Twenty years of Gendicine® rAd-p53 cancer gene therapy: The first-in-class human cancer gene therapy in the era of personalized oncology.

Genetic mutations in TP53 contribute to human malignancies through various means. To date, there have been a variety of therapeutic strategies targeting p53, including gene therapy to restore normal p53 function, mutant p53 rescue, inhibiting the MDM2-p53 interaction, p53-based vaccines, and a number of other approaches. This review focuses on the functions of TP53 and discusses the aberrant roles of mutant p53 in various types of cancer. Recombinant human p53 adenovirus, trademarked as Gendicine, which is the first anti-tumor gene therapy drug, has made tremendous progress in cancer gene therapy. We herein discuss the biological mechanisms by which Gendicine exerts its effects and describe the clinical responses reported in clinical trials. Notably, the clinical studies suggest that the combination of Gendicine with chemotherapy and/or radiotherapy may produce more pronounced efficacy in slowing tumor growth and progression than gene therapy/chemotherapy alone. Finally, we summarize the methods of administration of recombinant human p53 adenovirus for different cancer types to provide a reference for future clinical trials.

A positive feedback cycle between the alarmin S100A8/A9 and NLRP3 inflammasome-GSDMD signalling reinforces the innate immune response in Candida albicans keratitis.

OBJECTIVE: Fungal keratitis is a severe sight-threatening ocular infection, without effective treatment strategies available now. Calprotectin S100A8/A9 has recently attracted great attention as a critical alarmin modulating the innate immune response against microbial challenges. However, the unique role of S100A8/A9 in fungal keratitis is poorly understood. METHODS: Experimental fungal keratitis was established in wild-type and gene knockout (TLR4-/- and GSDMD-/-) mice by infecting mouse corneas with Candida albicans. The degree of mouse cornea injuries was evaluated by clinical scoring. To interrogate the molecular mechanism in vitro, macrophage RAW264.7 cell line was challenged with Candida albicans or recombinant S100A8/A9 protein. Label-free quantitative proteomics, quantitative real-time PCR, Western blotting, and immunohistochemistry were conducted in this research. RESULTS: Herein, we characterized the proteome of mouse corneas infected with Candida albicans and found that S100A8/A9 was robustly expressed at the early stage of the disease. S100A8/A9 significantly enhanced disease progression by promoting NLRP3 inflammasome activation and Caspase-1 maturation, accompanied by increased accumulation of macrophages in infected corneas. In response to Candida albicans infection, toll-like receptor 4 (TLR4) sensed extracellular S100A8/A9 and acted as a bridge between S100A8/A9 and NLRP3 inflammasome activation in mouse corneas. Furthermore, the deletion of TLR4 resulted in noticeable improvement in fungal keratitis. Remarkably, NLRP3/GSDMD-mediated macrophage pyroptosis in turn facilitates S100A8/A9 secretion during Candida albicans keratitis, thus forming a positive feedback cycle that amplifies the proinflammatory response in corneas. CONCLUSIONS: The present study is the first to reveal the critical roles of the alarmin S100A8/A9 in the immunopathology of Candida albicans keratitis, highlighting a promising approach for therapeutic intervention in the future.

Wire Electrochemical Micromachining of Aluminum Rings for the Fabrication of Short-Millimeter Corrugated Horns.

With the increase of working frequency, the feature size of a corrugated horn will be greatly reduced, causing challenges for fabrication. This paper investigated wire electrochemical micromachining (WECMM) of aluminum rings for assembly of a mandrel for electroforming, which has been a primary method for producing corrugated horns. By utilizing a rotary helical electrode and green additives, the removal efficiency of electrolytic products in WECMM was improved. It was found that the machined slits had good unilateral consistency on the left side of the electrode feeding direction when the electrode rotated clockwise. Complexing agent glutamic diacetic acid (GLDA) can compete with OH- for Al3+ and has an obvious effect in reducing insoluble electrolytic products. From experimental investigations on typical parameters, an optimal parameter combination considering slit homogeneity and machining efficiency was obtained. In an electrolyte solution containing 15 g/L sodium nitrate solution and 15 g/L GLDA, 100 µm-thick aluminum rings with good edge and surface qualities were fabricated at a rate of 1.2 µm/s using a helical electrode with a diameter of 0.3 mm. Finally, these aluminum rings were successfully applied to make an internal corrugated sample with a rib width of 100 µm and a groove depth of 500 µm.

