[Regarding the selection of individualized therapy after neoadjuvant therapy for gastrointestinal tumors].

Gastrointestinal tumors have been widely concerned because of increasing morbidity and mortality. In the process of exploring the therapeutic patterns of gastrointestinal tumors, patients treated with neoadjuvant therapies have good effect of tumor regression and favorable prognosis. Thus, neoadjuvant therapy strategies are recommended by major guidelines of gastrointestinal tumors in the world. Meanwhile, they have a great impact on the traditional methods of surgery, the influence mainly involves the reduction of the surgical margin and the scope of lymph node dissection in gastric cancer, while involves performing organ preservation and watch & wait in selective patients with colorectal cancer. These effects and changes were based on effective control of local recurrence by neoadjuvant therapies, and the advantages of neoadjuvant therapy in terms of tumor regression and survival supported by many studies. It is also based on the patient's desire for organ preservation and non-surgical treatment. Meanwhile, application of neoadjuvant therapy strategies increase surgical difficulty and postoperative complications, but the overall impact on patient prognosis is weak. Therefore, the selection of an appropriate treatment model after neoadjuvant therapy requires an effective overall post-treatment evaluation. In particular, it is necessary to pay attention to the evaluation of imaging, endoscopy, etc., while effectively performing monitoring and follow-up, and finally establishing an appropriate salvage treatment. This article will review the status and problems of individualized treatment after neoadjuvant therapy of gastrointestinal tumor.

[Preclinical study of T cell receptor specifically reactive with KRAS G12V mutation in the treatment of malignant tumors].

OBJECTIVE: KRAS gene is one of the most common mutations of proto-oncogenes in human tumors, G12V is one of the most common mutation types for KRAS. It's challenging to chemically acquire the targeted drug for this mutation. Recent studies reported that this mutation peptides can form a neoepitope for T cell recognition. Our study aims to clone the T cell receptor (TCR) which specifically recognizes the neoepitope for KRAS G12V mutation and constructs TCR engineered T cells (TCR-T), and to investigate if TCR-Ts have strong antitumor response ability. METHODS: In this study, tumor infiltrating lymphocytes were obtained from one colorectal cancer patient carrying KRAS G12V mutation. Tumor-reactive TCR was obtained by single-cell RT-5' rapid-amplification of cDNA ends PCR analysis and introduced into peripheral blood lymphocytes to generate TCR-Ts. RESULTS: We obtained a high-affinity TCR sequence that specifically recognized the HLA-A*11:01-restricted KRAS G12V8-16 epitope: KVA11-01. KVA11-01 TCR-T could significantly kill various tumor cells such as PANC-1, SW480 and HeLa (overexpressing HLA-A*11:01 and KRAS G12V), and secreting high levels of interferon-γ (IFN-γ). Non-specific killing experiments suggested KVA11-01 specifically recognized tumor cells expressing both mutant KRAS G12V and HLA-A*11:01. In vivo assay, tumor inhibition experiments demonstrated that infusion of approximately 1E7 KVA11-01 TCR-T could significantly inhibit the growth of subcuta-neously transplanted tumors of PANC-1 and HeLa (overexpressing HLA-A*11:01 and KRAS G12V) cells in nude mice. No destruction of the morphologies of the liver, spleen and brain were observed. We also found that KVA11-01 TCR-T could significantly infiltrate into tumor tissue and had a better homing ability. CONCLUSION: KVA11-01 TCR-T cells can effectively target a variety of malignant tumor cells carrying KRAS G12V mutation through in vitro and in vivo assay. KVA11-01 TCR-T cells have excellent biological activity, high specificity of target antigen and homing ability into solid tumor tissue. KVA11-01 TCR-T is expected to be an effective treatment for patients with KRAS G12V mutant solid malignancies.

[Clinical characteristics and vaccination status of SARS-CoV-2 Omicron variant infected children].

Objective: To investigate the clinical characteristics and vaccination status of SARS-CoV-2 Omicron variant infected children. Methods: A total of 105 children infected with Omicron variant admitted to Tianjin Haihe Hospital (designated referral hospital for SARS-CoV-2 infection in Tianjin) from January 8, 2022 to February 3 were included for a retrospective study. The cases were divided into pneumonia group and non-pneumonia group according to chest imaging. Based on the doses of inactivated SARS-CoV-2 vaccine, the children who completed SARS-CoV-2 antibody test within 3 days after hospitalization were divided into 2 dose group and<2 dose group.Rank sum test and Chi-square test were used for the comparison between the groups. Results: The age of these 105 children was 10 (8, 11) years on admission, 53 children were males and 52 were females. Eighty-seven cases (82.9%) had mild symptoms, 13 cases (12.4%) had common symptoms and 5 cases (4.8%) were asymptomatic. Ninety-one cases (86.7%) completed 2 doses vaccination. The clinical symptoms were characterized by cough (74 cases, 70.5%), fever (58 cases, 55.2%), sore or dry throat (34 cases, 32.4%), nasal congestion (28 cases, 26.7%), rhinorrhea (23 cases, 21.9%). None of the children received antivirals, steroids, immunosuppressant or oxygen therapy. Seventy-six cases(72.4%) received traditional Chinese medicine treatment. The pneumonia group had a higher rate of positive SARS-CoV-2 IgG within 1 day after admission (13/13 vs. 87.0% (80/92), χ2=42.81, P<0.001) than the non-pneumonia group. Among the 62 children who had serial SARS-CoV-2 antibody tests within 3 days after hospitalization, Compared to the<2 dose group, the 2 dose group had a higher rate of nucleic acid conversion within 16 days after onset and a higher rate of positive SARS-CoV-2 IgG 1 day after admission and 3 days after hospitalization (96.4% (54/56) vs. 4/6, 100.0% (56/56) vs. 2/6, 100.0% (56/56) vs. 3/6, all P<0.05). Conclusions: Most children infected with Omicron variant have mild symptoms, mainly respiratory infection symptoms. The proportion of SARS-CoV-2 antibody IgG positive in children who have received 2 doses of inactivated SARS-CoV-2 vaccines is higher,and the time of whose nucleic acid conversion may be shortened.

