Objective To investigate the effect of dysadherin gene silencing on metastasis and invasion in pancreatic cancer cell line PANC1,BxPC3 in vitro.Methods PANC1 and BxPC3 cells were divided into dysa group,negative siRNA control group(HK),liposomes control group(control),dysa group and HK group were tranfected by dysadherin siRNA and Negative siRNA,respectively.The expression of dysadherin mRNA and protein were detected by RT-PCR and immunohistochemical method.Transwell test was used to evaluate the invasion ability of pancreatic cancer cells.Results After transfected by dysadherin siRNA,the dysadherin mRNA levels in PANC1 and Bxpc3 cells were decreased by 95.4% and 52.1%.The expression of dysadherin protein was also down-regulated by 91.2% and 83.6%,respectively,when compared with HK groups (P＜0.05 ).The numbers of invasive cells migrated in Transwell in PANC1 cells control group,HK group and dysa group were 163.2±15.5,154.4±17.3 and 53.6±7.9;the numbers of invasive cells in BxPC cells control group,HK group and dysa group were 30.7±3.2,27.5±2.8 and 4.7±2.4,respectively.The numbers in dysa group were significantly lower than those of HK group and control group (P＜0.01 ).Conclusions Silencing the dysadherin gene of PANC1,BxPC3 by RNA interference could significantly inhibit the invasive and migratory ability of canceroas cells.

ObjectiveTo investigate the biological effects and its mechanisms of ascorbic acid on pancreatic cancer PANC1 cells. Methods PANC1 cells were treated by ascorbic acid of different concentrations (0 ～40 mmol/L) for 24,48,72 hours.The proliferation of PANC1 cells was analyzed by MTT method; cell cycle and apoptosis were assessed by flow cytometry (FCM); inverted microscopy and transmission electron microscopy were used to observe cell morphology. The membrane potential of mitochondria were mearured by with JC-1 staining and FCM.Meanwhile,the changes of cell morphology and mitochondrial membrane potential induced by ascorbic acid after pretreatment with hydrogen peroxidescavenging enzyme (catalase) and red blood cells were also detected. Results Ascorbic acid in pharmacologic concentrations selectively inhibited the proliferation of PANC1 cells in a dose and time dependent manner.PANC1 cells were arrested in G2/M phase after treatment with 5 mmol/L ascorbic acid [ (32.55 ± 7.14)％ vs (22.00 ±1.27)％,t =5.808,P＜0.05],but there was no changes on apoptosis rate [ (1.98 ± 1.80)％ vs (1.09 ±0.16)％ ].Inverted microscope and transmission electron microscopy showed that oncosislike cell death of PANC1 cells was induced after treatment with ≥5 mmol/L ascorbic acid.Mitochondrial membrane potential of PANC1 cells was significantly lower than that of the control group in a dose dependent manner.The descent of mitochondrial membrane potential was significantly inhibited by pretreatment with catalase and red blood cells,and the degree of cell oncosis was attenuated.ConclusionsAscorbic acid significantly inhibited the proliferation of pancreatic cancer PANC1 cells in vitro.Ascorbic acid induced PANC1 cell oncosis,but not apoptosis.The possible mechanisms of inducing oncosis may be related to the descent of mitochondrial membrane potential.

Objective To explore the value of urine formaldehyde test in the diagnosis of Alzheimer's disease (AD),and the influential factors of urine formaldehyde level in AD patients.Methods A total of 52 AD patients and 53 cognitively normal controls were recruited in a cohort study.All subjects were no less than 65 years old,and those with acute infection,or dysfunction in heart,liver or kidneys were excluded.The impact of age,gender,onset age,MMSE score,NPI score,MTA score,and ApoE ε4 gene on urine formaldehyde of AD patients were analyzed by multiple regression analysis.Results Urine formaldehyde level of AD group was statistically higher than that of cognitively normal control group ((13.27±4.16)μmol/L vs (10.76±4.47)μmol/L,t=2.99,P=0.15).Urine formaldehyde of AD patients was statistically negatively correlated with MMSE score (β=-0.35,P=0.03) and MTA score (β=-0.38,P=0.02).The impact of onset age,neuropsychiatric disorders and ApoE ε4 gene on urine formaldehyde of AD patients was not statistically significant(all P>0.05).Conclusion Urine formaldehyde level is worthwhile to be explored as a marker in AD diagnosis and severity assessment.

