Objective:To evaluate the safety and efficacy of Delorme procedure for adults with full-thickness rectal prolapse.Methods:Clinical data of 17 adult patients suffering from full-thickness rectal prolapse undergoing Delorme procedure from June 2014 to May 2018 in Hangzhou Third Hospital were retrospectively analyzed. Patient characteristics, operative data, postoperative complications, recurrence of rectal prolapse, continence state and constipation state were evaluated.Results:Eleven patients were female, 6 patients were male with a mean age of (68 ± 9) years. Operations were successfully performed in these 17 cases. The operation time was (88 ± 16) minutes. The estimated blood loss during operation was (23 ± 9) ml. The postoperative time of hospital stay was (8 ± 1) d. Two complications in two patients were observed. There was no treatment related death. One recurrent case was observed during (16 ± 2) months follow-up. The preoperative and postoperative mean constipation score of five patients with fecal constipation were (23 ± 2) and (11 ± 3) respectively ( t = 9.51, P<0.01). The mean fecal incontinence score of six patients with fecal incontinence, before and after Delorme procedure, were (14 ± 2) and (6 ± 2) respectively ( t = 9.09, P<0.01). Conclusions:The Delorme procedure for adults with full-thickness rectal prolapse is a safe and effective surgery with less complications and low recurrence rate. The Delorme procedure may be one of the preferred option of perineal approach for adults with full-thickness rectal prolapse, but the long-term outcome of Delormer procedure and its effect on postoperative anal function need to be further studied.

Objective:To investigate the correlation between serum cystatin C (CysC) and clinical and pathological features of IgA nephropathy.Methods:Four hundred and twenty-one cases of primary IgA nephropathy diagnosed by renal biopsy in Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine from January 2010 to January 2021 were retrospectively analyzed. According to the serum CysC level at the time of renal biopsy, the patients were divided into high serum CysC group and normal serum CysC group, and the clinical data and pathological indices of the patients were compared. Spearman correlation analysis was used to analyze the correlation between estimated glomerular filtration rate (eGFR) and serum CysC. The clinicopathological factors related to the serum CysC level were analyzed by multiple linear regression. The area under the receiver operator characteristic curve (AUC) was used to evaluate the ability of serum CysC level to predict related pathological injury.Results:The age, prevalence of hypertension, serum creatinine, urea and uric acid levels of high serum CysC group were significantly higher than those of normal serum CysC group, while the eGFR level was significantly lower than that of normal serum CysC group ( P<0.05). Spearman correlation analysis showed that serum CysC was negatively correlated with eGFR ( r=-0.744, P<0.001). In terms of pathological injury, the degree of renal tubular atrophy and renal interstitial fibrosis (T) and renal arteriole wall thickening (A) in high serum CysC group were more serious than those in normal serum CysC group ( P<0.05). Multiple linear regression analysis showed that the prevalence of hypertension, serum creatinine, urea, uric acid, T and A were correlated with serum CysC levels (standard regression coefficient β=0.048, 0.299, 0.260, 0.134, 0.195, 0.068, respectively, P<0.05). After adding serum CysC on the basis of clinical features, the prediction efficiency of renal tubular atrophy and renal interstitial fibrosis was higher (AUC were 0.829 [95% CI 0.787-0.870], 0.847 [95% CI 0.808-0.886], P<0.05). Conclusions:Patients with older age, hypertension, poor renal function and severe pathological damage are more likely to have elevated serum CysC levels. Serum CysC was related to the prevalence of hypertension, creatinine, urea, uric acid, T and A. Combined with serum CysC level can effectively improve the ability prediction of T.

Objective:To evaluate the retinal differentiation ability of human induced pluripotent stem cells (hiPSCs) from various somatic cell sources.Methods:The hiPSCs lines BC1- green fluorescent protein (GFP) and Gibco obtained by blood cell reprogramming and the hiPSCs line UE017 obtained by urine cell reprogramming were used to induce retinal differentiation.The morphogenesis and development of retina were recorded with an optical microscope, and the expression of specific molecular markers of various cell subclasses in the retina was detected by immunofluorescence, and the efficiency of retinal differentiation of different cell lines was analyzed and compared.Results:All three hiPSC lines derived from blood and urine cells were able to be induced into three-dimensional (3D) retinal organoids, including neuroretina and retinal pigment epithelial cells.Retinal organoids simulated the development process of retina in vivo and gradually differentiated into all cell subtypes of retina, including retinal ganglion cells, photoreceptor cells, amacrine cells, horizontal cells, bipolar cells, Müller cells, and even formed lamellar structures.However, in terms of the efficiency of acquiring retinal organoids, the hiPSCs derived from blood were more efficient than those derived from urine. Conclusions:hiPSCs from both blood and urine somatic cells can differentiate into 3D retinal organoids, including all subtypes of retinal cells.The differentiation efficiency among lines is different.

