[Application of the two-step sentinel lymph node biopsy with double-tracer in early staged endometrial cancer].

Objective: To explore the feasibility and clinical value of sentinel lymph node (SLN) biopsy through cervix-uterine combined two-step injection with two tracers in patients with early stage endometrial cancer. Methods: From July 2019 to April 2021, a total of 73 patients, aged (54.2±3.3) year, who were preoperatively diagnosed as stage Ⅰ-Ⅱ endometrial cancer (including 56 low-risk patients and 17 medium-high risk patients) in Affiliated Hospital of Qingdao University were selected. According to the different sites of tracer injection, the patients were randomly divided into three groups: cervical injection group (25 cases): 1 ml of nano-carbon was used to inject at 3 and 9 o'clock in the cervix; uterine injection group (21 cases): the magnetic resonance imaging examination was performed to determine the location of the lesion, and 4 ml of methylene blue was injected into the uterine body at 2 sites where the lesion was located; combined injection group (27 cases): cervical injection of nano-carbon (1 ml) combined with uterine injection of methylene blue (4 ml). The SLN in all patients were identified under laparoscopy, removed, and followed by frozen pathological examination. Pathological ultra-staging was performed if the postoperative pathological outcome of SLN was negative. The total detection rate of SLN, bilateral pelvic SLN detection rate, sensitivity, negative predictive value, and location of SLN in each group were calculated and compared. Results: (1) In 73 patients with endometrial cancer, the overall detection rate of SLN was 88% (64/73), the detection rate of bilateral pelvic SLN was 67% (49/73), and the detection rate of para-aortic SLN was 49% (36/73). The overall detection rate of SLN (71%, 15/21) and bilateral pelvic SLN (43%, 9/21) in the intrauterine injection group were significantly lower than those in the cervical injection group [92% (23/25), 76% (19/25), respectively] and the combined injection group [96% (26/27), 78% (21/27), respectively; all P<0.05]; the detection rate of para-aortic SLN in the cervical injection group (28%, 7/25) was significantly lower than those in the intrauterine injection group and combined injection group [52% (11/21) and 67% (18/27), respectively; both P<0.05]. Among 73 cases with endometrial cancer, 9 had lymph node metastasis confirmed by postoperative pathological examination, 8 of them had lymph node metastasis detected by SLN and 1 had no lymph node metastasis detected by SLN, with a total sensitivity of 89% and a negative predictive value of 98%. The sensitivity and negative predictive value of cervical injection group and combined injection group were 100%, while the sensitivity and negative predictive value of intrauterine injection group were 67% and 95%. Among 56 low-risk patients, only one patient with lymph node metastasis was confirmed by postoperative pathology by SLN detection, and the metastasis rate was 2% (1/56), and the sensitivity and negative predictive value were 100%. Lymph node metastasis was confirmed in 8 of 17 patients (8/17) with a sensitivity of 88% and a negative predictive value of 90%. (2) A total of 459 SLN were detected in 73 endometrial cancer patients, with the highest proportion of external iliac (33.3%, 153/459).The obturator foramen was 25.3% (116/459), para-aortic 19.6% (90/459), iliac 12.0% (55/459), and presacral 9.8% (45/459). The proportion of para-aortic SLN in the cervical injection group was 12.4% (21/169), which were significantly lower than that in the intrauterine injection group and the combined injection group [27.4% (26/95) and 22.1% (43/195), respectively; both P<0.05]. (3) Pathological super-staging results: among 64 patients with negative SLN routine paraffin pathology, 4 cases of lymph node micro-metastases and 1 case of isolated tumor cell metastasis were detected, and the SLN micro-metastases rate was 8% (5/64), including 2 cases of low-risk patients and 3 cases of medium-high risk patients. Conclusions: SLN biopsy has high sensitivity and negative predictive value in patients with early endometrial cancer and could be used as an alternative to systematic lymph node dissection in low-risk patients. The SLN mapping through cervical-uterine combined injection could further improve the detection rate effectively and avoid the missed detection of positive para-aortic lymph node, especially for high-risk patients or patients with fundal tumor involvement.

