[Exploration and thinking on functional preservation after orthotopic neobladder construction].

The concept of functional preservation after orthotopic neobladder construction has gradually attracted attention. Reconstruction of urine storage and voiding is the basic function preservation of orthotopic neobladder. Clinical exploration mainly focuses on the optimization of neobladder reconstruction methods and procedures, and there is still a lack of summary of existing surgical characteristics and high-quality functional comparative studies. For strictly selected patients, on the basis of tumor control and standardized postoperative rehabilitation guidance, most patients with preserved nerve can retain satisfied sexual function after surgery. The protection of neurovascular bundle and ancillary structures combined with postoperative exercise is crucial to the improvement of urinary continence. According to the characteristics of patients, choosing the appropriate urinary diversion methods and function preserving can help patients establish a normal life style after surgery and improve their self-image and quality of life.

[Risk factors of permanent stoma in rectal cancer patients undergoing transabdominal anterior resection with temporary stoma].

Objective: To investigate the risk factors of turning temporary stoma into permanent stoma in rectal cancer patients undergoing transabdominal anterior resection with temporary stoma. Methods: A case-control study was carried out. Data of rectal cancer patients who underwent transabdominal anterior resection with temporary stoma and completed follow-up in Department of General Surgery of Xiangya Hospital of Central South University from June 2008 to June 2018 were collected and analyzed. In this study, temporary stoma included defunctioning stoma (ostomy was made during operation) and salvage stoma (ostomy was made within one month after operation due to anastomotic leakage or severe complications). Cases of multiple intestinal tumors were excluded. A total of 308 rectal cancer patients were enrolled in the study, including 198 males and 110 females with a median age of 56 (48-65) years. Ninety-four patients received intraperitoneal chemotherapy during operation. Among 308 patients, upper rectal cancer was observed in 64 cases, middle rectal cancer in 89 cases and low rectal cancer in 155 cases. Twenty patients underwent transverse colostomy and 288 underwent ileostomy. Phone call following-up was conducted from August to September 2019 to investigate whether stoma was reversed, causes of reversal failure, and tumor relapsed or not in detail. Permanent stoma was defined as that the stoma was still not reversed by the latest follow-up. The univariate analysis was performed with chi-square test or Fisher's exact test, and variables with P value < 0.10 were included in the non-conditional logistic regression model for multivariate analysis. Results: The median follow-up time was 54.3 (32.4-73.8) months. During follow-up, 8 cases had local recurrence and 37 cases had distant metastasis. Among the 308 patients with temporary ostomy, 247 (80.2%) patients had stomas reversed and the median interval time was 4.5 (3.5-6.1) months. The median interval time in 65 patients with salvage stoma was significantly longer that in 182 patients with defunctioning stoma [5.5 (4.3-7.5) vs. 4.2 (3.4-5.5) months; Z=-4.387, P<0.001]. The temporary ostomy was confirmed to become permanent stoma in 61 patients (19.8%), including 45 cases of defunctioning stoma and 16 cases of salvage stoma. Univariate analysis showed that preoperative anemia, intraperitoneal chemotherapy during operation, middle rectal cancer, transverse colostomy, pathological stage, postoperative local recurrence and distant metastasis were associated with permanent stoma (all P<0.10). Multivariate analysis revealed that the intraperitoneal chemotherapy during operation (OR=1.961, 95% CI: 1.029-3.738, P=0.041), middle rectal cancer (OR=2.401, 95% CI: 1.195-4.826, P=0.014), transverse colostomy (OR=3.433, 95% CI: 1.234-9.553, P=0.018), and distant metastasis (OR=8.282, 95% CI:3.820-17.954, P<0.001) were independent risk factors of permanent stoma. Conclusions: There is high risk of turning temporary stoma into permanent stoma among rectal cancer patients undergoing transabdominal anterior resection who receive intraperitoneal chemotherapy during operation, present as the middle rectal cancer, undergo transverse colostomy or develop distant metastasis. Surgeons need to evaluate and balance the risks and benefits thoroughly, and then inform the patients in order to avoid potential conflicts.

Clinical significance of tumor mutation burden and DNA damage repair in advanced stage non-small cell lung cancer patients.