Objective:To establish a fluorescent reporter human induced pluripotent stem cell line (hiPSCs) for monitoring the expression of visual system homeobox 2 ( VSX2). Methods:VSX2_small guide RNA (sgRNA) was inserted into vector PX459 to construct knockout plasmid, and the P2A-eGFP knock-in donor plasmid was conducted at the same time.The two plasmids were transfected into BC1-hiPSCs.Single cell clones were generated after treatment of puromycin.Correct insertion was confirmed by PCR and Sanger sequencing.The isogenicity of the parental and the reporter hiPSCs was confirmed by STR analysis and karyotyping.Pluripotency capacity of the reporter hiPSCs was analysed by reverse trascription PCR and immunofluorescence.Three-germ-layer formation experiment was carried out to analyse the multi-lineage differentiation ability of the reporter hiPSCs.The reporter hiPSCs were further differentiated to obtain three-dimension (3D) retinal organoids, and immunofluorescence was used to identify the co-localization of the enhanced green fluorescent protein (eGFP) and VSX2.Results:A VSX2 eGFP reporter hiPSC clone was successfully obtained by CRISPR/Cas9 technology, which was consistent with the parental hiPSCs (BC1-hiPSCs) in morphology, without any chromosomal aberrations or cell line cross-contamination.Reverse transcription PCR assay and immunofluorescence showed obvious positive expressions of iPSCs markers in BC1- VSX2 eGFP-iPSCs, including NANOG, OCT4, SOX2, DNMT3B and GDF3 mRNA as well as NANOG, OCT4, SSEA4 and TRA-1-60 protein.The α-fetoprotein (AFP), α-smooth muscle actin (α-SMA) and neuronal class Ⅲ β-tubulin (TUJ1) were expressed in endoderm, mesoderm and ectoderm, respeetively, derived from BC1- VSX2 eGFP-iPSCs, and eGFP and VSX2 were co-stained in the neural retinal layer of 3D retinal organoids derived from BC1- VSX2 eGFP-iPSCs by immunofluorescence. Conclusions:VSX2 fluorescent reporter hiPSCs is successfully generated, which can monitor the temporal and spatial expression changes of VSX2 protein in real time, providing a powerful tool for evaluation of retina development mechanism and cell therapy.

Objective:To investigate the effects of total minimally invasive surgery (TMA) and mixed minimally invasive/open surgery (HMOA) on perioperative conditions and long-term efficacy of patients with rectal cancer.Methods:The clinical data of 240 patients with rectal cancer treated with minimally invasive surgery from January 2014 to August 2018 in Hangzhou Third People′s Hospital were retrospectively analyzed. Among them, 110 patients were treated with TMA (TMA group) and 130 patients were treated with HMOA (HMOA group). The relevant indexes of patients before and after surgery were collected and analyzed.Results:The operation time in TMA group was significantly longer than that in HMOA group: (312.5 ± 20.3) min vs. (210.8 ± 15.2) min, the length of hospital stay was significantly shorter than that in HMOA group: (4.0 ± 0.5) d vs. (6.8 ± 1.0) d, and there were statistical differences ( P<0.01); there were no statistical differences in low anterior resection and surgical procedures, ileostomy, open surgery, postoperative complications, reoperation, morphine dosage at 3 d after surgery and readmission between 2 groups ( P>0.05). Multivariate Cox analysis result showed that BMI ≥ 30 kg/m 2 ( OR=4.11, 95% CI 1.68 to 9.72, P<0.01), TMA ( OR=0.13, 95% CI 0.06 to 0.42, P<0.01), delayed bowel obstruction ( OR=13.6, 95% CI 1.59 to 110.56, P<0.05) and reoperation ( OR=15.32, 95% CI 5.52 to 42.56, P<0.01) were independent risk factors of prolonged hospital stay in patients with rectal cancer. The patients were followed up for 15 to 42 (29.5 ± 0.2) months, and there were no statistical differences in 3-year overall survival (OS) rate and 3-year disease-free survival (DFS) rate between HMOA group and TMA group (92.5% vs. 92.8% and 79.6% vs. 85.5%, HR=1.20 and 0.75, 95% CI 0.35 to 3.14 and 0.28 to 1.34, P=0.98 and 0.25). Conclusions:Patients with rectal cancer treated with TMA have the advantages of shorter hospital stay and shorter short-term effects compared with those treated with HMOA. However, the long-term effects of the two minimally invasive procedures are similar.

