Objective To study the relationship between X-linked inhibitor of apoptosis protein (XIAP) expression and transitional cell carcinoma(TCC) development. Methods Forty-three TCC tissues and 12 normal transitional epithelial tissues were applied to detect XIAP expression by semi-quantitative RT-PCR, immunohistochemistry and western blot. The data were statistically analyzed by using SPSS11.5 according to the 2 groups (TCC and normal transitional epithelial) as well as the dif-ferent subgroups (tumor stage, grade, single or multiple tumor, primary or recurrence tumor). Results XIAP expression in TCC tissues was higher than in normal transitional epithelial tissues(im-munohistochemistry: 22±5 and 16±2, Western blot:1.21±0. 15 and 0. 61±0.24, mRNA: 1.17± 0. 30 and 0. 75±0. 17, P<0. 05). In the bladder tumors group, XIAP expression in recurrence tumors was higher than in primary tumors(immunohistochemistry: 24±3 and 20±3, Western blot: 1.66±0.28 and 1.10±0. 23, mRNA: 1.44±0. 27 and 1.05±0. 23, P<0. 05). However, there were no significant differences according to the tumor stage and tumor grade as well as tumor multi-plicity or not. Conclusion XIAP expression might serve as a biomarker in TCC diagnosis and recur-rence prediction.

Objective To study the missed diagnosis of colorectal polyps during colonoscopy and its risk factors.Methods Data of 655 patients who underwent repeated co]onoscopy in 3 months (90 days) were analyzed in three endoscopy centers in Shenzhen.Miss rates of polyps and patients were calculated.Logistic regression analysis was used to identify the suspected risk factors associated with the miss rate including gender,age,symptoms of patient and number,shape,location of polyps.Results A total of 459 polyps(20.47％,459/2 242) in 224 patients(34.20％,204/655) were missed in overall 1 783 polyps within 655 patients.The patient miss rate increased with the polyp count increasing from 1 to 4,but with no significant differences.Polyp count of more than 5 was the independent risk factor for patient miss rate during colonoscopy(OR=4.98,P=0.00).Polyps in males were easier to be missed than those in females (OR =1.76,P =0.00).Size less than 5 mm was the independent risk factor for missed diagnosis during colonoscopy(OR=2.94,P=0.00).The flat type(Yamada Ⅰ,Ⅱ) was also the independent risk factor(OR=2.72,P=0.01;OR=3.23,P=0.00 respectively).Conclusion The miss rate of polyps is related to gender,basic polyp count,the size and shape of polyp.Male with multiple polyps and polyps with flat type and small size tend to be missed.

Objective To investigate the mechanisms of oridonin inducing apoptosis on acute leukeamia NB4 cells and its mechanism. Methods NB4 cells in culture medium in vitro were given with different concentrations (8, 16, 24, and 32 μmol/L) of oridonin. The inhibitory rate of the cells was measured by MTT assay, cell apoptotic rate was detected by flow cytometry (FCM), morphology of apoptosis was observed by Hoechst 33258 fluorescence staining, DNA fragmentation was assayed by agarose gel electrophoresis, caspase-3 expression was detected by Western blotting, and caspase-3 activity was assayed with colorimetric assay kit before and after apoptosis occurred. Results Oridonin (over 16 μmol/L) could inhibit the growth of NB4 cells and cause apoptosis significantly, the suppression was both in a timeand dose-dependent manner. Marked changes of apoptosis including condensation of chromatin and nuclear fragmentation were observed very clearly by Hoechst 33258 fluorescence staining and a characteristic "ladder" of DNA fragments was elicited by agarose gel electrophoresis; Western blot analysis revealed that caspase-3 was activated by the loss of caspase-3 proenzyme (32 kDa) and the appearance of its 20 kDa subunit, and that along with the apoptotic process caspase-3 activity was increased concurrently. Conclusion Oridonin can induce apoptosis in NB4 cells via activation of caspase-3. These results will provide laboratory evidence for the clinical treatment of acute leukemia with oridonin.

Objective To evaluate the long-term therapeutic results and the safety of nephronsparing surgery(NSS) for the treatment of renal cell carcinoma. Methods Clinical data of 243 NSSfor renal cell carcinoma were retrospectively analysed. Of them, 159 were males and 84 were femaleswith average age of 58 years (range from 24 ?77 years). The average tumor size was 3. 4 cm (rangefrom 1.1 to 6. 7 cm). Three cases were solitary renal cell carcinoma, 11 were bilateral renal cell carcinoma; 237 cases were in stage T_(1a). and 6 cases were in stage T_(1b). No lymph node and distant metastasis, no renal vein cancer tumor embolus and inferior vena cava tumor embolus was found. Postoperative follow-up was carried out by ultrasound, CT and renal function. Cancer specific survival was estimated using Kaplan-Meier method and log-rank test. Results After a mean 31 months (1-147months) follow-up, long-term follow-up data were obtained in 232 cases because the other 11 did notlive in Dalian, 52 were treated with interferon. Four of the 232 patients treated with NSS had died:1died from lung cancer 16 months after lung cancer treatment, the other 3 died from cardiovascular diseases. The total survival rate and cancer specific survival rate were 98. 3% and 100. 0%, respectively.Local tumor recurrences were detected in 5 patients and tumor metastasis was detected in 1 patient.The recurrence rate was 2. 2%, and the metastasis rate was 0. 4%. The complications included temporary renal failure and urine leakage. The complication rate was 5. 6%. Conclusions NSS for renalcell carcinoma is a safe and feasible treatment option. It has the advantages of low local recurrence,good long-term survival rate and low complication rate. NSS can maximally reserve functional nephron, reduce the risk of chronic renal failure, preserve patient's quality of life and increase patient'ssatisfaction.