OBJECTIVE: This study aimed to investigate the impact of tumor mutational burden (TMB) and DNA damage repair (DDR) gene alteration on overall survival (OS) in advanced non-small cell lung cancer (NSCLC) patients. PATIENTS AND METHODS: A DNA library of cancer cells from 67 NSCLC patients in stages III-IV was constructed for next-generation sequencing (NGS). Geneseeq422 probes were used for hybridization enrichment. The target-enriched library was sequenced on HiSeqNGS platforms, and we analyzed the relevant signaling pathways. Then, we correlated the OS of the patients with TMB and DDR mutations. RESULTS: Many significant alterations were found, including in the EGFR, p53, KRAS, RB1, ERBB2, NF1, DNMT3A, ALK, MYC, PIK3CA, ROS1, BRAF, ARID1A, PTEN, CDKN2A, and FGF19 genes. We also identified many mutations in the genes relevant to the DDR pathway. Interestingly, we found that the TMB of patients with DDR gene mutations was dramatically higher than that in the DDR wild-type (WT). Univariable analysis showed that DNMT3A, RB1, DDR pathway-related gene mutations, and TMB were critical factors for the effects on OS. Multivariable analysis confirmed that DNMT3A and mutations in the DDR pathway-related genes were important for predicting OS. CONCLUSIONS: Multiple mutations in the genes of the DDR pathway caused higher TMB levels, which resulted in longer OS. By contrast, OS was significantly longer in patients with non-DNMT3A mutations than in those with DNMT3A variants. DNMT3A alteration in NSCLC patients led to poor outcomes.

[Interpretation of clinical practice guideline for anorectal day surgery 2019 edition].

As the rapid development of minimally invasive techniques, anesthesia, and enhanced recovery after surgery (ERAS), anorectal day surgery receiving more and more attention by improving efficiency of medical care while reducing cost and hospitalized infection. However, day surgery also faces the challenge of completing the whole process from patient admission to discharge within 24 hours. Therefore, establishing a reasonable and detailed day surgery process is the cornerstone to guarantee safe medical practice and patients satisfaction. National Clinical Research Center for Geriatric Disorders (Xiangya), together with China Ambulatory Surgery Alliance formulates the clinical practice guideline for anorectal day surgery 2019 edition. Here we make some interpretations of the guidelines on the detailed process of anorectal day surgery, including indication, preoperative examination, preoperative risk evaluation, health education, assessment of day surgery anesthesia and before leaving postanesthesia care unit (PACU), postoperative management, assessment of discharge and follow-up, for the convenience of various medical centers.

Downregulation of microRNA-383 is associated with male infertility and promotes testicular embryonal carcinoma cell proliferation by targeting IRF1.

Our previous studies have shown that microRNA-383 (miR-383) expression is downregulated in the testes of infertile men with maturation arrest (MA). However, the underlying mechanisms of miR-383 involved in the pathogenesis of MA remain unknown. In this study, we showed that downregulation of miR-383 was associated with hyperactive proliferation of germ cells in patients with mixed patterns of MA. Overexpression of miR-383 in NT2 (testicular embryonal carcinoma) cells resulted in suppression of proliferation, G1-phase arrest and induction of apoptosis, whereas silencing of miR-383 reversed these effects. The effects of miR-383 were mediated through targeting a tumor suppressor, interferon regulatory factor-1 (IRF1), and miR-383 was negatively correlated with IRF1 protein expression in vivo. miR-383 inhibited IRF1 by affecting its mRNA stability, which subsequently reduced the levels of the targets of IRF1, namely cyclin D1, CDK2 and p21. Downregulation of IRF1 or cyclin D1, but not that of CDK2, enhanced miR-383-mediated effects, whereas silencing of p21 partially inhibited the effects of miR-383. Moreover, miR-383 downregulated CDK4 by increasing proteasome-dependent degradation of CDK4, which in turn resulted in an inhibition of phosphorylated retinoblastoma protein (pRb) phosphorylation. These results suggest that miR-383 functions as a negative regulator of proliferation by targeting IRF1, in part, through inactivation of the pRb pathway. Abnormal testicular miR-383 expression may potentiate the connections between male infertility and testicular germ cell tumor.

Chemo- and radio-protective effects of polysaccharide of Spirulina platensis on hemopoietic system of mice and dogs.

AIM: To observe polysaccharide of Spirulina platensis (PSp) on the hematopoietic system of mouse and dogs which were damaged by injection of cyclophosphamide (CTX) and 60Co-gamma irradiation. METHODS: CTX and 60Co gamma ray were used to induce bone marrow damage, and the experimental animals were ig with different dose of PSp in vivo, after 12-d and 21-d administration, the whole blood cells and nucleated cells in bone marrow were measured, and the DNA in bone marrow were inspected by UV-spectrophotometer. RESULTS: CTX and 60Co-gamma irradiation induced hemopoietic system damage in mice and dogs, respectively. PSp 30, 60 mg/kg increased the level of the white cells in blood and nucleated cells and DNA in bone marrow in mice but had no effects on red cells and hemoglobins. PSp 12 mg/kg increased the level of red cells, white cells, and hemoglobins in blood and nucleated cells in bone marrow in dogs (P < 0.01), and the effects of PSp 60 mg/kg were better than that of berbamine hydrochloride 60 mg/kg. CONCLUSION: PSp has chemo-protective and radio-protective capability, and may be a potential adjunct to cancer therapy.

Transmission of porcine endogenous retroviruses in severe combined immunodeficient mice xenotransplanted with fetal porcine pancreatic cells.

