BACKGROUND: Studies regarding Feridex in vitro cell labeling are mainly in rodents, while little information is known on primate crab-eating macaque.OBJECTIVE: To explore the feasibility of protocols using Feridex and transfection agents for in vitro magnetic labeling of bone marrow stromal cells (BMSCs) in crab-eating macaque.METHODS: Under the sterile condition, the crab-eating macaque BMSCs were obtained by means of density gradient centrifugation following a bone puncture. Feridex-poly-l-lysine complexes were used to magnetically label BMSCs. The efficiency and cellular viability of Feridex-poly-l-lysine labeled BMSCs were evaluated by Prussian blue staining, electron microscopy, and trypan blue dye exclusion test. The proliferation and differentiation ability of Feridex-poly-l-lysine labeling BMSCs were also investigated by inverted phase contrast microscope and immunocytochemistry. RESULTS AND CONCLUSION: BMSCs could be effectively labeled by Feridex and labeling efficiency was around 99%. Tiny blue stained fine particles and numerous vesicles coated with the electron-dense magnetic iron particles could be found in the cytoplasm of Feridex-poly-l-lysine labeled BMSCs under optical microscopy and transmission electron microscopy respectively. Cell viability, proliferation and differentiation ability of labeled BMSCs were not affected by Feridex-poly-l-lysine labeling. Results indicated that Feridex might be used to label BMSCs of crab-eating macaque.

Objective: To detect the mutation of epidermal growth factor receptor (EGFR) gene in peripheral blood of non-small cell lung cancer (NSCLC) patients in Yunnan area with Super-ARMS, and to explore its correlation with clinicopathological characteristics. Methods: A total of 222 blood samples from patients with NSCLC were collected between January 2017 to December 2018 in the Molecular Diagnostic Center of Yunnan Cancer Hospital. The EGFR gene mutation in peripheral blood samples was detected by SuperARMS, and the relationship between EGFR gene mutation and clinicopathological features was analyzed. Meanwhile, the independent risk factors influencing EFGR mutation were also analyzed. Results: In the peripheral blood of 222 NSCLC patients, there were 81 cases (36.5%) with EGFR gene mutation. Among them, exon 19 deletion and L858R gene point mutation were the most common (75.3% of total mutation); female patients had a higher mutation rate than male patients (45.9% vs 27.0%); patients <60 years old had a higher incidence of mutation than patients≥60 years old (43.2% vs 28.8%) (P<0.05 or P<0.01); moreover, patients with no history of smoking, no history of radical surgery, adenocarcinoma, advanced stage and no history of chemotherapy had higher incidence of EGFR mutation (43.9% vs 21.6%, 39.2% vs 21.2%, 43.9% vs 4.8%, 39.7% vs 23.3% and 44.0% vs 23.5%) (P<0.05 or P<0.01). Multivariate logistic analysis showed that young, no smoking history, adenocarcinoma and no surgical history were independent risk factors for EGFR gene mutation (all P<0.01). Conclusion: In the peripheral blood of patients with NSCLC in Yunnan, the mutation rate of EGFR gene is higher in patients with age<60 years old, adenocarcinoma and non-smoking. Super-ARMS method is more sensitive in the detection of EGFR mutation in peripheral blood of lung cancer patients.

Objective To discuss the feasibility of laparoscopic simple oblique side-to-side duodenoduodenostomy in treating congenital annular pancreas in neonates.Methods Medical records of neonates with the diagnosis of congenital annular pancreas undergoing laparoscopic simple oblique side-to-side duodenoduodenostomy at Huai'an Women and Children's Hospital from January 2015 to May 2017 were reviewed.The age ranged from 1 d to 4d with a median age of 2d at operation.The body weight at operation was 2.5-3.5 kg with a mean of 3.1 kg.By a lower-pressure pneumoperitoneum of 5-8 mmHg (1 mmHg =0.133 kPa) and a suspending suture for liver elevator,the procedure was performed using 4 ports.A transumbilical 5-mm port was for the laparoscopy and 3 3-mm trocars were placed for inserting instruments.The diagnosis was made after laparoscopic exploration,and then the distal duodenum was incised longitudinally 0.5-1 cm away from the annular pancreas,the proximal duodenum was incised O.5 cm away from the annular pancreas obliquely.The duodenoduodenostomy was performed as a "simple oblique" side-to-side anastomosis with 5-0 PDS running suture.Results All the 6 cases were treated by laparoscopic simple oblique side-to-side duodenoduodenostomy.Three of cases were annular pancreas complicated with congenital intestinal malrotation,and Ladd's procedure was accomplished in the mean time.The average operative time was 98 min (70-120 min).The blood loss during operation was 3-10 ml.Fluid diet started on postoperative day 3 to 7 (mean 4.5 d),without abdominal distention and vomit,and all the cases were discharged uneventfully in a median of 7-14 (mean 9 d) postoperative day without any postoperative complications.No cases were transferred to open abdominal surgery,and there were no intraoperative and postoperatuve adverse events.The cases were followed-up for 1-30 months (mean 13 months),and all the cases grew well without intestinal obstruction.Conclusions The laparoscopic simple oblique duodenoduodenostomy in treating congenital annular pancreas was conveniently performed,the intestinal function recovered quickly,which can be safely done in neonatal period even if the neonate was complicated with congenital intestinal malrotation.

