mRNA gene therapy has attracted much attention due to its advantages such as scalability, modification, no need to enter the nucleus and no integration of host genes. In gene therapy, safe and effective delivery of mRNA into cells is critical for the success of gene therapy. In this study, we designed and synthesized an amphiphilic cationic lipopeptide gene vector (dendritic arginine & disulfide bond-containing cationic lipopeptide, RLS) enriched with branched arginine. We achieved a 1.5-fold higher mRNA transfection efficiency in zebrafish compared to the commercial reagent Lipofectamine 2000, and confirmed its good biosafety by in vitro cytotoxicity and in vivo biosafety. First, we characterized the chemical composition of the cationic lipid peptides by nuclear magnetic resonance hydrogen spectroscopy (¹H NMR) and time-of-flight mass spectrometry (MS). The results of particle size and potential tested by dynamic light scattering particle size analysis showed that at a nitrogen/phosphorus (N/P) ratio of 20, the RLS/mRNA composite assemblies formed homogeneous nanoparticles with an average particle size of about 220 nm and a surface ζ potential of about +21 mV. In vitro gene transfection, the transfection experiments demonstrated that RLS exhibited 1.2-fold higher transfection efficiency in human embryonic kidney 293 cells (HEK293) and 3-fold higher transfection efficiency in rat mesenchymal stem cells (MSC) compared to Lipofectamine 2000. In addition, after microinjection of RLS into zebrafish embryos, we evaluated the survival, hatching, and teratogenicity rates, all of which confirmed its favorable in vivo safety profile. Thus, this amphiphilic cationic lipid peptide RLS, enriched with branched arginine, exhibits excellent mRNA delivery properties and safety. These findings highlight its potential as a promising gene therapy tool.

OBJECTIVE: To evaluate the clinical application of indocyanine green (ICG) visualization by near infrared fluorescence laparoscopy (NIFL) in complex upper urinary tract reconstructions surgery. METHODS: This was a retrospective study of 7 patients who underwent complex surgeries of ureteral reconstruction between May 2019 and October 2019. There were 6 males and 1 female with the age ranging from 24 to 57 years (median age was 47 years). There were 5 cases of right ureteral strictures, of which 3 were proximal ureteral strictures and 2 were multiple and long ureteral strictures caused by radiotherapy. There were 2 cases of left ureteral strictures, of which 1 was ureteropelvic junction stricture and 1 was proximal ureteral stricture. There were 4 cases of secondary repair operations and 3 cases of primary operations. All the patients underwent laparoscopic surgery via the abdominal approach. ICG was injected into the ureter via nephrostomy tube during the operations, and the diseased ureter was identified by NIFL. Among the patients, 2 cases underwent IUPU (Institute of Urology, Peking University) modified ileal ureter replacement, 2 cases underwent ureteroureterostomy, 2 cases underwent appendiceal onlay flap ureteroplasty, and 1 case underwent lingual mucosa onlay flap ureteroplasty. RESULTS: All the operations were successfully completed without open conversion. The localization and separation of ureteral lesions were completed under NIFL. The mean operative time was 187 (135-300) min. The duration of ureteral separation was 15-27 min, and the mean time was 18 min. The estimated blood loss was 15-200 mL, the mean estimated blood loss was 50 mL. There was one patient with ileal ureter replacement who had fever after surgery and responded well to antibiotics. The mean (range) length of postoperative hospital stay was 7 (6-10) days and no postoperative complications of a high grade (Clavien-Dindo Ⅲ and Ⅳ) occurred. Up to now, the mean follow-up duration was 9 (6-11) months, and no indocyanine green toxicity occurred. All D-J stents and nephrostomy were removed successfully 2 months after the operation. Ultrasound showed no obvious hydronephrosis, and CTU (computed tomography urography) showed that the urinary tract was unobstructed and the kidney function was normal. CONCLUSION The application of ICG in the complex upper urinary tract reconstructive surgery is a safe and easy method to help surgeon to identify the ureter which may reduce the risk of iatrogenic damage and protect the ureteral blood supply.
Adult ; Female ; Humans ; Indocyanine Green ; Laparoscopy ; Male ; Middle Aged ; Retrospective Studies ; Ureter/surgery* ; Ureteral Obstruction ; Urologic Surgical Procedures ; Young Adult

OBJECTIVETo investigate the effect of tetrandrine (TET) on zinc finger protein 139 (ZNF139) and multidrug resistance (MDR) of human gastric carcinoma cell lines and possible mechanisms.

