Objective: To prepare the monoclonal antibody against ankyrin repeat domain 22（ANKRD22）and to investigate its expression in colorectal cancer tissues. Methods : The recombinant human ANKRD22 was expressed through E. coli and pET-42a and then used to immunize Balb/c mice after purification. Anti-human ANKRD22 specific monoclonal antibodies were selected by Western blotting with 293T cell lysate highly expressing ANKRD22 as antigen. The expression of ANKRD22 in the tissue microarrays of 112 patients with colorectal cancer was detected by immunohistochemical staining. Results : Four specific monoclonal antibodies against human ANKRD22 were screened out of 93 hybridoma cells,which reacted well with natural human ANKRD22. ANKRD22 was mainly distributed in the cytoplasm of colorectal cancer cells. In 112 cases of colorectal cancer,94 cases were detected positive for ANKRD22 expression,with the positive rate of 83.93%. The expression of ANKRD22 was statistically correlated with the expression of p53 and β-catenin（P<0.05）,but not with age,sex,location of tumors,AJCC stage,Dukes stage,degree of differentiation,lymph node metastasis and mismatch repair gene expression（P>0.05）. Conclusion The expression level of ANKRD22 was high in colorectal cancer. ANKRD22 might be involved in the carcinogenesis of colorectal epithelium and be a potential diagnostic marker.

Objective:To study the correlation between systemic immune inflammation index (SII) and prognosis of patients with hilar cholangiocarcinoma after surgical treatment.Methods:The clinical data of 181 patients with hilar cholangiocarcinoma treated by surgery at the First Affiliated Hospital of Zhengzhou University from January 2012 to December 2016 were retrospectively analyzed. There were 119 men and 62 women, with an average age of 62.4 years. SII was calculated using preoperative routine blood tests. Receiver operating characteristic (ROC) curve was used to obtain the optimal cutoff value of SII. The Kaplan-Meier method was used to draw survival curves and survival rates were compared by log-rank test. The Cox proportional risk model was used to analyze single and multiple factors.Results:The SII area under the ROC curve in predicting postoperative survival was 0.749(95% CI: 0.641-0.858), the optimal threshold was 412.6. Using this threshold, patients were divided into the low SII group (SII≤412.6, n=80) and the high SII group (SII>412.6, n=101). The 1, 3, and 5-year cumulative survival rates of patients in the low SII group were 87.5%, 57.5%, and 26.3%, which were significantly better than those of the high SII group of 71.3%, 39.6%, and 9.9% respectively ( P<0.05). Multivariate analysis showed that SII>412.6 ( HR=2.887, 95% CI: 2.256-7.903, P<0.05) was an independent risk factor for overall survival of patients with hilar cholangiocarcinoma. Conclusion:Preoperative SII had predictive values for postoperative survival of patients with hilar cholangiocarcinoma, SII>412.6 was an independent risk factor for postoperative survival.