[Detection of SMO gene mutations in odontogenic keratocyst].

Objective: To detect the SMO mutations in odontogenic keratocyst (OKC) and to explore the mechanism behind. Methods: Patients with OKC who received treatment in the Department of Oral and Maxillofacial Surgery, School and Hospital of Stomatology,Peking University, from September 2012 to June 2017 were enrolled. OKC samples from 10 patients diagnosed as naevoid basal cell carcinoma syndrome (NBCCS)-related OKC (4 females and 6 males) and 20 patients diagnosed as sporadic OKC (7 females and 13 males) were collected. Genomic DNAs were extracted from fibrous capsules and epithelial lining respectively. SMO mutations were detected and analyzed by Sanger sequencing. Results: Three SMO mutations were found in one NBCCS-associated OKC who carrying c.2081C>G (p.P694R) mutation) and two sporadic OKC who carrying c.907C>T (p.L303F) mutation and c.1247_1248delinsAA (p.G416E), respectively), among which the first two mutations were novel mutations that had not been reported before. Besides, two mutations in sporadic OKC were not paired with PTCH1 mutations. Conclusions: In addition to PTCH1 gene mutations, SMO gene mutations also exist in OKC which might be related to the development of OKC.

[Clinical characteristics and prognostic analysis of double primary breast cancer and thyroid cancer].

Objective: To explore and describe clinicopathological characteristics and prognosis of patients with double primary breast cancer (BC) and thyroid cancer (TC). Methods: Medical records of 98 patients diagnosed with double primary breast and thyroid cancer in National Cancer Center (NCC)/Cancer Hospital between January 1, 2001 and December 31, 2020 were retrospectively collected. All of the patients were followed up until January 1, 2021 to acquire survival data. Univariate survival analysis was conducted by Kaplan-Meier method, and multivariate survival analysis was carried out using the Cox proportional hazard model. Results: All of 98 patients in the group were women. The age at diagnosis of the first tumor ranged from 26-72 years old, and the median age was 47 years old. The BC recurring TC (breast methyl) group included 18 cases, TC recurring BC (methyl breast) group included 60 cases, BC and TC simultaneously occurred group (the two are diagnosed within 3 months) included 20 cases. There were statistically significant differences in breast cancer pathological grading, breast cancer postoperative radiotherapy, and combined with other tumors in breast methyl group, methyl breast group and the simultaneous group (P<0.05). Among the 98 patients, 14 had recurrence and metastasis, and 7 died. The patients who died from tumors were all those with TC recurrence of BC. There were no statistically significant differences in the death, recurrence and metastasis of patients in the breast methyl group, methyl breast group and the simultaneous group (P>0.05). Univariate analysis showed that BC stage and estrogen receptor (ER) were related to overall survival (P<0.05), while the family history of BC, BC stage, and ER were not related with the recurrence and metastasis (P<0.05). Multivariate analysis showed that BC family history, ER positive, and the order of tumor diagnosis (TC recurring BC) were independent influencing factors for the recurrence and metastasis (P<0.05). Conclusion: ER negative is a poor prognostic factor for the double primary breast and thyroid cancer.

[Effects of tumor protein translation control antisense RNA1 on radiosensitivity, proliferation, migration and invasion of hepatocellular carcinoma cells by targeting miR-30c-5p].

