[Surgical treatment and prognosis analysis of hilar cholangiocarcinoma].

Objective: To investigate the surgical treatment effect and prognostic factors of hilar cholangiocarcinoma. Methods: This is an ambispective cohort study. From August 2005 to December 2022,data of 510 patients who diagnosed with hilar cholangiocarcinoma and underwent surgical resection at the Hepatobiliary Center of the First Affiliated Hospital of Nanjing Medical University were retrospectively collected. In the cohort,there were 324 males and 186 females,with an age of (M (IQR)) 63(13)years (range:25 to 85 years). The liver function at admission was Child-Pugh A (343 cases,67.3%) and Child-Pugh B (167 cases,32.7%). Three hundred and seventy-two(72.9%) patients had jaundice symptoms and the median total bilirubin was 126.3(197.6) µmol/L(range: 5.4 to 722.8 µmol/L) at admission. Two hundred and fourty-seven cases (48.4%) were treated with percutaneous transhepatic cholangial drainage or endoscopic nasobiliary drainage before operation. The median bilirubin level in the drainage group decreased from 186.4 µmol/L to 85.5 µmol/L before operation. Multivariate Logistic regression was used to identify the influencing factors for R0 resection,and Cox regression was used to construct multivariate prediction models for overall survival(OS) and disease-free survival(DFS). Results: Among 510 patients who underwent surgical resection,Bismuth-Corlett type Ⅲ-Ⅳ patients accounted for 71.8%,among which 86.1% (315/366) underwent hemi-hepatectomy,while 81.9% (118/144) underwent extrahepatic biliary duct resection alone in Bismuch-Corlett type Ⅰ-Ⅱ patients. The median OS time was 22.8 months, and the OS rates at 1-,3-,5-and 10-year were 72.2%,35.6%,24.8% and 11.0%,respectively. The median DFS time was 15.2 months,and the DFS rates was 66.0%,32.4%,20.9% and 11.0%,respectively. The R0 resection rate was 64.5% (329/510), and the OS rates of patients with R0 resection at 1-,3-,5-and 10-year were 82.5%, 48.6%, 34.4%, 15.2%,respectively. The morbidity of Clavien-Dindo grade Ⅲ-Ⅴ complications was 26.1%(133/510) and the 30-day mortality was 4.3% (22/510). Multivariate Logistic regression indicated that Bismuth-Corlett type Ⅰ-Ⅲ (P=0.009), hemi-hepatectomy and extended resection (P=0.001),T1 and T2 patients without vascular invasion (T2 vs. T1:OR=1.43 (0.61-3.35),P=0.413;T3 vs. T1:OR=2.57 (1.03-6.41), P=0.010;T4 vs. T1, OR=3.77 (1.37-10.38), P<0.01) were more likely to obtain R0 resection. Preoperative bilirubin,Child-Pugh grade,tumor size,surgical margin,T stage,N stage,nerve infiltration and Edmondson grade were independent prognostic factors for OS and DFS of hilar cholangiocarcinoma patients without distant metastasis. Conclusions: Radical surgical resection is necessary to prolong the long-term survival of hilar cholangiocarcinoma patients. Hemi-hepatectomy and extended resection,regional lymph node dissection and combined vascular resection if necessary,can improve R0 resection rate.

[Efficacy and safety of neoadjuvant immunotherapy for hepatocellular carcinoma].