Abnormal expression of Rap2A as a prognostic marker for human breast cancer.

OBJECTIVE: The aim of this study was to explore the expression of GTPase protein Ras-related protein Rap-2a (Rap2A) in breast cancer (BC). Furthermore, the associations of Rap2A with clinicopathological parameters of BC patients were investigated. PATIENTS AND METHODS: quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was used to examine Rap2A expression in BC tissues and cells. The association between Rap2A expression and clinicopathological characteristics was analyzed by Chi-square test. Low expression of Rap2A in BC cells was conducted by transfection of small interfering RNA (siRNA). Subsequently, colony formation assay and transwell assay were used to detect the proliferation and invasion abilities of BC cells, respectively. RESULTS: Rap2A was highly expressed in both BC tissues and cells (p<0.05). Further analysis showed that tumor size, clinical stage, and distant metastasis were correlated with Rap2A expression (p<0.05). Besides, inhibition of Rap2A significantly decreased the proliferation and invasion abilities of BC cells (p<0.05). CONCLUSIONS: Rap2A acted as a promotor in the development of BC. Our findings suggested that Rap2A might be a new potential therapeutic target marker for BC treatment.

[Clinical analysis of surgical treatment of post-infarction left ventricular aneurysm: a series of 254 patients].

Objective: To examine the efficacy of two surgical procedures on post-infarction left ventricular aneurysm. Methods: The clinic data of 254 patients with post-infarction left ventricular aneurysm, who underwent surgical ventricular reconstruction between January 1997 and December 2019 in Department of Cardiovascular Surgery, Nanjing Hospital Affiliated to Nanjing Medical University was analyzed retrospectively. There were 183 males and 71 females aged from 31 to 81 years, with a median age of 64.6 years. Based on the size of the ventricular aneurysm, there were 73 patient received linear reconstruction (linear group) and 181 patients received endoventricular patch plasty technique (patch plasty group). Ejection fraction, left ventricular systolic and end diastolic volume and left ventricular systolic and end diastolic volume index were recorded preoperatively, 2-week, 3-month, 1-year and 5-year after operation. The survival curves were plotted with Kaplan-Meier method and the survival rates were compared by Log-rank test. Results: All patients underwent surgery with a mean cardiopulmonay bypass duration of (92±32) minutes (44 to 196 minutes) and aortic cross clamp duration of (67±22) minutes (33 to 152 minutes).There were 9 perioperative deaths with a mortality rate of 3.5%. Angina pectoris of other cases are relief and heart function improved greatly. Five years after operation, the percentage of cardiac function (New Yord Heart Association) class Ⅲ to Ⅳ patients decreased from 96.1%(244/254) to 9.9%(16/161). There was no significant difference in survival rate between linear group and patch plasty group at 1-, 3-, 5-years postoperatively (96%, 91%, 77% vs. 96%, 90%, 79%, P=0.562). Ejection fraction increased from (39±10)% (range: 22% to 50%) preoperatively to (46±6)% (range: 39% to 54%) 1-year postoperatively in the linear group, while increased from (38±13)% (range: 26% to 51%) preoperatively to (50±6)% (range: 39% to 55%) in the patch plasty group. Conclusions: Left ventricular reconstruction is quite effective for patients with post-infarction left ventricular aneurysm. The choice of operative approaches is determined by the size and range of ventricular aneurysm. Both linear reconstruction and endoventricular patch plasty technique can got similarly surgical outcomes with near and late curative effect.

[Application effects of enhanced computed tomography and three-dimensional reconstruction technology in reconstruction of pediatric post-burn scars with expanded flaps].