Objective: To investigate the effects of tumor protein translation control antisense RNA1 (TPT1-AS1) on the radiosensitivity, cell proliferation, migration and invasion of hepatocellular carcinoma cells by targeting microRNA-30c-5p (miR-30c-5p). Methods: Thirty-four cases of liver cancer tissues and adjacent normal tissues were derived from liver cancer patients who were admitted to Shanxi Provincial People's Hospital from March 2016 to March 2018. Liver cancer HepG2 cell was transfected with negative control siRNA (si-NC group), si-TPT1-AS1 (si-TPT1-AS1 group), pcDNA3.1 (pcDNA3.1 group), pcDNA3.1-TPT1-AS1 (pcDNA3.1-TPT1-AS1 group), si-TPT1-AS1 and anti-miR-NC (si-TPT1-AS1+ anti-miR-NC group), si-TPT1-AS1 and anti-miR-30c-5p (si-TPT1-AS1+ anti-miR-30c-5p group), respectively. Real-time quantitative reverse transcription polymerase chain reaction (qPCR) was used to detect the transcription levels of TPT1-AS1 and miR-30c-5p in normal tissues adjacent to cancer and liver cancer tissues, the clone formation test was used to test the radiosensitivity of HepG2 cells, and the Methyl Thiazolyl Tetrazolium (MTT) test was used to test the proliferation of HepG2 cells. Cell cycle distribution was detected by flow cytometry, Transwell array was used to detect the migration and invasion ability of HepG2 cells, dual luciferase reporter array was used to verify the targeting relationship of TPT1-AS1 and miR-30c-5p, western blot was used to detect the expressions of proliferation, migration and invasion-related proteins. Results: The expression levels of TPT1-AS1 and miR-30c-5p in liver cancer tissues were 0.84±0.08 and 0.13±0.01, statistically different from 0.31±0.03 and 0.50±0.05 in normal tissues adjacent to cancer (P<0.05). When the cells were treated with 2, 4, 6, 8 Gy irradiation, the cell survival scores of the si-TPT1-AS1 group were 0.280±0.040, 0.069±0.011, 0.020±0.003 and 0.005±0.001, respectively, lower than 0.648±0.070, 0.348±0.080, 0.130±0.020 and 0.060±0.009 of the si-NC group (P<0.05), the radiosensitization ratio of the si-TPT1-AS1 group was 1.672. The number of cell migration and invasion in the si-TPT1-AS1 group were (50.00±4.36) and (44.00±4.03), respectively, which were lower than (109.00±8.68) and (94.00±7.49) in the si-NC group (P<0.05), the cell absorbance (A) values at 24, 48 and 72 hours were 0.28±0.03, 0.43±0.04 and 0.68±0.07, respectively, lower than 0.46±0.04, 0.87±0.08 and 1.35±0.13 of the si-NC group (P<0.05), the protein expression levels of Cyclin D1, p21, E-cadherin and MMP-2 were 0.25±0.02, 0.65±0.06, 0.68±0.07 and 0.27±0.03, respectively, statistically different from 0.88±0.08, 0.17±0.02, 0.14±0.01 and 0.89±0.09 of si-NC group (P<0.05), the proportions of S phase and G(2) phase in the si-TPT1-AS1 group were (17.82±1.03)% and (34.15±2.29)%, respectively, significantly different from (35.14±2.61)% and (16.84±1.21)% in the si-NC group (P<0.05). The luciferase activity of cells in the WT-TPT1-AS1+ miR-30c-5p group was 0.26±0.02, lower than 0.92±0.09 in the WT-TPT1-AS1+ miR-NC group (P<0.05). The cell survival scores in the si-TPT1-AS1+ anti-miR-30c-5p group were 0.450±0.081, 0.200+ 0.045, 0.070±0.010, 0.026±0.004 after treatment with 2, 4, 6, 8 Gy irradiation, higher than 0.285±0.043, 0.075±0.014, 0.028±0.004, 0.006±0.001 of si-TPT1-AS1+ anti-miR-NC group (P<0.05). The radiosensitization ratio of the si-TPT1-AS1+ anti-miR-30c-5p group was 0.694. The number of migration and invasion in the si-TPT1-AS1+ anti-miR-30c-5p group were 79.00±6.65 and 68.00±6.33, higher than (52.00±4.41) and (46.00±4.06) of si-TPT1-AS1+ anti-miR-NC Group (P<0.05), the A values at 24, 48 and 72 hours were 0.37±0.03, 0.64±0.06 and 0.96±0.09, respectively, higher than 0.26±0.03, 0.41±0.04, and 0.65±0.06 of si-TPT1-AS1+ anti-miR-NC group (P<0.05), the expression levels of Cyclin D1, p21, E-cadherin and MMP-2 protein were 0.57±0.06, 0.43±0.04, 0.43±0.04 and 0.64±0.06, statistically different from 0.24±0.02, 0.66±0.06, 0.65±0.06 and 0.28±0.03 of the si-TPT1-AS1+ anti-miR-NC group (P<0.05). Conclusions: The expression of TPT1-AS1 up-regulates in the liver cancer tissues. TPT1-AS1 may down-regulate miR-30c-5p expression, reduce the radiosensitivity of liver cancer cells, and promote the proliferation, migration and invasion of liver cancer cells.

[Exploration of the high-risk factors of catheter-related thrombosis in breast cancer].