Objective: To study the surgical safety and efficacy of preoperative neoadjuvant therapy with immune checkpoint inhibitors combined with anti-angiogenic drugs in patients with China liver cancer staging(CNLC)-Ⅱb and Ⅲa resectable hepatocellular carcinoma. Methods: The data of 129 patients with Ⅱb and Ⅲa hepatocellular carcinoma who underwent surgery at the First Affiliated Hospital of Nanjing Medical University from January 2018 to December 2020 were analyzed. All patients were divided into two groups: the neoadjuvant therapy group(n=14,13 males and 1 female,aged (55.4±12.6)years(range:34 to 75 years)) received immune combined targeted therapy before surgery,immune checkpoint inhibitor camrelizumab was administered intravenously at a dose of 200 mg each time,every 2 weeks for 3 cycles,anti-angiogenesis drug apatinib was taken orally and continuously with a dose of 250 mg for 3 weeks and the conventional surgery group(n=115,103 males and 12 females,aged (55.8±12.0)years(range:21 to 83 years)) did not receive antitumor systemic therapy before surgery. There were 3 patients with CNLC-Ⅱb,11 with CNLC-Ⅲa in the neoadjuvant group;28 patients with CNLC-Ⅱb,87 with CNLC-Ⅲa in the conventional group. Student's t test or rank-sum test was used to compare the differences between two groups for quantitative data, Fisher's exact probability method was used to compare the differences of proportions between two groups, and Log-rank test was used to compare survival differences between two groups. Results: The 1-year recurrence rate in the neoadjuvant group was 42.9%,and the 1-year recurrence rate in the conventional group was 64.0%,with a statistically significant difference between the two groups(χ²=3.850,P=0.050);The 1-year survival rate in the neoadjuvant group was 100% and that in the conventional group was 74.2%,with a statistically significant difference between the two groups(χ²=5.170,P=0.023). According to the stratified analysis of the number of tumors,for single tumor,the 1-year recurrence rate in the neoadjuvant group was 25.0%,and that in the conventional surgery group was 71.0%,and the difference between the two groups was statistically significant(χ²=5.280, P=0.022). For multiple tumors, the 1-year recurrence rate in the neoadjuvant group was 66.7%,and the 1-year recurrence rate in the conventional surgery group was 58.9%,with no significant difference between the two groups(χ²=0.110,P=0.736). The operative time,intraoperative blood loss,and postoperative hospital stay in the neoadjuvant group were similar to those in the conventional group,and their differences were not statistically significant. Conclusions: Immune checkpoint inhibitors combined with anti-angiogenic targeted drugs as a neoadjuvant therapy for resectable hepatocellular carcinoma can reduce the 1-year recurrence rate and improve the 1-year survival rate,especially for those with solitary tumor. Limited by the sample size of the neoadjuvant group,the safety of immune combined targeted therapy before surgery cannot be observed more comprehensively,and further studies will be explored.

[Analysis of factors related to one-year mortality in elderly patients following hip fracture surgery].

Objective: To analyze the related factors that affect the one-year mortality in elderly patients following hip fracture surgery. Methods: The clinical data of the elderly patients who needed a surgery for hip fracture in the Second Affiliated Hospital of Wenzhou Medical University from January 2017 to April 2018 were retrospectively analyzed. According to the inclusion criteria of the study, 489 cases were included. Then the information of patients including age, sex, Charlson comorbidity index, type of fracture, Braden score at admission, American Society of Anesthesiologists (ASA) score, length of hospital stay, type of anesthesia, whole blood cells analysis were collected. Univariate and multivariate Cox regression analyses were conducted to investigate the factors related to one-year mortality of patients. Results: After excluding 39 patients, 450 patients were finally included. Patients in this cohort study had a mortality rate of 3.33% (15/450) at 1 month, 7.78% (35/450) at half a year, and 10.89% (49/450) at 1 year after surgery. Univariate analysis showed that age, sex, ASA score, type of fracture, Charlson comorbidity index, Braden score at admission, type of analgesia, preoperative hemoglobin concentration, preoperative albumin concentration, postoperative delirium of high activity correlated with one-year mortality after surgery. Further, multivariate Cox regression analysis revealed that age>80 years old (HR=2.32, 95%CI: 1.11-4.85, P=0.025), Charlson comorbidity index ≥ 3 (HR=3.24, 95%CI:1.75-6.03, P<0.001), Braden score at admission ≤16 (HR=1.93, 95%CI:1.03-3.57, P=0.040) and postoperative delirium of high activity (HR=2.49, 95%CI:1.16-5.35, P=0.019) were risk factors for one-year mortality. Conclusions: The current study indicates that one-year mortality rate of elderly patients following hip fracture surgery is 10.89%. Meanwhile, age>80 years, Charlson comorbidity index ≥ 3, Braden score at admission ≤ 16, postoperative delirium of high activity are risk factors for one-year mortality.