METHODSCultured SGC7901 and SGC7901/ADR were treated with TET (0.5, 1.0, 1.5, 2.0, and 2.5 microg/mL), then inhibition rates were measured by MTT assay in vitro. The expressions of ZNF139, MRP-1, MDR1, and GST-pi were detected by RT-PCR. The correlation between ZNF139 and each multidrug resistance factor was analyzed using Spearman correlation analysis, and the coefficient correlation was calculated.

RESULTSThe inhibition rate of TET (< or = 2.0 microg/mL) for SGC7901 and SGC7901/ADR was less than 10% with MTT assay. Expressions of ZNF139, MRP-1, MDR1, and GST-pi mRNA were higher in SGC7901/ADR than in SGC7901 (all P < 0.05). The expressions of ZNF139, MRP-1, MDR1, and GST--pi were down-regulated in SGC7901/ADR cells efficiently (all P < 0.01). Positive correlation existed between ZNF139 and MRP-1, ZNF139 and MDR1 before treated by TET in SGC7901/ADR, and this relationship also existed in SGC7901/ADR cells after treated by TET (all P < 0.05).

CONCLUSIONTET could achieve MDR reversion in gastric cancer cells by down-regulating the expression of ZNF139, MRP-1, and MDR1.

ATP Binding Cassette Transporter, Sub-Family B ; metabolism ; Benzylisoquinolines ; pharmacology ; Cell Line, Tumor ; Drug Resistance, Multiple ; drug effects ; genetics ; Drug Resistance, Neoplasm ; drug effects ; genetics ; Humans ; Kruppel-Like Transcription Factors ; metabolism ; Multidrug Resistance-Associated Proteins ; metabolism ; Stomach Neoplasms ; metabolism ; Zinc Fingers ; genetics

Objective To review the experience in diagnosis and treatment of pheochromocytoma in a single center. Methods A total number of 142/145 pheochromocytoma cases treated surgically in our institute from August 2002 to February 2010 were retrospectively reviewed. The mean diameter of tumor was 5.9 cm (1.3- 18. 0 cm). The majority of the tumors (92.9%) were adrenal pheochromocytomas. Ninety-eight patients (69.0 % ) presented initially with hypertension, whereas 44 patients (31%)presented with adrenal incidentaloma. A specific anti-hypertensive pre-surgery preparation with phenoxybenzamine or doxazosine mesylate was started over 1 week before the operation.Of the 142 patients, 91 accepted open surgery, 54 accepted laparoscopic surgery, of which, 5 converted from laparoscopic surgery to open surgery. Results Histopathological results showed that all the cases were pheochromocytoma, while 83 cases were benign, 23 cases were malignant and 37 cases were suspected malignant. Sudden rising of blood pressure during operation was related to the preoperative serum level of catecholamine. Eighty-seven of 98 patients with preoperative hypertension had normal postoperative blood pressure; the remaining 11 patients reduced the dosage of anti-hypertension medication postoperatively. During the follow-up of 3-96 months (median 46 months), 10 of 91patients had a recurrence or metastasis. Six patients died of recurrences or metastasis within 5 years.Conclusions The procedures of qualitative and locative diagnosis of phechromocytoma include clinical manifestations, biochemical tests and imaging investigation. Surgical excision is the fundamental treatment for cure. Patients with high serum level of catecholamine tend to have a sudden rising of blood pressure during operation. Preoperative management is extremely important for the safety of the patient. Intensive follow up is necessary.

OBJECTIVETo explore the value of flow cytometric immunophenotyping (FCI) in the diagnosis and differentiated diagnosis of lymphoma and explain the immunophenotypic features and differences of malignant lymphoma.

METHODSSeventy four fresh samples of suspicious lymphoma were collected from Nov. 2004 to Aug. 2006. Each sample was individually evaluated by FCI. The results were analyzed and compared with the histopathological diagnosis.

RESULTSAmong the 74 cases, the FCI data consisted with the final morphological diagnosis in 61 cases (82.4%). For the diagnosis of B and T non-Hodgkin's lymphoma (NHL), thymoma, carcinoma and benign lesions of lymph node, the concordance between FCI data and morphological diagnosis were 93.5%, 100%, 100%, 100% and 81.3%, respectively.

CONCLUSIONMulti-parameter FCI analysis can provide important information and help for diagnosis of lymphoma. It is an assistant but necessary approach for the diagnosis and differential diagnosis of lymphoma.

Adolescent ; Adult ; Aged ; Child ; Female ; Flow Cytometry ; Humans ; Immunophenotyping ; methods ; Lymphoma ; diagnosis ; immunology ; pathology ; Male ; Middle Aged