Objective: To explore the high risk factors of catheter-related thrombosis (CRT) in breast cancer patients, and provide the basis for the development of appropriate prevention and treatment strategies. Methods: A total of 1 432 breast cancer patients scheduled to receive central venous catheterization in National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College from January 1, 2015 to August 31, 2019 were selected. Baseline information and catheterization information of patients were collected. The occurrence of CRT was confirmed by vascular ultrasound examination, and the influencing factors of CRT were analyzed. Results: The total number of catheter days were 121, 980 days in 1 432 patients with breast cancer, and the average number of catheter days in each patient was 85.2 days. The incidence of CRT was 6.8% (97/1 432), which was 0.79 cases/1 000 catheter days. Among 815 patients with centrally inserted central venous catheters (CICC), 43 (5.3%) had CRT, which was 0.70 cases/1 000 catheter days. Among 617 patients with peripherally inserted central venous catheters (PICC), 54 (8.8%) developed CRT, which was 0.90 cases/1 000 catheter days. CRT was most common in subclavian vein (63.9%). Multivariate regression analysis showed that age ≥ 60 years old (OR=1.712, 95% CI: 1.056-2.775, P=0.029), PICC (OR=1.732, 95% CI: 1.130-2.656, P=0.012), the catheter position except subclavian vein (OR=10.420, 95% CI: 1.207-89.991), secondary adjustment of catheter position (OR=3.985, 95% CI: 1.510-10.521, P=0.005) and high D-Dimer level (OR=1.129, 95% CI: 1.026-1.241, P=0.012)were independent risk factors for CRT. Conclusions: The CRT problem can't be ignored in the clinical treatment of breast cancer patients with central venous catheterization. Screening the appropriate age of patients and the type of central venous catheters, reducing the secondary adjustment of catheter position, and timely monitoring the level of D-dimer are helpful to the prevention and treatment of CRT.

Hypomethylating agents for elderly patients with acute myeloid leukemia: a PRISMA systematic review and meta-analysis.

OBJECTIVE: Abnormal DNA methylation plays a critical role in acute myeloid leukemia (AML) pathogenesis and hypomethylating agents (HMAs) such as decitabine (5-aza-29-deoxycytidine) and azacitidine (5-azacytidine) are considered efficacious for treating AML. This study aimed to identify if HMAs have therapeutic advantages compared with conventional care regimens (CCR) or placebo in elderly AML patients. MATERIALS AND METHODS: We systematically searched PubMed, Embase, and Cochrane Central Register of Controlled Trials from inception to November July 15, 2020. Randomized controlled trials that compared the efficacy and adverse events associated with HMAs, CCR, or placebo were searched. RevMan 5.3 software was used to calculate the hazard ratio (HR) and risk ratio (RR) with a 95% confidence interval (CI). RESULTS: Seven trials with a total of 1966 participants were included. Meta-analyses showed that the overall survival of HMAs was better than that of CCR [HR=0.76, 95% CI (0.69-0.85), (p<0.01)], and the complete remission rate of elderly AML patients was increased by HMAs compared with CCR [RR=1.46, 95%CI (1.08-1.99), p=0.01)]. HMA treatment showed higher incidence of neutropenia [RR=1.30 (95%CI 1.07-1.59, p=0.008)], thrombocytopenia [RR=1.14 (95%CI 1.01-1.59, p=0.04)], and pneumonia [RR=1.37 (95%CI 1.06-1.76, p=0.02)] compared with CCR. CONCLUSIONS: Although HMAs cause a higher incidence of adverse events such as neutropenia, thrombocytopenia, and pneumonia, demethylation drugs are well-tolerated and effective for treating AML in the elderly.

Integrative sequencing discovers an ATF1-motif enriched molecular signature that differentiates hyalinizing clear cell carcinoma from mucoepidemoid carcinoma.

OBJECTIVES: Salivary gland tumors are comprised of a diverse group of malignancies with widely varying prognoses. These cancers can be difficult to differentiate, especially in cases with limited potential for immunohistochemistry (IHC)-based characterization. Here, we sought to define the molecular profile of a rare salivary gland cancer called hyalinizing clear cell carcinoma (HCCC), and identify a molecular gene signature capable of distinguishing between HCCC and the histopathologically similar disease, mucoepidermoid carcinoma (MEC). MATERIALS AND METHODS: We performed the first integrated full characterization of five independent HCCC cases. RESULTS: We discovered insulin-like growth factor alterations and aberrant IGF2 and/or IGF1R expression in HCCC tumors, suggesting a potential dependence on this pathway. Further, we identified a 354 gene signature that differentiated HCCC from MEC, and was significantly enriched for genes with an ATF1 binding motif in their promoters, supporting a transcriptional pathogenic mechanism of the characteristic EWSR1-ATF1 fusion found in these tumors. Of the differentially expressed genes, IGF1R, SGK1 and SGK3 were found to be elevated in the HCCCs relative to MECs. Finally, analysis of immune checkpoints and subsequent IHC demonstrated that CXCR4 protein was elevated in several of the HCCC cases. CONCLUSION: Collectively, our data identify an ATF1-motif enriched gene signature that may have clinical utility for molecular differentiation of HCCCs from other salivary gland tumors and discover potential actionable alterations that may benefit the clinical care of recurrent HCCC patients.

Kaempferol suppresses proliferation and induces apoptosis and DNA damage in human gallbladder cancer cells through the CDK4/CDK6/cyclin D1 pathway.

