to August 2015 in Guangzhou Weman and Children's Medical Center grouped into oral Tacrolimus (TAC group),Methyprenisolone pulse therapy (MP group) and Methyprenisolong combined Cyclophosphamide(CTX) pulse therapy(MP + CTX group),were observed for 1 year,and the urine protein excretion,renal function (CCr),blood glucose (B G),urine retinal-binding-protein (URBP),lymphocyte count etc.were recorded and the situation of infection and relapse regularly were monitored regularly.The data were retrospectively analyzed by the statistical method.Results All SRNS children underwent kidney biopsy,and 36 cases of minimal change disease,32 cases of mesangial proliferative glomerulonephritis and 8 cases of focal segmental glome-rulosclerosis were contained in the patients.The pathological constituent ratios were not obviously different among these 3 groups.By 6-month follow-up,the complete remission ratio of TAC group was 63.64％,the total remission ratio was 95.45％,which were remarkably higher than those of MP group (26.09％,60.87％) and MP + CTX group (41.94％,74.19％);the urine protein excretion of TAC group [(7.8 ± 8.6) mg/(kg · d)] was distinctly lower than that of pretreatment and lower than that of MP group [(144.2 ± 118.3) mg/(kg · d)],and lower than that of MP + CTX group [(91.3 ± 87.4) mg/(kg · d)],and the difference was significant (F =22.69,P ＜ 0.05).The remission time of TAC group was about 2 months,that of other two groups was about 3 months.By 1-year follow-up,the lymphocyte counts including total T-cell (CD3 +),the helper T-cell (CD4 +) and the inhibited T-cell (CD8 +) of TAC group decreased obviously(all P ＜ 0.01),which were extremely lower than those of the M P group and MP + CTX group,and there were significant differences (all P ＜ 0.05).By 1-year follow-up,the person-time of infection existed superior to the other 2 groups,TAC group was compared with MP plus group,the rank sum was 348.5 (U =-3.69,P ＜ 0.01);compared with MP + CTX plus group,the rank sum was 369.5 (U =-4.18,P ＜ 0.01).During the observation the URBP of TAC group was distinctly higher than that of the MP group and the MP + CTX group [(13.77 ± 19.19) mg/L vs.(2.50 ± 1.77) mg/L,(2.06 ±3.63) mg/L],and the differences were significant(t =3.16,2.99,all P ＜0.05);the TAC group with BG and CCr maintained stably.Conclusions Tacrolimus shows its own advantages of more reliable effect and less side-effect in the infants with SRNS over MP therapy and MP combined CTX therapy,but it could not lessen the recurrence of the disease,and its long-term prognosis is still not very clear.

Objective To detect specific cell killing effect of radiation combined with horseradish peroxidase (HRP)/indole-3-acetic (IAA) suicide gene therapy controlled by a novel radio-inducible and cancer-specific chimeric gene promoter in lung cancer. Methods We constructed a plasmid expressing HRP enzyme under the control of chimeric human telomerase reverse transcriptase (hTERT) promoter carrying 6 CArG elements, a plasmid expressing HRP enzyme under the control of hTERT promoter carrying single CArG element, and two control plasmids, which named pE6-hTERT-HRP, phTERT-HRP, pControl-HRP, and pControl-luc, respectively. After radiation, the proliferation inhibition and apoptosis induction effect of each type of plasmid in lung cancer cells (A549, SPC-A1) and normal lung cells (hEL) was detected by cell counting and Annexin V-FITC staining. The change of radiosensitivity of lung cancer cells with plasmid system was also detected by clonogenic assays. Results After a single dose radiation of 6 Gy,the average proliferation inhibition rates of pE6-hTERT-HRP, phTERT-HRP, pControl-HRP, and pControl-luc systems were 72. 92% ,40.60% , 51.00% and 25.19% (F= 67.31 , P< 0.01) in A549 cells ,64.63%,30.02%,48.23% and 23.16% (F=64.94, P< 0.01) in SPC-A1 cells, and 20.81%,18.05%, 44.20% and 18.32% (F=52. 19,P<0.01) in normal hEL cells, respectively. The average early apoptosis rates of these four plasmid systems were 36. 63%, 22. 30%, 24. 33% and 12. 53% (F =50. 99,P <0. 01) in A549 cells, 33.73%, 17. 37%, 22. 43% and 11.20% (F = 20. 76, P < 0. 01) in SPC-A1 cells, and 13.53 %, 12. 5%, 21.93% and 12. 16% (F = 15. 08, P < 0. 01) in normal hEL cells,respectively. The sensitizing enhancement ratios of the four plasmid systems were 3.45, 2. 29, 3.05 and 1.21 in A549 cells, while 2. 68, 2. 15, 3.05 and 1.21 in SPC-A1 cells, respectively. Conclusions The new suicide gene system controlled by chimeric promoter may provide a novel therapeutic modality for lung cancer.

