Objective To predict the spatial distribution of Himalayan marmot using geographic information system (GIS) and Genetic Algorithm for Rule-set Production (GARP) model based on the monitoring data of Himalayan marmot in Qinghai Province.Methods Based on the data of routine monitoring and field survey by Global Position System (GPS) of Himalayan marmot,the position data was processed by spatial mapping using ArcGIS software.Ecological environment variables related to Himalayan marmot including terrain,slope,temperature,precipitation,vegetation,land use and other related variables were extracted and analyzed.The prediction model of Himalayan marmot distribution was constructed based on GARP model and ArcGIS software,and the spatial distribution mapping and analysis were carried out.Results Totally 198 points of Himalayan marmot were obtained by combining the recovery position with GPS information.The average model error of omission was 1.998 through the GARP modeling,while the optimal 100 model were highly statistically significant (all x2 ＞ 163.03,all P ＜ 0.01).The spatial distribution of predicted probability was divided into three grades including less than ＜ 40％,40％-＜ 80％ and 80％-100％ using ArcGIS,and the area with the prediction probability of 80％-100％ was the most suitable distribution area of Himalayan marmot.Conclusions The spatial distribution of plague host animal is predicted successfully using GIS and GARP ecological niche model.The result is more accurate compared to the statistic area by administrative region,which can provide important reference for plague prevention and control.

Objective: To construct the sponge vector which can target microRNA-186 (miR-186), and to create a stable EA. hy926 cell line that can knockdown miR-186. Methods: The miR-186 sponge sequence was chemically synthesized and cloned into lentiviral vector FV040. Then the FV040-miR-186-sponge recombinant plasmid together with the helper plasmids were cotransfected into the HEK293T cells by using lipofectamine 2000 to package lentivirus and the viral titer was determined. The FV040-control lentivirus (designated as the control group), and the FV040-miR-186-sponge (designated as the experiment group) were used to infect EA. hy926 cells for establishing stable cell lines. The fluorescent quantitative PCR (qPCR) method was used to detect the relative expression levels of miR-186 in the EA. hy926 cells in blank group, FV040-control group and FV040-miR-186 sponge group, respectively. Results: The cloned target sequence was identical with the designed miR-186 sponge sequence. The green fluorescence protein (GFP) was observed in the infected EA. hy926 cells. The lentivirus titre of viruses in the FV040-control group was 2×108 TU middot; mL-1, and which was 6 × 108 TU middot; mL-1 in FV040-miR- 186 sponge group. The EA. hy926 cell line stably expressed miR-186-sponge was established successfully and the infection rate was as high as 95%. The qPCR results indicated that the relative expression level of miR-186 in the EA. hy926 cells in FV040-miR-186-sponge group was lower than those in blank control group and FV040-control group (P<0. 01). Conclusion: The miR-186-sponge vector is successfully constructed, and the EA. hy926-miR- 186-sponge cell line with the stably decreased expression of miR-186 is established successfully.

Background:Osteosarcoma is the most common bone malignancy in children and adolescents, and 20%–30% of the patients suffer from poor prognosis because of individual chemoresistance. The Hippo/yes?associated protein (YAP) signaling pathway has been shown to play a role in tumor chemoresistance, but no previous report has focused on its involvement in osteosarcoma chemoresistance. This study aimed to investigate the role of the Hippo/YAP sign?aling pathway in osteosarcoma chemoresistance and to determine potential treatment targets.
Methods:Using the Cell Titer?Glo Luminescent cell viability assay and lfow cytometry analysis, we determined the proliferation and chemosensitivity of YAP?overexpressing and YAP?knockdown osteosarcoma cells. In addition, using western blotting and the real?time polymerase chain reaction technique, we investigated the alteration of the Hippo/YAP signaling pathway in osteosarcoma cells treated with chemotherapeutic agents.
Results:Mammalian sterile 20?like kinase 1 (MST1) degradation was increased, and large tumor suppressor kinase 1/2 (LATS1/2) total protein levels were decreased by methotrexate and doxorubicin, which increased activation and nuclear translocation of YAP. Moreover, YAP increased the proliferation and chemoresistance of MG63 cells.
Conclusions:The Hippo/YAP signaling pathway plays a role in osteosarcoma chemoresistance, and YAP is a potential target for reducing chemoresistance.