[Comparison of extracorporeal membrane oxygenation applicated in critical patients with COVID-19 and novel influenza A (H1N1) virus pneumonia].

Objective: To compare the clinical characteristics and prognosis of critical patients with COVID-19 and novel influenza A (H1N1) virus pneumonia (influenza pneumonia) applied with extracorporeal membrane oxygenation (ECMO). Methods: A total of 24 patients with influenza pneumonia treated with ECMO in respiratory intensive-care unit (ICU) of Beijing Chaoyang Hospital from March 2016 to December 2019 and 12 patients with COVID-19 hospitalized from February 1, 2020 to March 31, 2020 in 5 government designated infectious hospitals of Beijing and Hebei Province that applied with ECMO were enrolled. The demographic data, clinical manifestations, and ECMO related information were described and analyzed and all numerical variables are described as M (P25, P75). Results: The age of COVID-19 patients was 77 (66, 79) years old, which was older than influenza pneumonia patients [46 (32, 62) years old], P<0.05; acute lung injury score and respiratory ECMO survival prediction (RESP) score before ECMO application were 3.3 (3.0, 3.5) and 1 (0, 2), respectively, which were lower than influenza pneumonia patients [3.8 (3.5, 4.0) and 4 (2, 6), respectively], all P values<0.05. Thrombotic complications, bleeding complications, and ventilator-associated pneumonia occurred in ECMO applied COVID-19 patients were 4, 10 and 5 cases, respectively, which were more than that among influenza pneumonia patients (1, 9, and 2 cases, respectively), all P values<0.05. The length of ICU stay of COVID-19 patients was 31 (28, 75) d, which was longer than that of influenza pneumonia patients [27 (18, 39) d], P<0.05. The cases with successful decannulation of ECMO among COVID-19 and influenza pneumonia patients were 6 and 14 cases, respectively and mortality during ICU stay were 8 cases and 11 cases, respectively, and the difference were not statistically significant (all P values>0.05). Conclusion: COVID-19 patients applied with ECMO have more ECMO-related complications and a longer stay in the ICU than patients with influenza pneumonia.

[Surgical treatment of primary liver cancer:a report of 10 966 cases].