Objective: To explore the application effects of enhanced computed tomography (CT) and three-dimensional reconstruction technology in the reconstruction of pediatric post-burn scars with expanded flaps. Methods: From May 2016 to March 2019, 19 children with hypertrophic scars after thermal injuries were admitted to our unit, including 10 boys and 9 girls, aged from 4 years and 5 months to 15 years and 11 months. The area of scars ranged from 5 cm×4 cm to 23 cm×9 cm. One or more skin and soft tissue expanders with suitable volume and shape were implanted into the normal skin area around scar of children. Three to six months later, enhanced CT and three-dimensional reconstruction were performed before the second stage operation to obtain three-dimensional images of the vascular branches in the donor site for expanded flaps to be cut, so as to determine the course and distribution of the vascular branches and guide the design of expanded flaps. According to the design scheme, the resection of scar, removal of expanders, and excision and transfer of flaps were performed to repair the wounds after scar resection. The area of flaps ranged from 6 cm×4 cm to 25 cm×10 cm. The donor site was closed directly. The number of flaps was counted. The anatomical structure, vascular distribution, and adverse reactions during enhanced CT and three-dimensional reconstruction of site for expanded flaps to be cut, the survival of expanded flaps and the follow-up after the second-stage operation were observed. Results: A total of 48 expanded flaps were designed and excised in 19 children. The anatomical structure of the site for expanded flaps to be cut and the adjacent spatial position relationship were visually observed through the three-dimensional reconstruction after enhanced CT, and no adverse reactions were observed. Arterial branch blood supply or venous return was observed in 29 sites for expanded flaps to be cut. All the expanded flaps survived well without blood supply disorder after the second stage operation. The children were followed up for 6 months to 1 year and 6 months after the second stage operation. The appearance of the flaps was natural, and the color and thickness of the flaps were similar to those of the surrounding normal skin, except for one child with obvious linear scar. Conclusions: Enhanced CT and three-dimensional reconstruction can assist the vascular assessment of the expended flaps, which is helpful for rational design of the flap excision and transfer protocol to improve the survival rate of flaps. Thus, it has certain clinical application value in the reconstruction of post-burn scar in children with expanded flaps.

Euxanthone exhibits anti-proliferative and anti-invasive activities in hepatocellular carcinoma by inducing pyroptosis: preliminary results.

OBJECTIVE: Hepatocellular carcinoma (HCC) is one of the most common causes of cancer-related deaths. Euxanthone, a xanthone compound extracted from Polygala caudata, possesses a variety of pharmacological activities. This work aimed to explore whether euxanthone exhibits anti-cancer activities in HCC. PATIENTS AND METHODS: Tissue specimen was collected and the mRNA and protein expression of marker proteins were detected by qRT-PCR and Western blot, respectively. The viable cell number was assessed by CCK-8 assay. TUNEL assay was utilized to determine cell death. Cell migration was measured by Wound healing. Cell invasion was measured by transwell assay. Xenograft model was established to evaluate the efficacy of euxanthone in vivo. RESULTS: We demonstrated that HCC tissue and HCC cell line presented with lower expression of caspase-1, IL-1ß, and IL-18. Euxanthone markedly suppressed the proliferation of HCC cells and induced cell death. In addition, euxanthone inhibited cell migration and invasion. Meanwhile, our results showed that euxanthone promoted cell death in a caspase-2 dependent manner. In vivo data also showed that euxanthone suppressed tumor growth and promoted pyroptotic cell death. CONCLUSIONS: We showed that that euxanthone may be a candidate of anticancer agent and provide clinical benefits for patients with HCC.

Doublets versus single-agent therapy as first-line therapy for elderly patients with advanced non-small cell lung cancer? A systematic review of randomised controlled trials.