Objective To explore the expression characteristic of serum mir-16,mir-21,mir-29a, mir-92a and mir-143 in colorectal cancer , evaluate the clinical significance of candidate miRNAs in CRC diagnosis.Methods Case-control study was used.The expression levels of serum mir-16,mir-21,mir-29a, mir-92a and mir-143 from 50 CRC patients and 27 normal controls were detected by real-time PCR, the levels of miRNAs expression in serum of 8 CRC patients 1 day before and 7 days after radical surgery were analyzed.The sensitivity and specificity of serum miRNAs expression for the diagnosis of colorectal cancer were evaluated using the receiver operating characteristic ( ROC) curve.Results The expression level of mir-16, miR-21, mir-92a in serum of patients with CRC were (75.55 ±37.73), (35.96 ±23.81), (24.79 ±8.97) fmol/ml, significantly higher than that of the healthy control group (32.73 ±18.94), (24.36 ±13.27), (16.36 ±5.58) fmol/ml (tmir-16 =2.77, tmir-21 =2.34, tmir-92a =3.85,P <0.05).However, the expression levels of mir-29a and mir-143 showed no significant difference between two groups (tmir-29a=-0.17, tmir-143 =1.59,P>0.05).In addition, serum miR-16 and miR-92a levels of CRC patients 7 days after tumor resection were (36.02 ±19.95), (14.82 ±7.78) fmol/ml, significantly lower than that before operation (62.18 ±34.17), (24.06 ±12.99) fmol/ml (tmir-16 =3.59, tmir-92a =2.60,P<0.05).Area under the ROC curve ( AUC) of combined detection of mir-16, miR-21and mir-92a was 0.877, the sensitivity and specificity were 88% and 85%, respectively, which was higher than any of 3 miRNAs alone and the conventional tumor marker CEA.Conclusion Combination of mir-16, mir-21 and mir-92a in the diagnosis of CRC shows a better sensitivity and specificity , which would be expected as new tumor biomarkers for noninvasive diagnosis and monitoring progression of colorectal cancer.

BACKGROUND:After internal fixation, limb fracture nonunion is the most common complication. Many factors affect fracture healing, but in recent years researchers have found that serum leptin may be involved in the process of fracture healing to regulate a variety of metabolisms. OBJECTIVE:To analyze the relationship between the changes of serum leptin and fracture healing in patients with long tubular fracture of the limbs after internal fixation. METHODS:Sixty patients with long tubular bone fracture who underwent internal fixation treatment were selected, and divided into two groups, union group (n=30) and nonunion group (n=30), according to the degree of fracture healing at 8 months after operation. Another 30 healthy volunteers served as normal control group. Peripheral blood samples were extracted before and after internal fixation to detect the changes in serum leptin levels using ELISA. RESULTS AND CONCLUSION:Preoperative serum leptin level was higher in the union group and nonunion group than the normal control group as wel as higher in the union group than the nonunion group (P < 0.05). There was no difference in the serum leptin levels in the union group before and after operation, but the nonunion group had a higher preoperative serum leptin level than the postoperative level. These findings indicate that the serum leptin may have an influence on fracture healing.