OBJECTIVE: Kaempferol has been reported to play an anti-tumor role in various human cancers, while its role in gallbladder cancer (GBC) is unclear. MATERIALS AND METHODS: We found that kaempferol significantly inhibited the growth, invasion and migration, meanwhile induced apoptosis through cells arrested at G0/G1 phase of GBC cell lines, including GBC-SD and SGC996 cells in vitro. RESULTS: Kaempferol promoted the release of cytochrome C from the mitochondria to cytoplasm, the activation of c-caspase-3 and c-caspase-9 and increased the expression levels of pro-apoptotic factor Bax, meanwhile decreased the expression levels of anti-apoptotic factor Bcl-2. In addition, the expression levels of CDK4, CDK6 and cyclin D1, which are members of the CDK4/CDK6/cyclin D1 signaling pathway, were also decreased by kaempferol. Moreover, kaempferol could efficiently prevent tumor progression of GBC in the xenograft in vivo. CONCLUSIONS: Our results demonstrated that kaempferol suppressed GBC progression through activation of the CDK4/CDK6/cyclin D1 signaling pathway, suggesting that it might be a potential anti-tumor agent for clinical treatment of GBC.

Generalized temperature dependence model for anammox process kinetics.

Temperature is a key operational factor influencing the anammox process kinetics. In particular, at temperatures below 15 °C, the specific anammox activity (SAA) considerably decreases. This study aimed to describe the temperature dependence of the anammox process kinetics in the temperature range from 10 to 55 °C, including the specific characteristics of "cold anammox". The commonly used Arrhenius and extended and modified Ratkowsky equations were examined. The Ratkowsky equations yielded a strong correlation (coefficient of determination, R2 = 0.93-0.96) between the measured and predicted data over the analyzed temperature range (10-55 °C). However, these equations could not correctly reflect the anammox temperature dependence at temperatures below 15 °C (R2 = 0.36-0.48). Therefore, a new generalized temperature model was proposed. The generalized temperature equation (GTE) considered the division of the analyzed temperature range into three temperature ranges: 10-15 °C, 15-35 °C and 35-55 °C. The ranges correspond to "cold anammox", "(low) mesophilic anammox" and "thermophilic anammox". The applied approach yielded a strong correlation between the measured and predicted SAA (R2 = 0.97) over the temperature range from 10 to 55 °C and over the low-temperature range from 10 to 15 °C (R2 = 0.99). Overall, the GTE could enhance the predictions of the temperature dependence of the anammox process kinetics. The GTE can help examine anammox-based bioaugmentation systems operating at both high temperatures (sidestream reactors) and low temperatures (mainstream reactors).

[The value of conventional magnetic resonance imaging based radiomic model in predicting the texture of pituitary macroadenoma].

Objective: To investigate the value of conventional magnetic resonance imaging (MRI) based radiomic model in predicting the texture of pituitary macroadenoma. Methods: The complete data of 101 patients with pituitary macroadenoma confirmed by surgery and pathology in Yijishan Hospital of Wannan Medical College from December 2014 to December 2019 were retrospectively analyzed. According to the texture of the intraoperative pituitary tumor, patients were divided into soft group (n=58) and hard group (n=43). They were randomly divided into training group (n=72) and validation group (n=29) at a ratio of 7∶3. All patients underwent conventional MRI scan of the pituitary gland. Itk-snap software was used to manually outline the T(1)-weighted image (T(1)WI), T(2)-weighted image (T(2)WI) and enhanced T(1)WI image section by section on tumor area of interest (ROI) and perform three-dimensional fusion. Then AK software was imported to extract texture features. The regression analysis methods of minimum redundancy maximum relevance (mRMR) and least absolute shrinkage and selection operator (LASSO) were used for feature selection and radiomic signature establishment. The reliability of the model was verified by 100 leave-group-out cross validation (LGOCV), and the predictive ability of the model was evaluated by drawing the receiver operating characteristic (ROC) curve. The decision curve analysis (DCA) was used to evaluate the clinical application value of the model. Results: The AUC (Area Under the ROC Curve) (95%CI) values of T1WI, T2WI, enhanced T1WI, and the combined sequence model to predict the texture of pituitary macroadenomas in the training and validation groups were 0.91 (0.84-0.98) and 0.90 (0.78-1.00), 0.86 (0.78-0.95) and 0.83 (0.64-1.00), 0.90 (0.83-0.97) and 0.89 (0.77-1.00),0.92 (0.85-0.98) and 0.91 (0.79-1.00), respectively. DCA demonstrated that T(1)WI, T(2)WI, enhanced T(1)WI, and combined sequence model all had good net benefits in clinical practice. Conclusions: T(1)WI, T(2)WI, enhanced T(1)WI, and combined sequence model of conventional MRI all had high efficacy in predicting the texture of pituitary macroadenoma, which provided a new quantitative method for predicting the texture of pituitary macroadenoma.

[Protective effect and mechanism of mild hypothermia on liver injury after cardiopulmonary resuscitation in pigs].