Targeting neuronal and glial cell types with synthetic promoter AAVs in mice, non-human primates and humans.

Targeting genes to specific neuronal or glial cell types is valuable for both understanding and repairing brain circuits. Adeno-associated viruses (AAVs) are frequently used for gene delivery, but targeting expression to specific cell types is an unsolved problem. We created a library of 230 AAVs, each with a different synthetic promoter designed using four independent strategies. We show that a number of these AAVs specifically target expression to neuronal and glial cell types in the mouse and non-human primate retina in vivo and in the human retina in vitro. We demonstrate applications for recording and stimulation, as well as the intersectional and combinatorial labeling of cell types. These resources and approaches allow economic, fast and efficient cell-type targeting in a variety of species, both for fundamental science and for gene therapy.

The differences of cell wall in roots between two contrasting soybean cultivars exposed to cadmium at young seedlings.

The plant root cell wall (CW) is the first structure that comes into contact with extracellular cadmium (Cd), and it plays an important role in the absorption, immobilization, and translocation of Cd in the roots. However, the differences in the cell wall components between Cd-tolerant and Cd-sensitive cultivars are unclear. A hydroponic experiment was carried out to investigate the differences in the concentrations of Cd, total sugars, and uronic acid in pectin, hemicellulose 1, hemicellulose 2, cellulose, and lignin, as well as pectin methylesterase enzyme activity (PME) in the roots of two soybean cultivars that differ with respect to Cd tolerance exposed to 0 and 23 µM Cd treatments. The bound forms of Cd in the roots were found to differ between the two soybean genotypes; 50.2% of the Cd in the root cell wall accumulates in the pectin in the highly Cd-tolerant and low Cd-accumulating cultivar HX3, while 50.6% of the root cell wall Cd accumulates in cellulose in the Cd-sensitive and high Cd-accumulating cultivar BX10. The total sugar and uronic acid concentrations of the cell wall components increased in response to Cd stress, while the concentrations of total sugars and uronic acid in BX10 were higher than in HX3 (except for hemicellulose 1). Increased demethylation of pectin may be the main reason that Cd is mainly concentrated in the primary wall in HX3, because the PME activity was higher in HX3 than it was in BX10 under Cd treatment. Furthermore, BX10 had a higher lignin concentration after Cd treatment, and showed the same change in cellulose. Cd in the root cell wall of BX10 was fixed in the secondary cell wall, which may be a result of the coupling to cellulose and lignin. In conclusion, root cell walls in soybean cultivars that differ in Cd tolerance may possess different mechanisms to prevent Cd from entering cells, and the sequestration of Cd in different cell wall components may determine the differences in Cd tolerance between the two genotypes.

Genome-wide characterization of soybean P 1B -ATPases gene family provides functional implications in cadmium responses.

BACKGROUND: The P1B-ATPase subfamily is an important group involved in transporting heavy metals and has been extensively studied in model plants, such as Arabidopsis thaliana and Oryza sativa. Emerging evidence indicates that one homolog in Glycine max is also involved in cadmium (Cd) stress, but the gene family has not been fully investigated in soybean. RESULTS: Here, we identified 20 heavy metal ATPase (HMA) family members in the soybean genome, presented as 10 paralogous pairs, which is significantly greater than the number in Arabidopsis or rice, and was likely caused by the latest whole genome duplication event in soybean. A phylogenetic analysis divided the 20 members into six groups, each having conserved or divergent gene structures and protein motif patterns. The integration of RNA-sequencing and qRT-PCR data from multiple tissues provided an overall expression pattern for the HMA family in soybean. Further comparisons of expression patterns and the single nucleotide polymorphism distribution between paralogous pairs suggested functional conservation and the divergence of HMA genes during soybean evolution. Finally, analyses of the HMAs expressed in response to Cd stress provided evidence on how plants manage Cd tolerance, at least in the two contrasting soybean genotypes examined. CONCLUSIONS: The genome-wide identification, chromosomal distribution, gene structures, and evolutionary and expression analyses of the 20 HMA genes in soybean provide an overall insight into their potential involvement in Cd responses. These results will facilitate further research on the HMA gene family, and their conserved and divergent biological functions in soybean.