BACKGROUND: Xenotransplantation using pig organs or tissues may alleviate the human donor organ shortage. However, one concern is the potential transmission of pig pathogens to humans, especially pig endogenous retroviruses (PERV), which infect human cell lines in vitro. In this report, the cross-species in vivo transmission of PERV by xenotransplantation was studied using a severe combined immunodeficient (SCID) mouse model. METHODS: Twenty-one SCID mice were transplanted with fetal pig pancreatic cells and left for periods from three to 41 weeks before being killed. DNA and RNA were extracted from liver, spleen, and brain of these mice, and examined for PERV using nested polymerase chain reaction (PCR) and reverse transcriptase-PCR. The pig mitochondrial cytochrome oxidase II subunit gene (COII) was also amplified to monitor the presence of pig cell microchimerism in xenotransplanted tissues, and a housekeeping gene was included to monitor the DNA quality and quantity. RESULTS: Examination of 39 DNA samples from tissues of the 21 xenografted mice identified two mouse tissues (M4-liver and M19-spleen) that were positive for PERV but negative for COII. A total of 23 (59%) of the mouse tissues were positive for both PERV and COII, 6 (16%) were negative for both, and 8 (20%) were positive for COII only. PCR and direct sequencing of the PCR products identified three PERV variants, which were different from the PERV sequence detected by PCR direct sequencing from the pig donor cells. CONCLUSIONS: The PERV+/COII- results from M4-liver and M19-spleen indicated the presence of PERV transmission from pig to mouse tissue. The PERV variants detected in the mouse tissues indicated that different PERVs were transmissible from the pig to mouse tissue during xenotransplantation. The negative reverse transcriptase-PCR results for PERV from three mouse samples including M4-liver and M19-spleen suggest there was no active PERV transcription in the mouse tissues, although this would need to be studied further.

Genetic analysis of regions involved in replication and cadmium resistance of the plasmid pND302 from Lactococcus lactis.

The 8.8-kb Lactococcus lactis plasmid pND302 encodes resistance to cadmium (CdR). Regions of pND302 involved in replication and CdR were subcloned and sequenced. The replication region is localized on a 1.5-kb region and consists of an open reading frame (repB) preceded by a noncoding AT-rich sequence (ori) which is highly homologous to lactococcal theta-type replicons. The CdR determinant is localized on a 2.9-kb region and encodes putative proteins similar to the Cd(2+)-specific P-type efflux ATPase (CadA) and the transcriptional regulatory repressor (CadC) identified in Staphylococcus aureus, Bacillus firmus, and Listeria monocytogenes. Similar CdR determinants were also detected by PCR in other CdR plasmids isolated from different L. lactis strains.

The effect of demineralized bone matrix on the healing of intramembranous bone grafts in rabbit skull defects.

A clinical dilemma exists regarding the type of bone that should be used to replace diseased or traumatized osseous tissue. Oral, plastic, and orthopedic surgeons normally implant viable mineralized endochondral (EC) autografts or demineralized EC allografts. A few clinicians have recognized the disadvantages of using EC bone in craniofacial surgery and advocated the replacement of intramembranous (IM) bone with healthy IM bone. However, controversy and uncertainty surround our understanding of these matrices to induce bone formation. Recent studies have advocated the use of other materials with osteoinductive properties, such as demineralized bone matrix (DBM). The proposed delivery system used in this study included IM bone grafts, DBM, and fixation of the IM bone graft. The purpose of this work was to gain further insights into the mechanism of healing of IM bone, in both the presence and the absence of DBM, and to compare the healing of IM bone grafts with that of DBM alone. Critical-sized (10 x 5 mm), full-thickness bony defects in rabbit parietal bone, devoid of periosteum, were filled with IM bone graft (mandible) alone, demineralized cortical bone matrix (DBM) alone, or combined DBM-IM bone graft, or were left unfilled. Histologic changes were examined 14 days later. The IM bone graft healed through IM ossification with no intermediate cartilage stage. DBM and composite DBM-IM healed through an EC ossification with an intermediate cartilage stage. It is hypothesized that the role of the IM graft is to induce neovascularization into the defect site, and that the undifferentiated mesenchymal cells in the perivascular region of the new blood vessels are induced by the bone morphogenetic protein(s) in the DBM into bone-forming cells.

Sister chromatid exchanges in lymphocytes of early cases of nasopharyngeal carcinoma and in 100 healthy subjects.

Sister chromatid exchange rate was studied in 12 early diagnosed cases of nasopharyngeal carcinoma and in their paired controls. Exchange frequencies were also analyzed in 100 healthy subjects distributed in four regions of Hunan Province and correlated to nationality, age and sex. The incidence of sister chromatid exchange was significantly higher in the cancer patients than in the normal controls. No correlation was found between the frequency of sister chromatid exchange and region, nationality, age or sex.