Objective: To investigate the effect of mild hypothermia therapy on liver after cardiopulmonary resuscitation. Methods: Thirty-three inbred Chinese Wuzhishan (WZS) minipigs, weighted (28±2) kg, were used to establish a ventricular fibrillation model. And 30 animals survived after cardiopulmonary resuscitation reached basic life support. The surviving animals were randomly divided into two groups: mild hypothermia group (group M, n=15) and conventional treatment group (group C, n=15). All the animals were observed for 24 hours. Blood samples were extracted at baseline, 0.5, 1, 2, 4, 6, 12 and 24 h after successful resuscitation. The concentrations of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected at the time points. The enzyme-linked immunosorbent assay (ELISA) was used to detect the concentrations of interleukin-6 (IL-6) and tumour necrosis factor-alpha (TNF-α). The data were compared between the two groups, LSD test was used when the variance was homogeneous, and Tamhane T2 test was used when the variance was uneven. Results: Eleven pigs (73.3%) in the group M and 8(53.3%) in the group C survived at 24 h after successful resuscitation, with no statistically significant difference between the two groups (χ(2)=1.229, P=0.225). After successful resuscitation, the AST, ALT increased in both group but less in M group (all P<0.05). After successful resuscitation, the concentrations of TFN-α and IL-6 in the blood increased in both groups, reached the peak at 4h, and then decreased gradually. The concentrations of TFN-α in group M were lower than those in group C at 0.5, 2, 4 and 6 h after successful resuscitation (t=0.01, 0.01, 0.87, 0.86, all P<0.05). The concentrations of IL-6 in the group M were lower than those in group C at 0.5, 1, 2 and 4 h after successful resuscitation (t=0.23, 0.78, 0.11, 0.80, all P<0.05). Conclusions: After successful resuscitation, the release of inflammatory mediators, such as TNF-α and IL-6, and cell apoptosis may involve in liver ischemia reperfusion injury. After successful resuscitation, the liver undergoes ischemia-reperfusion injury, which may be related to the release of inflammatory mediators such as TNF-α and IL-6. Mild hypothermia therapy can prevent the release of TNF-α, IL-6 to reduce the degree of liver damage after resuscitation.

[Combined transperineal and transpubic urethroplasty for patients with complex male pelvic fracture urethral distraction defect].

OBJECTIVE: To evaluate the clinical effects and characteristics of combined transperineal and transpubic urethroplasty for patients with complex pelvic fracture urethral distraction defect (PFUDD). METHODS: We retrospectively reviewed the clinical data of 17 male patients with complex posterior PFUDD from January 2010 to December 2019. The complications included urethrorectal fistulas in 2 patients (11.8%), urethroperineal fistula in 1 patient (5.9%). Ten patients had undergone previous treatments: dilatation in 3 patients (17.6%), internal urethrotomy in 1 patient, failed urethroplasty in 6 patients (35.3%), of whom 2 patients had two times of failed urethroplasties. All the patients were performed with urethroplasty by combined transperineal and transpubic approach with removing the entire pubic bone followed by the anastomosis. RESULTS: The mean age of the patients included in this study was 35.5 (range: 21-62) years. The mean length of stricture was 5.5 (range: 4.5-7.0) cm, the mean follow-up was 27 (range: 7-110) months, the mean time of operation was 190 (range: 150-260) min, the mean evaluated blood loss was 460 (range: 200-1 200) mL. There were 5 patients who needed blood transfusion intraoperatively or postoperatively. Wound infection was seen in 4 out of 17 patients and thrombosis of lower extremities in 1 out of 17 patients. The last follow-up showed that the mean postoperative maximum urinary flow rate was 22.7 (range: 15.5-40.7) mL/s. After removing the catheter, one patient presented with decreased urinary flow and symptoms of urinary infection. Cystoscopy showed the recurrent anastomotic stricture, which was cured by internal urethrotomy. In our series, the success rate of the combined transperineal and transpubic urethroplasty was 94.1% (16/17). CONCLUSION: Combined transperineal and transpubic urtheroplasty can achieve a tension free anastomosis after removing the entire wedge of pubis in some patients with complex PFUDD. However, this procedure should be completed in a regional referral hospital due to the complexity of the operation and the high percentage of complications.

[Effect of lncRNA HOTAIR on the radiosensitivity of HCCLM3 cells].