To obtain bovine adipose-derived stem cells (ADSCs), bovine ADSCs were digested in collagenase type I solution. The growth curve of ADSCs was checked by cell counting. Chromosome analysis was checked. The molecular markers of ADSCs were detected with immunofluorescence staining. The morphology of ADSCs was identical to fibroblast like and the cells showed active proliferative ability. Vimentin, CD49d and CD13 antigens were detected, but CD34 antigen was negative. Alkaline phosphatase activity was greater in ADSCs during calcification, and Alizarin Red staining was positive. Lipid droplets were apparent around cells during adipogenesis, and Oil Red-O staining was positive. The results demonstrated that ADSCs could be used as seed cells for tissue engineering due to the simple isolation, differentiation and stable and active growth.
Adipose Tissue ; cytology ; Animals ; Cattle ; Cell Differentiation ; physiology ; Cell Separation ; Cells, Cultured ; Mesenchymal Stromal Cells ; cytology ; Tissue Engineering ; methods

OBJECTIVETo investigate the apoptosis inducing effects of ponicidin (PON) on leukemic K562 cells and its mechanisms of action.

METHODK562 cells in culture medium in vitro were given different concentrations of PON (10-50 micromol x L(-1)) for 24, 48 and 72 h. The inhibitory rate of the cells was measured by MTT assay, cell apoptotic rates were detected by flow cytometry (FCM) using Annexin V staining after K562 cells were treated with different concentrations of PON for 72 hours, and cell morphology was observed by Wright-Giemsa staining. Western blot was used to detect caspase-3 and poly(ADP-ribose) polymerase (PARP) expression, and the protein levels in mitogen-activated protein kinase signaling pathways (MAPKs, p-P38, p-ERK and p-JNK) as well as p-AKT and p-P85 in PI3K/AKT signaling pathways were also detected.

RESULTPON (over 30 micromol x L(-1)) could inhibit the growth of K562 cells in both time- and dose-dependent manner. FCM analysis revealed that apoptotic cells were gradually increased in a dose-dependent manner after treatment for 72 hours, and that marked morphological changes of cell apoptosis such as condensation of chromatin was clearly observed by Wright-Giemsa staining after treatment by 50 micromol x L(-1) PON. Western blot showed cleavage of the caspase-3 zymogen protein (32 kD), with the appearance of its 17 kD subunit, and a cleaved 89 kD fragment of 116 kD PARP was also found. Furthermore, Western blotting also showed that expression of p-AKT and p-P85 in PI3K/AKT signaling pathways was downregulated dramatically whereas the expression of p-P38 as well as p-ERK and p-JNK remained unchanged after the cells were treated by PON for 48 h.

CONCLUSIONThe results demonstrate that PON exhibits in vitro anti-leukemia effect by induction of apoptosis in K562 cells, and that PON induced apoptosis in K562 cells mainly related to activation of caspase-3 as well as inactivation of PI3K/AKT signaling pathway via down regulation of the expression of p-AKT and p-P85 protein levels. These results provide strong laboratory evidence for further anti-leukemia trials of PON.

Apoptosis ; drug effects ; Blotting, Western ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Diterpenes ; pharmacology ; Humans ; Poly(ADP-ribose) Polymerases ; metabolism ; Signal Transduction ; drug effects

Objective To evaluate the effect of esketamine for cesarean section under epidural an-esthesia and maternal and fetal outcome.Methods The parturients all were singleton pregnancies at term and scheduled for elective cesarean delivery.Ninety parturients undergoing elective cesarean delivery under epidural anesthesia,aged 20-40 years,BMI 18-35 kg/m2,ASA physical status Ⅱ,were randomly alloca-ted into two groups:esketamine group(group K)and control group(group C),45 cases in each group.When the targeted upper sensory block level was achieved,esketamine 0.15 mg/kg was administered intra-venously for 1 minute before skin incision in group K,the same volume of normal saline was intravenously injected in group C.Visceral traction reaction during the operation was recorded.Apgar score was assessed 1 minute and 5 minutes after birth.The pH value,partial pressure of carbon dioxide,oxygen partial pressure,and lactic acid value of the neonatal umbilical vein blood were recorded.Adverse events such as nausea,vomiting,hallucination,dizziness and nightmare were monitored in the two groups.Results Compared with group C,visceral traction reaction was significantly lower in group K(P＜0.05).The neonatal Apgar score,pH value,carbon dioxide partial pressure,oxygen partial pressure,and lactic acid value of the neo-natal umbilical vein blood did not differ between the two groups.There were no differences in the incidence of nausea,vomiting and dizziness between the two groups.Hallucinations and nightmares were not encoun-tered in either group.Conclusion Esketamine 0.15 mg/kg can effectively reduce visceral traction reaction during cesarean delivery under epidural anesthesia.There was no significant effect on neonatal Apgar score and blood gas analysis,with no noticeable maternal and neonatal adverse effect.