INTRODUCTION: Although the standard treatment for advanced non-small cell lung cancer (NSCLC) patients is platinum-based doublet chemotherapy, single-agent therapy is still preferred in elderly patients. Comparison of the efficacy of various combinations of doublets with single-agent chemotherapy is somehow contradictory. This study conducted a systematic review to evaluate the efficacy and tolerability of the third-generation agent-based doublets vs. single-agent chemotherapy in elderly NSCLC patients. METHODS: Electronic (PubMed, EMBASE and Cochrane Library database) and manual searches were conducted to collect data from published, randomised, phase 2 and 3 trials which compared doublets with a third-generation single-agent chemotherapy in elderly patients. Pooled relative risks (RRs) were calculated for the incidences of overall response rate (ORR), 1-year survival rate (1-y SR), and grade 3/4 toxicities. RESULTS: Seven eligible trials (2219 patients) were selected from 1170 studies that were initially identified. A significant difference in ORR favouring doublets over single agents was observed [RR, 1.59; 95% confidence interval (95% CI), 1.36-1.86; p < 0.0001] with a slightly, but not significantly improved 1-y SR (RR, 1.19; 95% CI, 0.98-1.45, p = 0.007). Subgroup analysis suggested that platinum (RR, 1.94; 95% CI, 1.47-2.55, p < 0.0001) or non-platinum- (RR, 1.45; 95% CI, 1.20-1.75, p < 0.0001) based doublets could improve ORR, and the grade 3/4 thrombocytopaenia (RR, 6.64; 95% CI, 1.78-24.86, p = 0.005) and anaemia (RR, 2.86; 95% CI, 1.62-5.05, p < 0.0001) were preferred to occur in platinum-based doublets. CONCLUSIONS: Doublets appear to be more effective and tolerable than single-agent therapy for treating elderly advanced NSCLC patients, and therefore could be considered as a treatment option for elderly populations with good physical status.

SecA is required for the insertion of inner membrane proteins targeted by the Escherichia coli signal recognition particle.

Recent work has demonstrated that the signal recognition particle (SRP) is required for the efficient insertion of many proteins into the Escherichia coli inner membrane (IM). Based on an analogy to eukaryotic SRP, it is likely that bacterial SRP binds to inner membrane proteins (IMPs) co-translationally and then targets them to protein transport channels ("translocons"). Here we present evidence that SecA, which has previously been shown to facilitate the export of proteins targeted in a post-translational fashion, is also required for the membrane insertion of proteins targeted by SRP. The introduction of SecA mutations into strains that have modest SRP deficiencies produced a synthetic lethal effect, suggesting that SecA and SRP might function in the same biochemical pathway. Consistent with this explanation, depletion of SecA by inactivating a temperature-sensitive amber suppressor in a secAam strain completely blocked the membrane insertion of AcrB, a protein that is targeted by SRP. In the absence of substantial SecA, pulse-labeled AcrB was retained in the cytoplasm even after a prolonged chase period and was eventually degraded. Although protein export was also severely impaired by SecA depletion, the observation that more than 20% of the OmpA molecules were translocated properly showed that translocons were still active. Taken together, these results imply that SecA plays a much broader role in the transport of proteins across the E. coli IM than has been previously recognized.

Mutation in Cys662 of Escherichia coli DNA topoisomerase I confers temperature sensitivity and change in DNA cleavage selectivity.

Cys662 is one of the 12 cysteine residues proposed to be co-ordination sites for binding of three Zn(II) in Escherichia coli DNA topoisomerase I. Oligonucleotide-directed mutagenesis was used to convert Cys662 to either serine or histidine. The mutant enzymes were overexpressed and purified to homogeneity. Analysis of the purified enzymes demonstrated that the mutations resulted in loss of one tightly bound Zn(II). In vivo complementation tests and in vitro relaxation activity assays at different temperatures showed that the partial relaxation activities retained in the two mutant enzymes were temperature sensitive. Fluorescence measurements indicated that the wild-type and mutant enzymes have structural differences. When DNA cleavage specificity was examined, the mutant enzymes were found to have altered cleavage site preferences. The preferred cleavage sites of the wild-type enzyme all had a cytosine residue four nucleotides to the minus side of the break. The cleavage sites produced by the mutant enzymes did not show a preference for cytosine at that position.