Objective: To investigate the effect of down-regulating long non-coding RNA (lncRNA) and HOX transcript antisense RNA (HOTAIR) targeting miR-761 on the radiosensitivity of HCCLM3. Methods: The expression of HOTAIR in liver cancer cells HuH-7, SNU-449, HCCLM3 and normal liver cells L-02 were measured by real-time quantitative PCR. HCCLM3 cells were divided into control, Sh-NC (transfected shRNA negative control), Sh-HOTAIR (transfected HOTAIR shRNA), RAD+Sh-NC (transfected shRNA negative control and irradiated with 8Gy dose), and RAD+Sh-HOTAIR (HOTAIR shRNA was transfected and irradiated with 8Gy dose) group. Apoptosis was detected by flow cytometry, Bcl-2 Associated X Protein (Bx-2), cleaved cysteine-containing Cleaved cysteinyl aspartate specific proteinase 3 (C-Caspase-3) protein expression. Sh-NC, Sh-HOTAIR cells were irradiated with 0, 2, 4, 6, 8 Gy, and plate-clone experiments were used to determine radiosensitivity. Bioinformatics software predicted that miR-761 might be a target gene of HOTAIR, and the luciferase reporter system identified the targeting relationship. The miR-761 inhibitor, HOTAIR shRNA and inhibitor negative control, and HOTAIR shRNA were co-transfected into HCCLM3 cells, respectively. Cell apoptosis and Bax and C-cysteine-containing aspartate proteins were also measured using the above method, as well as the hydrolase-3 protein expression and cell survival fraction. Results: The expression levels of HOTAIR in liver cancer cells HuH-7, SNU-449, and HCCLM3 were higher than those in normal liver cells L-02 (1.85±0.12, 2.27±0.23, 2.68±0.15 vs 1.00±0.09, P<0.05). Compared with Sh-NC, the apoptosis rate of Sh-HOTAIR, RAD+Sh-NC cells and Bax, C-Caspase-3 protein levels are higher [Apoptotic rate: (13.47±1.32)%, (12.84±1.19)% vs (2.98±0.27)%; Bax protein: 0.74±0.08, 0.72±0.06 vs 0.42±0.06; C-Caspase-3 protein: 0.56±0.06, 0.54±0.08 vs 0.25±0.04, all P<0.05]. Compared with Sh-HOTAIR and RAD+Sh-NC, RAD+Sh-HOTAIR cell apoptosis rate and Bax, C-Caspase-3 protein levels are higher [apoptosis rate:(22.57±2.36)% vs (13.47±1.32)%, (12.84±1.19)%, Bax protein: 0.99±0.11 vs 0.74±0.08, 0.72±0.06, C-Caspase-3 protein: 1.03±0.12 vs 0.56±0.06, 0.54±0.08,all P<0.05]. Compared with Sh-NC, Sh-HOTAIR cells had lower survival scores and higher radiosensitivity (P<0.05). HOTAIR targets negative regulation of miR-761 expression. Compared with cells co-transfected with inhibitor negative control and HOTAIR shRNA, cells co-transfected with miR-761 inhibitor and HOTAIR shRNA had lower apoptosis rate after radiation treatment [(10.24±1.32)% vs (21.84±2.01))%], Bax (0.50±0.06 vs 1.01±0.10) and C-Caspase-3 protein (0.56±0.07 vs 1.05±0.14) had lower expression and higher cell survival scores (all P<0.05). Conclusion: Down-regulating lncRNA HOTAIR targets miR-761 to increase the radiosensitivity of HCCLM3.

[Clinical characteristics and prognosis of renal cell carcinoma patients with bone metastases].

Objective: To determine the status of bone metastasis (BM) and prognosis factors of patients with renal cell carcinoma (RCC) in our center. Methods: The clinical and medical records of RCC patients with BM, who were admitted to the Department of Urology, Bone Oncology and Spine Surgery, Beijing Jishuitan Hospital from August 2009 to August 2017 were collected. The gender, age, time of BM, location of BM, numbers of BM, presence or absence of visceral metastasis, pathological types of BM were investigated. The patients were followed up regularly, and the survival curves were analyzed by Kaplan-Meier method. Cox proportional hazard regression model was used to estimate the prognostic factors. Results: A total of 51 RCC patients with bone metastasis were collected. The age of patients ranged from 38 to 76 (58.6±8.2) years old, including 39 males (76.5%) and 12 females (23.5%). The ratio of male to female was 3.25∶1. The patients were followed up for 8 to 109 months, with a median follow-up time of 30 months. The follow-up rate was 90.2%. Thirty-one (60.8%) patients died at the last follow-up, with a median overall survival (OS) time of 25 months. The median OS was 38 months and 20 months in the solitary BM group (26 cases, 51.0%) and BM ≥ 2 group (25 cases, 49.0%), respectively. The difference between the two groups was statistically significant (P=0.021). The median OS was 30 months, 69 months and 17 months in the axis BM group (22 cases, 43.1%), appendicular BM group (19 cases, 37.3%) and both the axis and appendicular BM group (10 cases, 19.6%), respectively. The difference between the groups was statistically significant (P=0.012). The median OS was 22 months and 38 months in the patients with (15 cases, 29.4%) and without (36 cases, 70.6%) visceral metastases groups, respectively. The difference between the two groups was statistically significant (P=0.007). Univariate and multivariate Cox regression analysis showed that the numbers of BM (HR=3.130, 95%CI: 1.502-6.520, P=0.035) and visceral metastasis (HR=4.699, 95%CI: 1.810-9.545, P=0.001) were independent prognostic factors for RCC with BM. Conclusions: Solitary BM, no visceral metastasis are good prognostic factors for RCC with BM. For these patients, radical resection of BM is feasible to improve survival rate.

LncRNA CDKN2B-AS1 promotes the progression of ovarian cancer by miR-143-3p/SMAD3 axis and predicts a poor prognosis.