Objective:To study the genetic profile of neonatal hyperbilirubinemia with unknown etiology in Guangdong Province and the clinical significance of jaundice-related genetic screening.Methods:From July to September, 2021, neonates with hyperbilirubinemia of unknown etiology born in different hospitals in Guangdong Province were studied. 24 neonatal jaundice-related exons were sequenced using targeted capture and high-throughput sequencing technology. The pathogenic variants were analyzed.Results:A total of 331 cases, 139 (42.0%) cases showed positive screening results with five diseases, including 65 (19.6%) cases of Gilbert syndrome, 48 (14.5%) cases of glucose-6-phosphate dehydrogenase (G6PD) deficiency,18 (5.4%) cases of sodium taurocholate cotransporting polypeptide deficiency, 4 (1.2%) cases of Citrin deficiency and 4 (1.2%) cases of Dubin-Johnson syndrome. 149 (45.0%) cases carried one or more genetic variants and 43 (13.0%) cases showed no clinically significant variants. The 8 high-frequency mutation loci (carrier rate >1%) are UGT1A1 gene c.211G>A and c.1091C>T, G6PD gene c.1466G>T and c.1478G>A, SLC10A1 gene c.800C>T, SLC25A13 gene c.852_855del TATG, HBB gene c.126_129delCTTT and c.316-197C>T.Conclusions:Genetic factors are important for neonatal hyperbilirubinemia with unknown etiology in Guangdong. The common pathogenic genes are UGT1A1, G6PD, SLC10A1, and SLC25A13 and the population carries high-frequency mutation loci. Therefore, genetic screening in neonates with hyperbilirubinemia of unknown etiology has important clinical significance.

Objective:To investigate the normal range of exhaled nitric oxide (FeNO) in 6-18-year-old children in China, so as to provide a data base for the establishment of FeNO standards for Chinese children.Methods:A multi-center study was conducted on 5 949 children aged 6-18 (3 101 males and 2 848 females) in 16 pro-vinces of 7 administrative districts in China.According to the technical standard recommended by American Thoracic Society/European Respiratory Association, FeNO was measured, and the relationship of FeNO with the sex, age, height, weight, body mass index and region was discussed.Results:The geometric mean FeNO value of Chinese children aged 6-18 was 14.1 ppb, and its 95% confidence interval (skewness distribution) was 1.0-38.2 ppb.The geometric mean FeNO values of children aged 6-11 and 12-18 were 13.1 ppb and 15.7 ppb, respectively, and their 95% confidence intervals (skewness distribution) were 1.0-38.1 ppb and 2.0-38.2 ppb.For children at and under 11 years old, FeNO decreased with age, with a mean decline of 1 ppb per year.The multiple linear regression results suggested that there was a significant correlation between FeNO and age for children aged 6-11, and FeNO of children aged 12-18 was significantly correlated with the gender, height, and region(all P<0.01). Conclusions:FeNO values of Chinese children and adolescents in this study are higher than those obtained by the previous study conducted from 2010 to 2012.For children aged 12-18, 16 ppb is recommended as the clinical cut-off point.For children at or under 11 years old, the influence of age on FeNO should be considered, and the cut-off point of FeNO decreases by 1 ppb as the age is reduced by one year.

Objective: To investigate the effects of long-term hyperglycemia on renal pathology of type 1 diabetic mellitus(T1DM) cynomolgus monkeys by establishing streptozotocin(STZ)-induced T1DM. Methods: Eight 4-year-old male cynomolgus monkeys were randomly divided into control and model groups. Four cynomolgus monkeys were used in the control group, and four cynomolgus monkeys were injected with streptozotocin to create a T1DM model. Hematoxylin-eosin(HE) staining and periodic acid-Schiff(PAS) staining were used to observe renal pathological changes.The histological characteristics and changes were observed under transmission electron microscope. Morphometric measurements were used to analyze the glomerular area, the thickness of the glomerular basement membrane, the proportion of glomerular podocyte foot processes and the average area of foot processes. Results: In comparison to the control group, the model group showed pathological changes in the kidneys, including increased glomerular area(P<0.01), thickened basement membrane, capillary loop compression, disordered arrangement of endothelial cell fenestrations, proliferation of mesangial cells, mesangial expansion, accumulation of mesangial matrix glycoproteins, a decrease in the number of foot processes(P<0.001), a decrease in the average size of the foot processes(P<0.05), widened podocyte slit diaphragm, and an increase in the number of fused foot processes(P<0.001). Conclusion Cynomolgus monkeys with T1DM suffer from renal pathological changes due to long-term hyperglycemia.