Root morphological responses of five soybean [Glycine max (L.) Merr] cultivars to cadmium stress at young seedlings.

To examine the differences in root morphological responses of soybean cultivars with different cadmium (Cd) tolerance and accumulation to Cd stress, the biomass, Cd concentration, and root morphological features of five soybean cultivars were determined under 0, 9, 23, 45, and 90 µM Cd stress via hydroponic experiments. Significantly genotypic differences in Cd tolerance and Cd concentration were observed between five soybean cultivars at four Cd levels. For Cd tolerance, HX3 showed a strong Cd tolerance with tolerance indexes of shoot biomass at 92.49, 76.44, 60.21, and 46.45% after 18 days at four Cd levels, and others had similar weak tolerance at young seedling. For Cd accumulation, Cd concentration in roots showed far higher than that in shoots. The different accumulation features in roots and shoots among five cultivars were found at four Cd levels. Comparing with the control, the total root length (RL), root surface area (SA), and root volume (RV) of all cultivars were decreased significantly at four Cd levels. Tolerant cultivar HX3 had the largest root system and sensitive cultivar BX10 had the smallest root system at young seedling stage. Correlation analysis indicated that RL, SA, and RV were positively correlated with root biomass and shoot biomass under 9 and 23 µM Cd treatments, but root average diameter (RD) was negatively correlated with shoot biomass and root biomass only under 9 µM Cd treatments, while RL and SA were negatively correlated with root Cd concentration under 23 and 45 µM Cd treatments. The results suggested that root morphological traits were closely related to Cd tolerance at young seedlings under Cd treatments.

Comparison of subcellular distribution and chemical forms of cadmium among four soybean cultivars at young seedlings.

The hydroponic experiment was carried out to investigate the Cd subcellular distribution and chemical forms in roots and shoots among four soybean seedling cultivars with two Cd treatments. HX3 and GC8, two tolerant and low-grain-Cd-accumulating cultivars, had the lowest Cd concentration in roots and high Cd concentration in shoots, while BX10 and ZH24, two sensitive and high-grain-Cd-accumulating cultivars, had the highest Cd concentration in roots and the lowest Cd concentration in shoots at young seedling stage. Furthermore, the sequence of Cd subcellular distribution in roots at two Cd levels was cell wall (53.4-75.5 %) > soluble fraction (15.8-40.4 %) > organelle fraction (2.0-14.7 %), but in shoots, was soluble fraction (39.3-74.8 %) > cell wall (16.0-52.0 %) > organelle (4.8-19.5 %). BX10 and ZH24 had higher Cd concentration in all subcellular fractions in roots, but HX3 and GC8 had higher Cd concentration of soluble fraction in shoots. The sequence of Cd chemical forms in roots was FNacl (64.1-79.5 %) > FHAC (3.4-21.5 %) > Fd-H2O (3.6-13.0 %) > Fethanol (1.4-21.8) > FHCl (0.3-1.6 %) > Fother (0.2-1.4 %) at two Cd levels but, in shoots, was FNacl (19.7-51.4 %) ≥ FHAC (10.2-31.4 %) ≥ Fd-H2O (8.8-28.2 %) ≥ Fethanol (8.9-38.6 %) > FHCl (0.2-9.6 %) > Fother (2.5-11.2 %). BX10 and ZH24 had higher Cd concentrations in each extracted solutions from roots, but from shoots for GC8 and HX3. Taken together, the results uncover that root cell walls and leaf vacuoles might play important roles in Cd detoxification and limiting the symplastic movement of Cd.

Laparoendoscopic single-site ureteroureterostomy with intraoperative retrograde ureteroscopy-assisted technique for benign proximal and middle ureteral strictures: a single-center experience.