An early embryo co-culture system with human decidual stromal cells was established to study its effect on early embryonic cleavage and growth in vitro. Three hundred and eight 2-cell mouse embryos were co-cultured with human decidual stromal cell monolayer in MEM+0.4%bovine serum albumin (BSA) and 163 embryos cultured in MEM+15 % FCS alone as control. Among the mouse 2-cell embryos co-cultured with human decidual stromal cells, 72.73% developed to the morula stage and 67.21% cavitated to blastocysts with 59.74 % hatching, as compared with 61.34% to morula stage, 48.47% to blastocysts and none hatching in the controls,respectively. Co-cultured embryos cleaved slightly faster than controls and showed no or less fragmentation than those in the control. These results suggested that human decidual stromal cells can support early embryonic development and yield a reasonable number of embryos with good quality up to blastocyst stage.

An early embryo co-culture system with human decidual stromal cells was established to study its effect on early embryonic cleavage and growth in vitro. Three hundred and eight 2-cell mouse embryos were co-cultured with human decidual stromal cell monolayer in MEM+0.4%bovine serum albumin (BSA) and 163 embryos cultured in MEM+15 % FCS alone as control. Among the mouse 2-cell embryos co-cultured with human decidual stromal cells, 72.73% developed to the morula stage and 67.21% cavitated to blastocysts with 59.74 % hatching, as compared with 61.34% to morula stage, 48.47% to blastocysts and none hatching in the controls,respectively. Co-cultured embryos cleaved slightly faster than controls and showed no or less fragmentation than those in the control. These results suggested that human decidual stromal cells can support early embryonic development and yield a reasonable number of embryos with good quality up to blastocyst stage.

The superiority of the cumulative outcomes of day 5/6 embryo transfer to those of day 2/3 embryo transfer in infertile couples has been debated.This retrospective study included data collected from 1051 patients from July 2011 to June 2014.Multiple maternal baseline covariates were subjected to propensity score matching analysis,and each day 5/6 group woman was matched to one day 2/3 group woman.A systematic meta-analysis was conducted to validate the results.After matching was completed,217 patients on the day 2/3 group were matched with those on the day 5/6 group,and no significant differences in the baseline characteristics were observed between the two groups.The cumulative pregnancy rate (57.14％ vs.53.46％,OR 1.16,95％ CI 0.79-1.70) and cumulative live birth rate (53.00％ vs.49.77％,OR 1.14,95％ CI 0.78-1.66) of day 5/6 embryo transfers were higher than those of day 2/3 embryo transfers,but this difference was not significant.The mean cycles per live birth and mean days per live birth in the day 5/6 group were significantly lower than those in the day 2/3 group.This study demonstrated that day 5/6 embryo transfer is a more cost-effective and time-efficient policy than day 2/3 embryo transfer to produce a live baby.

Objective:To investigate the therapeutic effect of exosomes derived from remote ischemic conditioning on neurological dysfunction after cardiopulmonary resuscitation in a rat model of cardiac arrest and the relationship with glycocalyx protection.Methods:Exosomes were isolated from the blood of healthy adult male Sprague-Dawley rats using ultracentrifugation after undergoing remote ischemic conditioning for use as intervention drugs. Nanoparticle tracking analysis technology was used for exosome detection. Thirty-six adult male Sprague-Dawley rats were randomly assigned to 3 groups ( n=12 each) :Sham group, Control group and Exosome group. Cardiac arrest was induced by asphyxia for 7 min in the Control and Exosome groups. Placebo or exosomes (1×10 10 Particles) were infused intravenously at 5 min after the rats had returned of spontaneous circulation. Neuropsychological deficit score (NDS), open field test, Y maze and Morris water maze were used to assess neurological outcomes. The levels of plasma Hyaluronic acid (HA) and syndecan-1 (Sdc-1) were detected by Elisa. The expression levels of matrix metalloproteinase-2/9 (MMP-2/9) in hippocampal CA1 region were detected by Western blot. Results:After undergoing remote ischemic conditioning, the plasma levels of exosomes were elevated in rats compared to normal rats. Compared with the control group, the behavioral experiment of rats in the exosomes group were significantly improved, as evidenced by an increase in horizontal locomotor distance (5.86±2.89 vs. 17.53±5.51, P< 0.05), an increase in the correct rate of spontaneous alternation (13.29±15.07 vs. 42.63±10.25, P< 0.05), and a shortening of avoidance latency (25.83±8.54 vs. 13.49±4.55, P< 0.05). Plasma HA and Sdc-1 levels were significantly lower 24 h after resuscitation (HA: 26.34±9.83 vs. 14.84±6.26, P< 0.05; Sdc-1: 0.05±0.03 vs. 0.02±0.02, P<0.05), along with significantly lower MMP-2/9 levels in hippocampal tissue. Conclusions:Exosomes extracted from the plasma of rats undergoing remote ischemic conditioning can improve neurological dysfunction after cardiac arrest in rats, and the mechanism may be related to the inhibition of metalloproteinases and the reduction of endothelial glycocalyx degradation.