OBJECTIVE: To construct expression vector for the SEA-EGF fusion gene. METHOD: Clone the SEA gene and the EGF gene segment with PCR and RT-PCR independently, and connect this two genes by the bridge PCR. Insert the fusion gene EGF-SEA into the expression vector PET-44. Induced the secretion of the fusion protein SEA-EGF by the antileptic. RESULT: The gene fragment encoding EGF and SEA mature peptide was successfully cloned. The fusion gene EGF-SEA was successfully constructed and was inserted into expression vector. CONCLUSION The new recombinant expression vector for fusion gene EGF-SEA is specific for head and neck cancer, laid the foundation for the further study of fusion protein SEA-EGF targeting immune therapy in head and neck tumors.
Enterotoxins ; genetics ; Epidermal Growth Factor ; genetics ; Genetic Vectors ; Head and Neck Neoplasms ; drug therapy ; Humans ; Molecular Targeted Therapy ; Recombinant Fusion Proteins ; biosynthesis ; genetics

A large-scale Class A tertiary comprehensive hospital in Hunan Province,as one of the first pilot units for pre-hospitalization in the province,has pioneered the exploration of a pre-hospitalization management model based on intensive bed management.Through measures such as streamlining the pre-hospitalization process,implementing refined services,establishing dedicated examination channels,and utilizing information technology for comprehensive management of pre-hospitalization pa-tients,the hospital has ensured the smooth implementation of pre-hospitalization work.Practical results have shown a significant reduction in the average length of hospital stay in surgical wards.For instance,the average length of stay in the Department of Breast Surgery and the Department of Otolaryngology decreased by 4.41 days and 3.29 days,respectively,when compared before and after implementation.This study will delve into the impact of the pre-hospitalization management model on reducing the aver-age length of hospital stay,providing a reference for large-scale hospitals in China to improve operational efficiency and alleviate the shortage of hospital beds.It also aims to offer insights into enhancing patient experience before hospitalization and improving overall patient satisfaction as part of initiatives to improve healthcare services.

Objective:To prepare 68Ga-2-(4, 7-bis(carboxymethyl)-1, 4, 7-triazonan-1-yl)pentanedioic acid (NODAGA)-YHWYGYTPQNVI (GE11) and evaluate its feasibility of PET imaging for pancreatic cancer. Methods:GE11 peptide was conjugated with NODAGA and then labeled with 68Ga. The labeling yield, radiochemical purity, hydrophilicity, stability and specificity in vitro were determined. Human pancreatic cancer BxPC3 nude mice models ( n=9) were established. MicroPET imaging was then obtained after 30 and 90 min, and mice were sacrificed at 90 min to acquire the radioactivity distribution of main organs and tumors. Pair t test was used to analyze the data. Results:The labeling yield was (73.5±5.4)% and radiochemical purity was more than 98%. After incubation 120 min in mouse serum at 37 ℃, radiochemical purity was more than 92%. The uptake was specific in BxPC3 cell lines. MicroPET images showed that 68Ga-NODAGA-GE11 could accumulate quickly in tumor. Value of tumor uptake was significantly higher than that of normal pancreas at 90 min ((1.38±0.25) vs (0.49±0.07) %ID/g; t=12.67, P<0.05), and the radio-uptake of blood, muscle and bone was lower than that of tumor. Conclusions:68Ga-NODAGA-GE11 is easy to be prepared with high radiochemical purity and good stability, and can specifically target BxPC3 xenograft tumor. However, due to the high uptake in the kidneys and liver, the value of 68Ga-NODAGA-GE11 in PET imaging for pancreatic tumor needs further study.