Objective:To explore the possibility of laparoscopic simple oblique duodenoduodenostomy in the management of congenital duodenal obstruction in neonates.Methods:The clinical data of neonates with congenital duodenal obstruction undergoing laparoscopic simple oblique duodenoduodenostomy at Huaian Women and Children′s Hospital from February 2015 to February 2018 were retrospectively reviewed.By a lower-pressure pneumoperitoneum of 5-8mmHg and a suspending suture for right liver elevator, the procedure was performed using 4 ports: a transumbilical 5-mm port for the camera and another three 3-mm ports for instruments.After kocherizing of the proximal and distant part of the duodenum, the lower duodenum was incised longitudinally 0.5-1cm distal to the blocked end, the upper duodenum incision was placed 0.5cm away from the blocked end extended downward obliquely.The duodenoduodenostomy was performed as a " simple" " oblique" anastomosis with 5-0 PDS suture.Results:Fifteen patients were identified with a median age at operation of 2 days.Of all the cases, 4 cases were duodenal atresia (type Ⅲ), 11 cases were annular pancreas, and 7 cases of them were annular pancreas complicated with congenital intestinal malrotation.All cases were treated with laparoscopic simple oblique duodenoduodenostomy.Ladd procedure was accomplished in 7 cases with congenital intestinal malrotation in the meantime.The average operative time was (115.0±25.5) min (70-145 min). Feedings were started on postoperative 4-10d (mean 6.0d), without abdominal distention and vomit, all patients were discharged in good health with a median postoperative hospital stay of 7-14d(mean 10.6d). There was no mortality rate, no conversion to laparotomy, and no intraopertive complications in the group.The cases were followed-up for 2-36 months (mean 15.6 months), and all cases were doing well.Conclusion:Laparoscopic simple oblique duodenoduodenostomy is safe and efficacious in neonatal period, and can be a viable option in the treatment of neonates with congenital duodenal obstruction.

Objective To explore the clinical value of portal exposure in laparoscopic treatment of children with type Ⅲ biliary atresia (BA).Methods From June 2013 to October 2017,30 infants with type Ⅲ BA who treated with laparoscopic portoenterostomy in Huai'an Women and Children's Hospital were selected.A percutaneous suture was used to snare the round ligament and retract the liver,other percutaneous stay sutures were then introduced and fundus and neck of gallbladder were sutured to elevate the liver to expose the portal hepatis.The fibro cord and hepatic vessels were mobilized,and then two rubber bands were put around the left and right portal veins and hepatic arteries.The portal hepatis was exposed by laterally stretching the two elastic rubber bands.The fibro cord was removed and then laparoscopic portoenterostomy was accomplished.In 20 cases,the liver was enlarged,part of hepatic lobus quadratus was removed laparoscopiclly for exposure of the portal hepatis.Results There were 30 cases in this group,2 cases were converted to open surgery by a micro transverse incision.There was no surgical death.Time of laparoscopic procedure varied from 210 to 280 min.All cases survived the surgery without any intraoperative complications.Blood loss during operation was minimal,without necessity for blood transfusion.One case died of respiratory failure one week after surgery.Two cases were lost follow-up.Twenty-five cases were followed up for 3～51 months(mean 22.4 months).Three cases died because of repeated cholangitis and liver failure at 10,16,35 months postoperatively.Nineteen patients' total bilirubin had dropped to normal,three others' bilirubin levels dropped significantly after surgery.Conclusion The technique of laparoscopic hepatic porta exposure can help to complete hepatic portoenterostomy successfully,reduce the conversion rate of laparoscopic surgery,and improve the surgical effect.

Objective To investigate epidermal growth factor receptor(EGFR)gene T790M mutation in plasmatic ctDNA samples from 171 patients with non-small cell lung cancer and analyze the relationship between EGFR T790M mutation and the clinical factors. Methods The EGFR T790M mutation was detected in 171 cases by super amplification refractory mutation system(Super ARMS)in this paper. Rusults The EGFR gene T790M mutation was identified in 7.60%(13/171)plasmatic ctDNA samples which mostly came from patients withⅢb~Ⅳstages of lung cancer. The EGFR T790M mutation rate was identified in 2.05%(3/146)plasmatic samples of pa-tients who did not received treatment of EGFR-TKIs,which was lower than 40.00%(10/25,P<0.05)plasmatic samples of patients who received treatment of first generational EGFR-TKIs. The EGFR T790M mutation rate was identified in 75.00%(3/4) and 60.00%(6/10) plasmatic samples of patients who have received TKI for 6 to 10 months and more than 10 months,which was higher than 9.10%(1/11,P < 0.05)plasmatic samples of patients who have received TKIs for less than 6 months. Conclusions This article demonstrated that EGFRT790M muta-tion was more common in lately NSCLC patients who have received TKIs treatmentover 6 months,meanwhile the EGFR T790M mutation dynamical detective technology will effectively guide the clinic treatment.

Near-infrared (NIR)-light-triggered nanomedicine, including photodynamic therapy (PDT) and photothermal therapy (PTT), is growing an attractive approach for cancer therapy due to its high spatiotemporal controllability and minimal invasion, but the tumor eradication is limited by the intrinsic anti-stress response of tumor cells. Herein, we fabricate a tumor-microenvironment responsive CRISPR nanoplatform based on oxygen-deficient titania (TiO_2-x ) for mild NIR-phototherapy. In tumor microenvironment, the overexpressed hyaluronidase (HAase) and glutathione (GSH) can readily destroy hyaluronic acid (HA) and disulfide bond and releases the Cas9/sgRNA from TiO_2-x to target the stress alleviating regulators, i.e., nuclear factor E2-related factor 2 (NRF2) and heat shock protein 90α (HSP90α), thereby reducing the stress tolerance of tumor cells. Under subsequent NIR light illumination, the TiO_2-x demonstrates a higher anticancer effect both in vitro and in vivo. This strategy not only provides a promising modality to kills cancer cells in a minimal side-effects manner by interrupting anti-stress pathways but also proposes a general approach to achieve controllable gene editing in tumor region without unwanted genetic mutation in normal environments.

[This corrects the article DOI: 10.1016/j.apsb.2022.06.016.].