OBJECTIVE: To report a minimally invasive and reproducible technique that greatly facilitates the identification of the stricture during laparoendoscopic single-site ureteroureterostomy (LESS-UU) for benign proximal and middle ureteral strictures, using the intraoperative retrograde ureteroscopy-assisted technique. PATIENTS AND METHODS: Between April 2011 and January 2013, 13 patients with a benign proximal or middle ureteral stricture underwent LESS-UU at our institution. A combination of diuretic renal scans, antegrade/retrograde ureteropyelography and/or computed tomography, and stent placement or exchange was preoperatively performed to assess all patients. The intraoperative retrograde ureteroscopy-assisted technique was used to identify the exact position of the stricture and place the stenting during LESS-UU. RESULTS: Intraoperative retrograde ureteroscopy was successfully performed in all cases. The mean operative time was 156 minutes (range, 125-190 minutes), and the estimated blood loss was 80 mL (range, 20-160 mL). The mean hospital stay was 5 days (range, 4-7 days). One patient required conversion to open surgery because of the severe adhesions surrounding the stricture that resulted in failure to progress. Urine leakage occurred in 1 patient postoperatively and was successfully treated by conservative management. Postoperative fever occurred in another patient, who was treated with a dose of oral antibiotics. No major intraoperative or postoperative complication occurred. Clinical and radiographic success was achieved in 100% (13/13) of patients during a mean follow-up of 13.1 months (range, 9-27 months). CONCLUSIONS: LESS-UU is feasible and safe for repairing benign proximal and middle ureteral stricture. The intraoperative retrograde ureteroscopy-assisted technique during LESS-UU is useful for localizing the stricture.

Retroperitoneoscopic upper pole nephroureterectomy in duplex kidney: focus on the role of dilated upper pole ureter.

PURPOSE: To describe our technique and experience with retroperitoneoscopic upper pole nephroureterectomy in duplex kidney, focusing on the role of dilated upper ureter. MATERIALS AND METHODS: From November 2004 to August 2011, retroperitoneoscopic upper pole nephroureterectomy was performed in 31 patients with a duplex kidney by a single, experienced laparoscopic surgeon. We developed our own surgical technique to suit this technically challenging procedure. Follow-up studies were performed using renal ultrasonography, intravenous urography (IVU) and/or dimercaptosuccinic acid (DMSA) renal scan in all patients at 3 months postoperatively and annually thereafter. RESULTS: All procedures were completed laparoscopically without conversion to open surgery and blood transfusion. The mean operative time was 106 (90-157) min. The estimated blood loss was < 50 mL in all cases. The mean postoperative hospital stay was 4.2 (3-7) days. Perioperative complications were limited to 1 case of peritoneal tear during a procedure and 1 case of transient postoperative fever. No major intraoperative and postoperative complication occurred. With the mean follow-up period of 41 months (range 3 to 80), no case was observed to have functional loss of the remaining lower moiety on postoperative IVU or DMSA renal scan. CONCLUSION: Retroperitoneoscopic upper pole nephroureterectomy using our technique is safe and effective.

Laparoendoscopic single-site retroperitoneoscopic adrenalectomy for pheochromocytoma: case selection, surgical technique, and short-term outcome.

PURPOSE: To present our experience with case selection and operative skills of laparoendoscopic single-site (LESS) retroperitoneoscopic adrenalectomy for pheochromocytoma and evaluate its feasibility. PATIENTS AND METHODS: Between June 2011 and December 2012, we performed LESS retroperitoneoscopic adrenalectomy for 16 patients with pheochromocytoma. In all patients, the diameter of the pheochromocytoma was less than 4.0 cm. During the operation, a single-port access was inserted through a 2.5-3.0 cm transverse incision below the tip of the 12th rib. Internally, the operative procedure duplicates the conventional retroperitoneoscopic adrenalectomy for pheochromocytoma. RESULTS: No conversions to open surgery or standard laparoscopy with additional trocars were necessary. The mean operative duration was 68.1 minutes (range 41-125 min). The mean blood loss was negligible (<50 mL), and no patient needed blood transfusion. Intraoperative hypertension (SBP>180 mmHg) occurred in 12.5% (2/16) of the patients. No patient had sustained hypertension, and none experienced intraoperative hypotension (systolic blood pressure <80 mm Hg). The only postoperative complication was one case of pneumonia successfully treated with antibiotics. The average postoperative hospital stay was 3.1 days (range 2-5 days). All patients left the hospital with a good cosmetic appearance. CONCLUSIONS: In properly selected patients, LESS retroperitoneoscopic adrenalectomy is a feasible and safe procedure for pheochromocytoma.