Animals ; Asian Continental Ancestry Group ; Chickens ; China ; epidemiology ; Cytokines ; metabolism ; Genetic Variation ; Genotype ; Humans ; Influenza A Virus, H7N9 Subtype ; classification ; genetics ; pathogenicity ; Influenza A Virus, H9N2 Subtype ; genetics ; Influenza in Birds ; transmission ; virology ; Influenza, Human ; ethnology ; transmission ; virology ; Mice, Inbred BALB C ; Molecular Sequence Data ; Orthomyxoviridae Infections ; metabolism ; mortality ; virology ; Phylogeny ; Seasons ; Survival Rate ; Virulence ; genetics

Objective:To explore the feasibility of deep learning technology for renal artery recognition in retroperitoneal laparoscopic renal surgery videos.Methods:From January 2020 to July 2021, the video data of 87 cases of laparoscopic retroperitoneal nephrectomy, including radical nephrectomy, partial nephrectomy, and hemiurorectomy, were retrospectively analyzed. Two urological surgeons screened video clips containing renal arteries. After frame extraction, annotation, review, and proofreading, the labeled targets were divided into training set and test set by the random number table in a ratio of 4∶1. The training set was used to train the neural network model. The test set was used to test the ability of the neural network to identify the renal artery in scenes with different difficulties, which was uniformly transmitted to the YOLOv3 convolutional neural network model for training. According to the opinion of two senior doctors, the test set was divided into high, medium, and low discrimination of renal artery and surrounding tissue. High identification means a clean renal artery and a large exposed area. For middle recognition degree, the renal artery had a certain degree of blood immersion, and the exposed area was medium. Low identification means that the exposed area of the renal artery was small, often located at the edge of the lens, and the blood immersion was severe, which may lead to lens blurring. In the surgical video, the annotator annotated the renal artery truth box frame by frame. After normalization and preprocessing, all images were input into the neural network model for training. The neural network output the renal artery prediction box, and if the overlap ratio (IOU) with the true value box was higher than the set threshold, it was judged that the prediction was correct. The neural network test results of the test set were recorded, and the sensitivity and accuracy were calculated according to IOU.Results:In the training set, 1 149 targets of 13 videos had high recognition degree, 1 891 targets of 17 videos had medium recognition degree, and 349 targets of 18 videos had low recognition degree. In the test set, 267 targets in 9 videos had high recognition degree, 519 targets in 11 videos had medium recognition degree, and 349 targets in 18 videos had low recognition degree. When the IOU threshold was 0.1, the sensitivity and accuracy were 52.78% and 82.50%, respectively. When the IOU threshold was 0.5, the sensitivity and accuracy were 37.80% and 59.10%, respectively. When the IOU threshold was 0.1, the sensitivity and accuracy of high, medium and low recognition groups were 89.14% and 87.82%, 45.86% and 78.03%, 32.95%, and 76.67%, respectively. The frame rate of the YOLOv3 algorithm in real-time surgery video was ≥15 frames/second. The false detection rate and missed detection rate of neural network for renal artery identification in laparoscopic renal surgery video were 47.22% and 17.49%, respectively (IOU=0.1). The leading causes of false detection were similar tissue and reflective light. The main reasons for missed detection were image blurring, blood dipping, dark light, fascia interference, or instrument occlusion, etc.Conclusions:Deep learning-based renal artery recognition technology is feasible. It may assist the surgeon in quickly identifying and protecting the renal artery during the operation and improving the safety of surgery.