Objective:To study the anti-tumor efficacy of endostar microbubble combined with focused ultrasound radiation in colon cancer liver metastases.Method:29 mice with colon cancer liver metastasis were randomly divided into four groups. Group 1(8 mice), as the control group. Group 2(7 mice) were treated only with ultrasonic radiation. Group 3 (7 mice) treated with the ultrasonic radiation combined with SonoVue microbubbles without carrying any medicine. Group 4(7 mice), treated with the ultrasonic radiation combined with microbubbles carrying endostar. The mice were sacrificed and the tumor specimens were weighted on the 12 days after ultrasound radiation. Immunohistochemistry was used to assess CD34 expression within the metastatic tumor.Results:The tumor weight in group 4 (0.79±0.49)g was significantly lower than that in group 1 (2.67±0.61)g, group 2 (2.60±0.60)g and group 3 (1.74±0.33)g ( F=20.629, P<0.01). The liver metastatic tumor weight in group 4(0.55±0.16) g was much lower than that in group 1 (1.47±0.22)g, group 2(1.42±0.28) g and group 3 (0.95±0.27)g ( F=23.758, P<0.01). There was no obvious difference among the four groups in the number of nodules of metastatic tumor in liver ( F=0.167, P=0.918). The level of CD34 in group 4 were (8 037±1 708) , significantly lower than that in any other group, ( F=15.779, P<0.01). Conclusion:Endostar microbubble combined with focused ultrasound radiation decreases tumor angiogenesis in liver metastasis, and inhibits the growth of both primary and metastatic tumor.

OBJECTIVETo explore the impact of neutrophil gelatinase-associated lipocalin (NGAL) knockdown by NGAL siRNA encapsulated with urocanic acid-modified chitosan nanoparticles (UAC) on the proliferation, migration and apoptosis of human colon cancer cells.

METHODSNGAL siRNA was encapsulated by UAC and chitosan (CTS) respectively, and then was transfected into human colon cancer cell lines HT29. The NGAL mRNA was detected by real-time quantitative PCR (RT-QPCR). Relationships of NGAL gene silencing with the proliferation, migration and apoptosis of HT29 cell were analyzed.

RESULTSUnder the fluorescence microscope, the transfection efficiency of siRNA in UAC group was (37.52±7.17)%, which was significantly higher than (11.32±3.39)% in CTS group (t=6.102, P=0.005). Forty-eight hours after transfection, RT-QPCR examination showed that the level of NGAL mRNA expression was 0.350 in UAC group and 0.529 in CTS group with significant difference (t=-3.743, P=0.02), meanwhile both levels were significantly lower as compared to control group(F=163.538, P<0.001). Proliferation analysis revealed that after silencing NGAL gene, proliferation rate of UAC group and CTS group was slightly lower than control group, and no significant differences were found (F=9.520, P=0.438). However, migration assay demonstrated that the 24-hour migration rate of UAC group and CTS group was significantly lower than that of control group (F=6.756, P=0.029), meanwhile the migration rate of UAC group was slightly lower than that of CTS group [(77.90±7.14)% vs. (87.67±3.98)%, t=-1.704, P=0.164]. Apoptosis detection revealed that the apoptosis rate in UAC group was significantly higher than that in CTS group and the control group 2 days after transfection [(15.800±1.054)% vs. (12.900±0.656)%, (11.933±1.914)%, F=7.004, P=0.027].

CONCLUSIONSThe encapsulated ability and transfection efficiency of chitosan modified by urocanic acid elevate significantly. Silencing NGAL gene by UAC carrier can down-regulate the expression of NGAL mRNA in HT29 colon cell line, inhibit their migration and facilitate their apoptosis.

Objective To investigate the prevalence of metabolic syndrome (MS) among adults in rural areas of Ningxia Hui autonomous region.Methods A cross-sectional study was conducted in 10 639 adults enrolled with a multistage method from Jingyuan County.The MS was identified according to Chinese type 2 diabetes prevention guide (2013).Results Among all the subjects, 17.4% of them met the MS definition with the standardized prevalence of 14.7% after adjustment of sex and age.The prevalence and standardized rate of MS in men were 19.9% and 17.3%, and in women were 15.3% and 13.5%.The prevalence of MS in men was higher than that in women(P<0.001) and increased with aging in both genders.The prevalence and standardized rate of abdominal obesity,hyperglycemia,hypertension,high triglycerides,and low HDL-C were 19.5% and 16.7%, 15.0% and 12.9%, 42.0% and 37.1%, 25.8% and 23.1%, 28.5% and 27.7%,respectively.The rate of abdominal obesity was higher in women than in men (20.5% vs 18.2%, P=0.004), whereas the rate of hypertension, high triglycerides, and low HDL-C were higher in men than in women (all P<0.01).The prevalence of having one parameter of the MS was 68.4%.Conclusion The prevalence of MS is higher in rural areas of Ningxia Hui autonomous region, suggesting that a series of comprehensive prevention measures should be carried out to prevent and control the MS so as to improve the public health conditions in rural areas.

Objective To explore the effect of NGAL knockdown by NGAL siRNA encapsulated with urocanic acid-modified chitosan nanoparticles (UAC).MethodsNGAL siRNA encapsulated by UAC and chitosan (CTS) respectively, which were then used to transfect human colon cancer cell lines HT29.The expression level of NGAL protein were detected by Enzyme Linked Immunosorbent Assay(ELISA).ResultsThe ELISA study revealed that the expression level of NGAL protein in UAC group(average 0.583μg/L) was significantly lower than in CTS group (average 0.772μg/L) and control group(average 1.071μg/L) (P<0.05).ConclusionThe NGAL expression of mRNA and protein in HT29 cells could be down-regulated by siRNA encapsulated by UAC.