Objective The purpose of this study is to observe the changes of immune cell subsets in lupus mice after umbilical cord mesenchymal stem cells (UC-MSCs) transplantation. Methods B6.MRL-Faslpr lupus mice were randomly divided into the following three groups: the UC-MSCs treated group, the fibroblast like synoviocytes (FLS) treated group and the untreated group. MSC (1×106) or FLS (1×106) were injected into the tail vein of lupus mice respectively. Four weeks after treatment, the spleen index was calculated. The pathological changes of kidney were assessed by HE staining. The frequencies of immune cell subsets in spleen and macrophage in kidney as well as abdominal cavity were analyzed by flow cytometry. Data were analyzed with t test. Results The spleen index of UC-MSCs treated lupus mice [(79 ±9) mg/10 g] and IgG level [(7.5±1.5) mg/ml] were significantly decreased when compared with FLS treated group [(147±23) mg/10 g, t=2.78, P<0.01] [(17.0 ±2.8) mg/ml, t=3.00, P<0.01] and the untreated group [(156 ±16) mg/10 g, t=4.29, P<0.01] [(16.7 ±1.6) mg/ml,t=4.01, P<0.01]. HE staining also showed that the pathological changes of kidney were alleviated after MSCs transplantation. In addition, the frequency of plasma cells in the untreated group [(2.61 2± 0.318)% vs (0.306±0.017)%, t=7.22, P<0.01] and the FLS treated group [(2.412±0.297)% vs (0.306±0.017)%, t=7.07, P<0.01] were markedly higher than MSCs treatment [(0.306 ±0.017)%]. Moreover, the frequency of CD25+Foxp3+/CD4+Treg in the MSCs treated group [(15.08±0.81)%] was significantly increased compared with the untreated group [(8.02 ±0.47)%, t=7.45, P<0.01] and FLS treated group [(8.80 ±0.23)%, t=7.39, P<0.01]. MSCs treatment resulted in a decrease in CXCR5+PD1+/CD4+Tfh and IFNγ+/CD4+Th1 subset, compared with the untreated group [(14.3±1.5)%vs (31.5±3.3)%, t=5.25, P<0.01] [(1.78±0.27)% vs (5.93±1.56)%, t=2.60, P<0.05] and the FLS treated group [(14.3±1.5)%vs (28.8±2.2)%, t=5.49, P<0.01] [(1.78±0.27)%vs (4.88±0.81)%, t=3.61, P<0.01]. The frequency of macrophage in kidney of the MSCs treated group [(3.52 ±0.37)%] was markedly increased compared with the untreated group[(1.58±0.29)%, t=3.25, P<0.01], while neither the IL4+/CD4+Th2 subset nor the IL17+/CD4+Th17subset and the frequency of macrophage in abdominal cavity showed significant changes in the three groups. Conclusion These findings suggest that the therapeutic effects of MSCs on lupus mice may mediate through increasing the frequency of spleen Treg and renal macrophage and decreasing the frequency of Tfh, Th1 and plasma cells.

OBJECTIVE: To explore the genetic etiology of three pedigrees with a gestational history of fetal renal anomalies. METHODS: Peripheral venous blood or skin samples were derived from the probands of the three pedigrees. Copy number variation sequencing (CNV-seq) was applied to detect alterations of genome CNVs. RESULTS: The patient from pedigree 1 and the fetuses from pedigrees 2 and 3 all carried a heterozygous 17q12 deletion, with the size ranging from 1.4 Mb to 1.48 Mb encompassing the HNF1B gene. CONCLUSION The diagnosis of 17q12 microdeletion may be difficult during fetal period for its variable phenotypes. Alterations of chromosomal copy numbers need to be excluded in such patients.
Chromosome Deletion ; Chromosomes, Human, Pair 17 ; genetics ; DNA Copy Number Variations ; Fetus ; Genetic Testing ; Hepatocyte Nuclear Factor 1-beta ; genetics ; Humans ; Pedigree ; Phenotype

Objective: This article provides an overview of our experience using indocyanine green angiography (ICGA) in breast reconstruction with abdominal flap to ascertain the application value of ICGA and its usage in decreasing postoperative complications. Methods: A total of 21 breast reconstructions with intraoperative ICGA were analyzed retrospectively, including 7 bilateral deep inferior epigastric perforator (DIEP) flaps, 5 pedicled transverse rectus abdominis myocutaneous (TRAM) flaps with contralateral free TRAM flaps, 4 pedicled TRAM flaps with contralateral DIEP flaps, 3 unilateral DIEP flaps and 2 unilateral pedicled TRAM flaps. According to different breast reconstruction methods, ICGA were applied respectively after flap harvesting and vessel anastomosis, in order to evaluate the blood supply of flaps and vessel perfusion. Results: A total of 52 ICGA were performed and recorded intraoperatively without any indocyanine green-associated complications. The operation methods were modified according to ICGA findings in 6 of 21 cases. The distal part of flaps were discarded due to poor perfusion in 2 cases (1 DIEP flap and 1 TRAM flap), additional free vessel anastomosis were needed in 2 cases to ensure sufficient blood supply, 2 vascular complication including 1 vascular occlusion and 1 vascular thrombosis were found and managed in time. During the follow-up (range from 3 to 30 months, median of 16 months), no vascular crisis was reported. All flaps survived satisfactorily without partial or whole flap necrosis or wound infection. Conclusions Intraoperative ICGA can provide real-time information of flap′s blood supply and vessel perfusion to evaluate the conditions of flaps and vascular anastomosis, which can help surgeons take actions accordingly to increase the successful rate of breast reconstruction.