Objective:To explore the initial experience of thoracoscopic repair with simplified mattress sutures in the treatment of diaphragmatic hernia in neonates without posterolateral rim of diaphragm.Methods:A retrospective review of the new simplified technique in 10 cases from March 2015 to October 2017 was performed.Of the patients, 6 cases were male, 4 cases were female.The age was 10min-1d, 7 cases were term newborns, and 3 cases were premature.The mean weight was 2.88 kg(ranged 2.3-3.5kg). All the 4 cases were left-sided.Two to three primary suture sites were taken from the relative intercostal region of the body surface projection of the defect.A snip incision about 1 mm of the skin was done.Two 2-0 non-absorbable sutures round the rib were inserted between the front edged of the defect and the diaphragm muscle through a syringe needle.The first thread was brought out of the body by the ring of the second thread and knot tying was made extracorporally.The posterolateral defect was closed; the knot was under the skin of intercostals space.Results:Ten neonates with CDH were repaired successfully using this new simplified technique.The mean operative time was 37.5min(ranged 25-60min) for each CDH repair.No cases required conversion to open surgery, blood loss was minimal.The 10 cases were followed up for 16.5 months(ranged 5-24 months), with no death and no recurrence.One neonate complicated with subcutaneous emphysema postoperatively and healed in one week.Conclusion:The new technique of thoracoscopic repairing with simplified mattress sutures when no posterolateral rim of diaphragm exists has all the advantages of thoracoscopy in neonates combined with the advantages of reduced operative time, simplicity, feasibility and definite curative effect and has the value of clinical popularization.

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 explore the application of Body Tom® mobile CT combined with basic anesthesia in preoperative painless positioning of small pulmonary nodules, and evaluate its safety and effectiveness. Methods     Patients using mobile Body Tom® CT to accurately locate pulmonary nodules in the Department of Thoracic Surgery of Affiliated Nanjing Brain Hospital, Nanjing Medical University from August to October 2022 were retrospectively included. Clinical data of the whole patient group were analyzed. Results    We finally included 30 patients with 12 males and 18 females at age of 23-71 years. The position success rate of 30 patients with small pulmonary nodules was 100.0%. Location time was 14.20±4.07 min. There was one patient of intrapulmonary hemorrhage, with no other complications such as pneumothorax, positioning needle shedding, or pleural reaction. The time from the end of positioning to the start of surgery was 12.63±5.68 min. There was no needle migration or indocyanine green overflow. All patients completed resection of small pulmonary nodules under single-port thoracoscopy, no transit to opening chest. The average operation time was 85.32±12.60 min. There was no postoperative complications, and the average postoperative chest tube retention time was 2.12±1.34 days. And the average length of hospital stay was 3.52±1.45 days. The postoperative pathological results showed that the distance from the nodules was greater than 2 cm. Conclusion     Body Tom® mobile CT combined with basic anesthesia can achieve the preoperative painless, precise positioning of pulmonary nodules, effectively reduce the incidence of preoperative positioning complications, shorten the operation waiting time, ensure the safety and effectiveness of patients with preoperative pulmonary nodules positioning, and further improve the surgical comfort of patients, which has certain clinical application value.

Objective To explore the feasibility and accuracy of using indocyanine green fluorescence (ICGF) to identify the intersegmental plane after ligation of the target pulmonary vein during thoracoscopic segmentectomy. Methods From December 2022 to June 2023, the patients with pulmonary nodules undergoing video-assisted thoracoscopic anatomical segmentectomy with intersegmental plane displayed using ICGF after ligation of the target pulmonary vein by the same medical team in our hospital were collected. Preoperative three-dimensional reconstruction was used to identify the target segment where the pulmonary nodule was located and the anatomical structure of the arteries, veins, and bronchi in the target segment. The intersegmental plane was first determined by the inflation-deflation method after the target pulmonary vein was ligated during the operation. During the waiting period, the target artery and bronchus could be separated but not cut off. The inflation-deflation boundary was marked by electrocoagulation, and then ICGF was injected via peripheral vein to identify the intersegmental plane again, and the consistency of the two intersegmental planes was finally evaluated. Results Finally 32 patients were collected, including 14 males and 18 females, with an average age of 58.69±11.84 years, ranging from 25 to 76 years. The intersegmental plane determined by inflation-deflation method was basically consistent with ICGF method in all patients. All the 32 patients successfully completed uniportal thoracoscopic segmentectomy without ICGF-related complications or perioperative death. The average operation time was 98.59±20.72 min, the average intraoperative blood loss was 45.31±35.65 mL, and the average postoperative chest tube duration was 3.50±1.16 days. The average postoperative hospital stay was 4.66±1.29 days, and the average tumor margin width was 26.96±5.86 mm. Conclusion The ICGF can safely and accurately identify the intersegmental plane by target pulmonary venous preferential ligation in thoracoscopic segmentectomy, which is a useful exploration and important supplement to the simplified thoracoscopic anatomical segmentectomy.