Objective: To summarize the experience of surgical treatment of primary liver cancer. Methods: The clinical data of 10 966 surgically managed cases with primary liver cancer, from January 1986 to December 2019 at Hepatobiliary Center, the First Affiliated Hospital of Nanjing Medical University, were retrospectively analyzed. The life table method was used to calculate the survival rate and postoperative recurrence rate. Log-rank test was used to compare the survival process of different groups, and the Cox regression model was used for multivariate analysis. In addition, 2 884 cases of hepatocellular carcinoma(HCC) with more detailed follow-up data from 2009 to 2019 were selected for survival analysis. Among 2 549 patients treated with hepatectomy, there were 2 107 males and 442 females, with an age of (56.6±11.1) years (range: 20 to 86 years). Among 335 patients treated with liver transplantation, there were 292 males and 43 females, with an age of (51.0±9.7) years (range: 21 to 73 years). The outcomes of hepatectomy versus liver transplantation, anatomic versus non-anatomic hepatectomy were compared, respectively. Results: Of the 10 966 patients with primary liver cancer, 10 331 patients underwent hepatectomy and 635 patients underwent liver transplantation. Patients with liver resection were categorized into three groups: 1986－1995(712 cases), 1996－2008(3 988 cases), 2009‒2019(5 631 cases). The 5-year overall survival rate was 32.9% in the first group(1986－1995). The 5-year overall survival rate of resected primary liver cancer was 51.7% in the third group(2009-2019), among which the 5-year overal survival rates of hepatocellular carcinoma, intrahepatic cholangiocarcinoma and mixed liver cancer were 57.4%, 26.6% and 50.6%, respectively. Further analysis was performed on 2 549 HCC patients with primary hepatectomy. The 1-, 3-, 5-, and 10-year overall survival rates were 88.1%, 71.9%, 60.0%, and 41.0%, respectively, and the perioperative mortality rate was 1.0%. Two hundred and forty-seven HCC patients underwent primary liver transplantation, with 1-, 3-, 5-, and 10-year overall survival rates of 84.0%, 64.8%, 61.9%, and 57.6%, respectively. Eighty-eight HCC patients underwent salvage liver transplantation, with the 1-, 3-, 5-, and 10-year overall survival rates of 86.8%, 65.2%, 52.5%, and 52.5%, respectively. There was no significant difference in survival rates between the two groups with liver transplantation (P>0.05). Comparing the overall survival rates and recurrence rates of primary hepatectomy (2 549 cases) with primary liver transplantation (247 cases), the 1-, 3-, 5-, and 10-year overall survival rates in patients within Milan criteria treated with hepatectomy and transplantation were 96.3%, 87.1%, 76.9%, 54.7%, and 95.4%, 79.4%, 77.4%, 71.7%, respectively (P=0.754). The 1-, 3-, 5-year recurrence rates were 16.3%, 35.9%, 47.6% and 8.1%, 11.7%, 13.9%, respectively(P<0.01). The 1-, 3-, 5-, 10-year overall survival rates in patients with no large vessels invasion beyond the Milan criteria treated with liver resection and transplantation were 87.2%, 65.9%, 53.0%, 33.0% and 87.6%, 71.8%, 71.8%, 69.3%, respectively(P=0.003); the 1-, 3-, 5-year recurrence rate were 39.2%, 57.8%, 69.7% and 29.7%, 36.7%, 36.7%, respectively (P<0.01). The 1-, 3-, 5-, and 10-year overall survival rates in patients with large vessels invasion treated with liver resection and transplantation were 62.1%, 36.1%, 22.2%, 15.0% and 62.9%, 31.8%,19.9%, 0, respectively (P=0.387); the 1-, 3-, 5-year recurrence rates were 61.5%, 74.7%, 80.8% and 59.7%, 82.9%, 87.2%, respectively(P=0.909). Independent prognostic factors for both overall survival and recurrence-free survival rates of HCC patients treated with liver resection included gender, neoadjuvant therapy, symptoms, AST, intraoperative or postoperative blood transfusion, tumor number, tumor size, cirrhosis, macrovascular invasion, microvascular invasion, and pathological differentiation. Propensity score matching analysis of 443 pairs further showed that there was no significant difference in overall survival rate between anatomical liver resection and non-anatomical liver resection(P=0.895), but the recurrence rate of non-anatomical liver resection was higher than that of anatomical liver resection(P=0.035). Conclusions: In the past decade, the overall survival rate of HCC undergoing surgical treatment is significantly higher than before. For HCC patients with good liver function reservation, surgical resection can be performed first, and salvage liver transplantation can be performed after recurrence. The effect of salvage liver transplantation is comparable to that of primary liver transplantation. As for the choice of liver resection approaches, non-anatomical resection can reserve more liver tissue and can be selected as long as the negative margin is guaranteed.

[Comparison of ultrasound-guided serratus anterior plane block and erector spinae plane blockperioperatively in radical mastectomy].