Objective: To figure out the clinical application value of indocyanine green (ICG) lymphangiography in supermicrosurgical lymphaticovenular anastomosis. Methods: A total of 6 supermicrosurgical lymphaticovenular anastomosis with intraoperative ICG lymphangiography were performed during April 2015 to May 2017 and were analyzed retrospectively. All the cases are female (range from 30 to 54 years old, median of 46.5 years old), including 3 cases for prevention and 3 cases for treatment of lymphedema. Results: A total 6 supermicrosurgical lymphaticovenular anastomosis were performed with intraoperative ICG lymphangiography to make sure the influx of lymph fluid to the vein. During the median of 23 months follow-up (range from 7 to 32 months), the 3 preventive cases did not show upper limb lymphedema and the 3 theraputic cases were relieved at different levels. Conclusions Intraoperative ICG lymphangiography can provide real-time information to locate suitable lymph vessels and ascertain the anastomotic patency in supermicrosurgical lymphaticovenular anastomosis, thus improve the operation effectiveness.

Microsurgery techniques have allowed the development of many new therapeutic methods in plastic surgery, but are difficult to master without hard training. It is very important to set up a standardized microsurgery curriculum and training system for broadening surgical skills training and investigating the plastic surgery specialist training strategy. In our experiences, a series of training models are needed, like non-animal models, non- living animal models, live animal models and so on. This paper shows the training strategy for the primary stage of microsurgery training, non-animal model and non-living animal model training.

Microsurgery techniques have allowed the development of many new therapeutic methods in plastic surgery, but are difficult to master without hard training. It is very important to set up a standardized microsurgery curriculum and training system for broadening surgical skills training and investigating the plastic surgery specialist training strategy. In our experiences, a series of training models are needed, like non-animal models, non- living animal models, live animal models and so on. This paper shows the training strategy for the primary stage of microsurgery training, non-animal model and non-living animal model training.

Objective: The phenotype and genetics of three patients with autosomal recessive polycystic kidney disease (ARPKD) at childhood, teenage and advanced age were analyzed. Methods: Next generation sequencing (NGS) was applied to all the probands. PCR and Sanger sequencing were used to verify the suspicious gene variants screened by NGS in the probands and their family members, and one of the family got prenatal diagnosis. Results: Through NGS, PCR and Sanger sequencing, the 5-yr proband in pedigree 1 was shown to carry compound heterozygous variants of c. 5935G>A(p.G1979R) and c. 5428G>T(p.E1810X) of PKHD1, originated from his parents; In pedigree 2, the 17-ys proband was detected with c. 5512T>C(p.Y1838H) and c. 5935G>A(p.G1979R) variants of PKHD1 orginated from her parents, and her mother also got prenatal diagnosis during the second trimester; In pedigree 3, the 70-ys female proband was found with variants c. 11314C>T (p.R3772X) and c. 3860T>G (p.V1287G) of PKHD1. Conclusion The three pedigrees were diagnosed as ARPKD caused by PKHD1 variants. Five types of variants were detected, c. 5935G>A and c. 11314C>T were the known pathogenic variants, while c. 5512T>C, c. 5428G>T and c. 3860T>G were not reported previously. Considering the complexity of the genetics and phenotypes of the cystic renal diseases, genetic diagnosis is crucial to give accurate etiological diagnosis, which may benefit the clinic management.

OBJECTIVE: To carry out genetic analysis for a family with a fetus manifesting bilateral polycystic renal dysplasia and oligohydramnios at 16 gestational week and a previous history for fetal renal anomaly. METHODS: Ultrasound scan was carried out to detect the morphological changes. Following genetic counselling, the parents had decided to terminate the pregnancy. Fetal kidneys were subjected to histological examination. Target capture and next generation sequencing (NGS) was applied to the abortus to detect potential variants. The results were verified by Sanger sequencing. RESULTS: Histological examination of fetal kidneys revealed cystic changes without cortex, medulla or normal renal structure. NGS has identified a heterozygous c.100+1G>A variant and deletion of exon 3 of the INVS gene, which were respectively inherited from the mother and father. CONCLUSION Through NGS and Sanger sequencing, the fetus was diagnosed with type II nephronophthisis (NPHP2). Above result can provide guidance for further pregnancy and enforce understanding of clinical features and genetic etiologies for NPHP.
Female ; Fetus ; Genetic Testing ; Heterozygote ; Humans ; Mutation ; Polycystic Kidney, Autosomal Dominant ; diagnostic imaging ; genetics ; Pregnancy ; Sequence Deletion ; genetics ; Transcription Factors ; genetics ; Ultrasonography