Identification and comparative analysis of cadmium tolerance-associated miRNAs and their targets in two soybean genotypes.

MicroRNAs (miRNAs) play crucial roles in regulating the expression of various stress responses genes in plants. To investigate soybean (Glycine max) miRNAs involved in the response to cadmium (Cd), microarrays containing 953 unique miRNA probes were employed to identify differences in the expression patterns of the miRNAs between different genotypes, Huaxia3 (HX3, Cd-tolerant) and Zhonghuang24 (ZH24, Cd-sensitive). Twenty six Cd-responsive miRNAs were identified in total. Among them, nine were detected in both cultivars, while five were expressed only in HX3 and 12 were only in ZH24. The expression of 16 miRNAs was tested by qRT-PCR and most of the identified miRNAs were found to have similar expression patterns with microarray. Three hundred and seventy six target genes were identified for 204 miRNAs from a mixture degradome library, which was constructed from the root of HX3 and ZH24 with or without Cd treatment. Fifty five genes were identified to be cleaved by 14 Cd-responsive miRNAs. Gene ontology (GO) annotations showed that these target transcripts are implicated in a broad range of biological processes. In addition, the expression patterns of ten target genes were validated by qRT-PCR. The characterization of the miRNAs and the associated target genes in response to Cd exposure provides a framework for understanding the molecular mechanism of heavy metal tolerance in plants.

Transperitoneal mini-laparoscopic pyeloplasty and concomitant ureteroscopy-assisted pyelolithotomy for ureteropelvic junction obstruction complicated by renal caliceal stones.

OBJECTIVE: To present our experience of combining transperitoneal mini-laparoscopic pyeloplasty (mini-LP) and concomitant ureteroscopy-assisted pyelolithotomy (U-P) for ureteropelvic junction obstruction (UPJO) complicated by renal caliceal stones in the same session. METHODS: Between May 2007 and December 2011, mini-LP and concomitant U-P was performed in nine patients with UPJO and ipsilateral renal caliceal stones. Stone location and burden were preoperatively assessed. After pyelotomy with appropriate length (about 4 mm), a 16-Fr catheter sheath replaced the uppermost or lowermost laparoscopic trocar and was introduced directly into the renal pelvis under the guidance of a guide wire and laparoscopic vision. A 7.5F rigid ureteroscopy passed through the catheter sheath into the plevis. Intracorporeal lithotripsy and/or pressure irrigation via a pump was used for caliceal stone removal. Subsequently, laparoscopic pyeloplasty was performed in a standard fashion. Postoperative imaging was assessed. RESULTS: The calculi sizes ranged from 2 to 11 mm (mean, 7.1 mm) and an average of 3 stones per patient was removed (range, 1 to 6 stones). Complete stone clearance confirmed by postoperative imaging was achieved in all patients. Mean operative time was 210 minutes, and estimated blood loss was 20 mL. Mean hospital stay was 5 days (4-7). Stent was removed after 4-8 weeks. No intraoperative or postoperative complications were noted during a mean follow-up of 18.5 months (range, 6 to 24 months). CONCLUSIONS: Mini-LP and concomitant U-P are simple and effective alternatives for the simultaneous management of UPJO complicated by coexisting ipsilateral renal caliceal stones.

Retroperitoneal laparoendoscopic single-site ureterolithotomy and ureteroureterostomy for retrocaval ureter with ureteral calculus: first case report.

A 53-year-old man presented with a 6-month history of intermittent right flank pain. Radiological imaging confirmed the diagnosis of retrocaval ureter (RCU) and ureteral calculus. Retroperitoneal laparoendoscopic single-site surgery (LESS) ureterolithotomy and ureteroureterostomy was successfully performed. The operative time was 185 min and the blood loss was approximately 20 ml. The patient's postoperative course was uneventful. Postoperative analgesia was not needed. The patient was discharged on the third postoperative day. The drain and double-J stent were respectively removed at 1 and 8 weeks postoperatively. At the 3-month follow-up, nuclear scan showed no evidence of obstruction of the right kidney and the patient also remained symptom free. It may be concluded that retroperitoneal LESS repair for RCU is a feasible and safe procedure, which can be considered as a option for the management of RCU even if it is complicated by the presence of a ureteral calculus.