Objective: To compare the perioperative effects of ultrasound-guided serratus anterior plane block (SAPB) and erector spinae plane block (ESPB) in radical mastectomy. Methods: One hundred and fifty patients,undergoing radical mastectomy from May 2016 to Jan 2019,the Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, were randomly divided into SAPB group, ESPB group and control group. Patients in SAPB group and ESPB group were received corresponding blocks before induction of general anesthesia. The control group was only received routine general anesthesia without any block. Patient-controlled intravenous analgesia (PCIA) was performed in all the patients postoperatively. The VAS score at rest or coughing and Ramsay score at 2, 4, 8, 12, 24, 48 h after operation were compared among the three groups. The intraoperative dosages of propofol and remifentanil,press times and sufentanil cumulative dosage of PCIA in 48 hours after operation, postoperative rehabilitation indicators and adverse effects were all compared. Results: In all the three groups,the VAS scores at rest and coughing increased first and then decreased 2 h to 48 h after operation. The VAS scores in SAPB group and ESPB group were lower than that in control group (P<0.05), whereas, no significant difference was observed between SAPB group and ESPB group (P>0.05). For Ramsay score, among the three groups, there were no significances of the main effects of group and time point, as well as interaction effect (all P>0.05). The intraoperative dosages of propofol and remifentanil in SAPB group and ESPB group were lower than those in control group (P<0.05), the press times and sufentanil cumulative dosage of PCIA after operation were also lower than those in control group (P<0.05). There was no significant difference in feeding time after operation among the three groups (P>0.05). The times of first anal exhaust, ambulation and hospitalization after operation in ESPB group and SAPB group were significantly shorter than those in control group (P<0.05). However, there was no significant difference between ESPB group and SAPB group in postoperative rehabilitation indicators mentioned above (P>0.05). The incidences of skin itching and nausea in ESPB and SAPB groups were lower than those in control group (P<0.05). There was no difference in the incidence of vomiting among the three groups (P>0.05). Conclusions: Both SAPB and ESPB can provide good and safe analgesia for radical mastectomy,with equivalent performances in analgesia and adverse effect.

[A clinical study of mental stress-induced myocardial ischemia in patients with stable coronary artery disease and depression/anxiety].

Objectives: Exploring the association between depression/anxiety and mental stress-induced myocardial ischemia (MSIMI) in patients with stable coronary artery disease (CAD). Methods: A total of 178 subjects was enrolled according to the inclusion and exclusion criterion with 88 men and 90 women at age of (54±12)years. The subjects were divided into four groups including CAD with depression/anxiety, CAD without depression/anxiety, depression/anxiety without CAD, and control group based on the state of coronary artery, the scores of Patient Health Questionnaire 9-item (PHQ-9) and Generalized Anxiety Disorder 7-item (GAD-7) . MSIMI was diagnosed by echocardiography. Data were analyzed by SPSS19.0. Results: The incidence of MSIMI in all CAD patients was 17.24%. Within each group, 35.00% patients were MSIMI in CAD with depression/anxiety, 2.13% were in CAD without depression/anxiety, 14.29% were in depression/anxiety without CAD, and 2.38% were in control group. The risks of MSIMI in depression/anxiety without CAD and with CAD groups were 6.83 (P>0.05) and 22.08 times (P<0.05) higher than that in control group, respectively. Logistic regression showed that a 1-point increment in the GAD-7 score, but not PHQ-9 score [OR=0.95, 95% CI (0.77-1.17), P=0.63], was associated with 1.22-fold increase in the likehood of MSIMI [95% CI (1.07-1.38), P=0.00]. Conclusions: The MSIMI rate is much higher in patients with CAD comorbid depression/anxiety compared with CAD without depression/anxiety. Anxiety, but not depression, is an independent risk factor of MSIMI in CAD patients.

The Prognostic Significance of Soluble Programmed Death Ligand 1 Expression in Cancers: A Systematic Review and Meta-analysis.