Objective:To explore the effects of the first dorsal metatarsal artery terminal branch flaps in repairing skin and soft tissue defects of fingers.Methods:The study was a retrospective observational study. From October 2021 to December 2022, 44 patients with skin and soft tissue defects in 55 fingers who met the inclusion criteria were admitted to Suzhou Ruihua Orthopedic Hospital. There were 39 males (48 fingers) and 5 females (7 fingers), aged 18 to 54 years. The single wound area after debridement ranged from 1.5 cm×1.0 cm to 3.0 cm×2.0 cm. The color Doppler ultrasonography was performed before operation to locate the first dorsal metatarsal artery and its terminal branches, and a first dorsal metatarsal artery terminal branch flap was designed according to the wound condition, with the area of harvested single flap ranged from 1.7 cm×1.2 cm to 3.2 cm×2.2 cm. The wounds in the flap donor areas were transplanted with full-thickness skin grafts from ipsilateral inner calf. The type of flap was recorded, and the diameter of the terminal branch of the first dorsal metatarsal artery was measured during operation. The survival of the flap was observed one week after operation. The wound healing in the flap donor and recipient areas was observed two weeks after operation. At the last follow-up, the functional recovery of the affected fingers was evaluated by the trial standards for evaluation of partial function of upper extremity by the Hand Surgery Society of Chinese Medical Association, the sensory function of the flap was evaluated using the sensory function evaluation standard of British Medical Research Council, the scar in the donor and recipient areas of the flap was evaluated using the Vancouver scar scale (VSS), and the Allen test was conducted in the toe of flap donor area to evaluate the blood flow.Results:The monoblock type flaps in 31 patients and flow-through type flaps in 2 patients were used to repair wounds in single finger, 2 monoblock type flaps in 8 patients were used to repair wounds in 2 fingers at the same time, and the single-pedicle and two-flap type flaps in 3 patients were used to repair wounds in 2 fingers at the same time. The diameter of the fibular terminal branch of the first dorsal metatarsal artery ranged from 0.40 to 1.10 mm, and the diameter of the tibial terminal branch of the first dorsal metatarsal artery ranged from 0.70 to 0.75 mm. All the flaps survived at one week after operation, and all the wounds demonstrated optimal healing in the flap donor and recipient areas at two weeks after operation. All patients were followed up for 6 to 18 months. At the last follow-up, the functional recovery of 48 fingers was evaluated as excellent, and the functional recovery of 7 fingers was evaluated as good; the sensory function of 8 flaps was rated as S2, and the sensory function of 47 flaps was rated as S3, and the two-point discrimination distance of the flaps was 8-14 mm; the VSS scores in the flap recipient areas ranged from 3 to 6, and the VSS scores in the flap donor areas ranged from 4 to 7; the Allen test result of the toes in the donor areas were all negative with normal blood flow.Conclusions:The first dorsal metatarsal artery terminal branch flaps have several advantages, including relatively hidden donor area, shallow anatomical level, simple intraoperative operation, and flexible flap design. The flap is incised without damaging the main artery of the toe, which can repair skin and soft tissue defects of the fingers and ensure the utmost protection of the toes in donor areas. The fingers exhibit improved appearance, texture, sensation, and function after operation.