Objective:A multi-center and large sample volume study was conducted on the verification and improvement of the early established criteria for intelligent routine urinalysis validation (including the microscopic review rules and manual validation rules, referred to as intelligent criteria for short), in order to improve the clinical application of this intelligent criteria.Methods:A total of 31 456 urine specimens were collected from the inpatients and outpatients in six hospitals in China, from March to September 2019. Firstly, 3105 specimens were analyzed for preliminary verification and improvement of the intelligent criteria based on the results of the microscopic examination and manual validation. Secondly, 28 351 specimens were used to verify the clinical application of the improved intelligent criteria. All samples were manually validated as reference.Results:The approval inconsistency rate of the manual validation rules in the original intelligent criteria was 8.59% (202/2 352), and the interception inconsistency rate was 8.84% (208/2 352). The false negative rate and the microscopic review rate of the microscopic review rules were similar to the previous results. Based on an in-depth analysis of big data and the discussions by senior technicians from eight hospitals, one microscopic review rules and four manual validation rules were added, meanwhile two manual validation rule was deleted. The manual validation standards were unified. Finally, the intelligent criteria was improved. Based on the improved intelligent criteria, for microscopic review rules, the false positive rate, false negative rate (misdiagnosis rate), and microscopic review rate did not change significantly, which were 14.72% (457/3 105), 4.06% (126/3 105), and 24.73% (768/3 105), respectively. The approval inconsistency rate and the interception inconsistency rate of manual validation rules were both reduced to 0; the total manual validation rate of the intelligent criteria was 50.89% (1 580/3 105), and the auto-validation rate was 49.11% (1 525/3 105). The large sample volume verification results were consistent with the preliminary verification results of the improved intelligent criteria.Conclusion:This multi-center and large sample volume study had shown that the improved intelligent criteria had better clinical performance.

OBJECTIVE: To establish a non-coated enzyme-linked immunosorbent assay (ELISA) based on zika virus envelope (E) protein for detecting the expression of E protein in infected cells. METHODS: Adherent Vero-143 cells infected with zika virus in a 96-well plate were fixed, and the antibodies against zika virus E protein were added at an optimized concentration to establish the non-coated ELISA method for E protein. The antiviral activities of lignans compound C1 was evaluated using this method. The accuracy of this non-coated ELISA was verified by RT-PCR, and the cross reaction with dengue virus was assessed. RESULTS: After optimization, the background absorbance at 450 nm of uninfected cells was reduced to about 0.20. The antiviral activities of lignans compound C1 detected by this method were basically consistent with the results of RT-PCR. No cross reaction with dengue virus was found in this assay. CONCLUSIONS A non- coated ELISA method based on zika virus E protein was established, which can be used for screening antiviral agents against zika virus.
Antibodies, Viral ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin G ; Immunoglobulin M ; Viral Envelope Proteins ; Zika Virus ; Zika Virus Infection

【Objective】 To explore the feasibility of en-bloc resection of bladder tumors by flexible cystoscope combined with laparoscopic instruments through urethra and to provide reference for the clinical application of this technique. 【Methods】 Self-designed and processed transurethral single-hole PORT and Olympus electronic cystoscope were used as observation mirror; Φ1.8 mm soft grasper, tissue scissors, electric hook, and ultrasonic scalpel were used as instruments; the porcine bladder was used as a model.The PORT was placed through the urethra, and the cystoscope was inserted to observe the inner wall of the bladder and the condition of the mucosa.After the lesion site was identified in the bladder cavity, the soft grasper was inserted to pull the mucosa to be removed, which was then fixed with tension at the target position to maintain a satisfactory feild of view.The surgeon held the cystoscope in the left hand, and operated the laparoscopic instruments into the bladder cavity through the PORT with the right hand.Observing with the cystoscope and lifting and pulling the mucosa with the grasper, the surgeon simulated the cutting and pushing actions to realize the en-bloc resection of the lesioned mucosa. 【Results】 The mucosa at 4 different locations were successfully resected on 2 in vitro porcine bladder models. 【Conclusion】 The in vitro experiments show that the combination of flexible electronic cystoscope and laparoscopic instruments achieves synergistic effects in en-bloc resection of bladder tumor by transurethral single-hole laparoscope without additional iatrogenic bladder injury caused by percutaneous bladder incision.This method is feasible in the treatment of bladder tumors, and has the potential of clinical application after further optimization.