Long noncoding RNAs (LncRNAs) show great potential as the therapeutic targets attributing to their implication in the progression of various human cancers, including ovarian cancer (OC). Here, we aimed to explore the biological function of lncRNA cyclin-dependent kinase inhibitor 2B antisense RNA 1 (CDKN2B-AS1) in OC and its mechanism of action. The abundances of CDKN2B-AS1, miR-143-3p, and SMAD3 mRNA were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Cell Counting Kit-8 (CCK8) was performed to analyze cell proliferation. Cell apoptosis was assessed by flow cytometry and western blot analyses. Transwell assay was utilized to analyze cell migration and invasion abilities. Tumor xenograft was performed to confirm the role of CDKN2B-AS1 in ovarian tumor growth in vivo. The protein level of SMAD3 was examined by western blot assay. The interaction between CDKN2B-AS1 and miR-143-3p, or miR-143-3p and SMAD3 was demonstrated by bioinformatic, luciferase reporter, qRT-PCR and western blot analyses. CDKN2B-AS1 was upregulated in OC and correlated with clinicopathologic features. The knockdown of CDKN2B-AS1 hampered the development of OC, as reflected by the suppression of cell proliferation, migration, and invasion, and the enhancement of cell apoptosis, whereas the effects could be rescued by the overexpression of SMAD3. The absence of CDKN2B-AS1 blocked tumor growth in vivo. CDKN2B-AS1 served as a molecular sponge for miR-143-3p, leading to the derepression of miR-143-3p target SMAD3, which eventually triggered the progression of OC. In conclusion, CDKN2B-AS1 promoted tumor growth, invasion, and migration of OC by regulation of miR-143-3p/SMAD3 axis, hinting that CDKN2B-AS1 might be a potential biomarker for OC diagnosis and treatment.

Rationale for the advancement of PI3K pathway inhibitors for personalized chordoma therapy.

PURPOSE: Chordomas are rare and serious tumors with few effective treatments outside of aggressive surgery and radiation. Targeted therapies may present a more effective option for a subset of patients with lesions possessing certain genetic biomarkers. METHODS: A small molecule inhibitor library was tested in patient-derived UM-Chor1 cells to identify targeted therapies with potential efficacy. Targeted exome sequencing of UM-Chor1 and UM-Chor2 cells was performed to investigate genetic aberrations in relevant pathways. Chordoma cell lines were treated with inhibitors of the phosphotidylinositol 3-kinase (PI3K), epidermal growth factor receptor (EGFR), and cyclin dependent kinase (CDK) pathways, and responses were determined using resazurin cell viability assays, Annexin V apoptosis assays, and western blotting. Pan-PI3K inhibitor BKM120 was also tested in five chordoma xenograft models. RESULTS: Unbiased small molecule profiling nominated PI3K-AKT-mTOR pathway inhibitors as a promising therapy in chordoma, and genetic analyses of UM-Chor1 and UM-Chor2 cell lines revealed aberrations in PTEN, EGFR, and CDKN2A. Treatment of UM-Chor1 and UM-Chor2 with targeted PI3K, EGFR, and CDK inhibitors inhibited growth and proliferation and induced apoptosis more robustly than imatinib, a currently used chordoma therapy. Furthermore, BKM120 significantly inhibited tumor growth in a subset of the xenograft models tested. CONCLUSION: Targeted therapies, especially those inhibiting PI3K, display promising effects in multiple chordoma cell line and xenograft models. Nevertheless, the limited effects of PI3K, EGFR, and CDK targeting agents in other models reveal the presence of resistance mechanisms, which motivates future research to both identify biomarkers of response and develop combination therapies.

Prenatal diagnosis of maternal partial trisomy 9p23p24.3 and 14q11.2q21.3 in a fetus: a case report.

OBJECTIVE: This study aimed to report a fetus with maternal partial trisomy 9p and 14q and the phenotype detected in ultrasound. METHODS: The chromosome rearrangements in the fetus were characterized by G-banding and chromosome microarray analysis based on single nucleotide polymorphism (SNP) array of cultured amniocytes and compared with the parents' karyotypes. RESULTS: The fetal abnormal karyotype was 47,XY,+der(14)(9;14)(p23;q22). The SNP array revealed a duplicate 11.8-Mb 9p23-p24.3 fragment and a duplicate 29.6-Mb 14q11.2-q21.3 fragment. The peripheral blood karyotype of the mother was 46,XX,t(9;14)(p23;q22), while the father's was normal at the level of 300~400 bands. A high-resolution karyotype analysis conformed the same abnormality of the mother at the level of 550~650 bands. These results indicated that the fetal chromosomal abnormality probably derived from the mother. The fetal nuchal translucency thickness was 3.5 mm, and the fetal heart was detected with around 1.0-mm ventricular defect by the ultrasound examination at 12-week gestation. The couple decided to terminate the pregnancy. They opted for in vitro fertilization and embryo transfer for the fourth pregnancy, which was successful. CONCLUSIONS: The SNP array combined with cytogenetic analysis was particularly effective in identifying abnormal chromosomal rearrangements. These methods combined with the existing database information and fetal ultrasonography might provide a comprehensive and efficient way for the prenatal assessment of fetal situations. Preimplantation genetic diagnosis might effectively assist those women with an adverse pregnancy history in their next pregnancy.