The differential expression of soluble programmed death ligand 1 (sPD-L1) has been found in some cancers; however, the correlation between sPD-L1 expression and prognosis value in tumour is still unclear. Here, we conducted a meta-analysis and systematic review to assess the prognostic value of sPD-L1 in patients with cancer. Eligible studies were searched for in the databases including PubMed, Web of Science, ScienceDirect and Wiley Online Library database. The pooled hazard ratios (HRs) with a 95% confidence interval (95%CI) were calculated to assess the prognostic significance of sPD-L1 in human cancer. Eight studies and 1102 patients with cancer were included in the final analysis, and the combined analysis indicated that a higher level of sPD-L1 was associated with worse overall survival (OS) (HR = 1.60, 95%CI: 1.21-1.99). Furthermore, statistical significance was also observed in subgroup analysis stratified by the cancer type (haematological neoplasms or non-haematological neoplasms), sample size (more or less than 100), cut-off value of sPD-L1 (more or less than 6.51 ng/ml) and ethnicity (Asian or European). The meta-analysis indicates that circulating sPD-L1 changes may serve as a useful biomarker for cancer prognosis, and higher level of sPD-L1 may also be associated with poor outcomes in patients with cancer.

mRNA and protein expression levels of four candidate genes for ear size in Erhualian and Large White pigs.

Porcine ear size is an important characteristic for distinguishing among pig breeds. In a previous genome-wide association study of porcine ear size, LEM domain-containing 3 (LEMD3), methionine sulfoxide reductase B3 (MSRB3), high mobility group AT-hook 2 (HMGA2), and Wnt inhibitory factor 1 (WIF1) were implicated as important candidate genes for ear size. This study investigated the expression levels of four candidate genes for ear size in Erhualian and Large White pigs. Ten Erhualian pigs with large ears and eight Large White pigs with small ears at 60 days of age were examined. The mRNA expression levels of the four candidate genes were quantified by real-time polymerase chain reaction. WIF1 mRNA expression was significantly higher in Large White than in Erhualian pigs (P < 0.05), whereas the expression levels of the other three genes were not significantly different between the two breeds. The protein expression levels of the four genes were analyzed using western blot. WIF1 protein expression was significantly higher in Large White than in Erhualian pigs (P < 0.01), whereas MSRB3 protein expression was significantly higher in Erhualian than in Large White pigs (P < 0.05). There were no significant differences between the two breeds in residual protein expression. These results suggest that WIF1 is the main causal gene for ear size in pigs.

[Expression and clinical significance of HMGA2 in renal carcinoma].

Objective: To detect the high mobility group A2 (HMGA2) expression in renal carcinoma, and to explore the relationship with clinicopathological features and its significance for prognosis. Methods: 50 renal carcinoma specimens, 50 corresponding adjacent normal kidney tissue samples, and 40 benign renal tumor specimens were used in this study. The expressions of HMGA2 mRNA and protein were detected by RT-PCR, Western blot and immunohistochemical assays, and its relationship with clinicopathological features and prognosis in the renal carcinoma patients was analyzed. Results: The RT-PCR results showed that the relative expression levels of HMGA2 mRNA in the renal carcinoma, benign renal tumor tissues, and adjacent normal renal tissues were 0.84±0.23, 0.19± 0.06 and 0.08±0.04, respectively, and the expression in renal carcinoma tissue was significantly higher than those of the other 2 groups (P<0.01). The Western blot results showed that the relative expression levels of HMGA2 protein in the renal carcinoma, benign renal tumor tissues, and adjacent normal renal tissues were 0.91±0.24, 0.12±0.04 and 0.03±0.01, respectively, and the expression in renal carcinoma tissue was significantly higher than those of the other 2 groups (P<0.01). Immunohistochemical results showed that the expression of HMGA2 protein exhibited brown and tan granular, which mainly distributed in the cell nuclei. Among the 50 cases of renal carcinoma, 34 cases exhibited positive expression, with a positive rate of 68.0%. Among the 40 cases of benign tumor tissues, 3 cases had positive expression, with a positive rate of 7.5%, while among the 50 cases of adjacent normal renal tissues, there was only 1 case exhibiting positive expression of HMGA2 protein, with a positive rate of 2.0%. The protein expression of HMGA2 was significantly higher in the renal carcinoma than in the benign tumors and normal renal tissues (P=0.004). There was no statistically significant difference in the association of HMGA2 protein expressions with age, sex, tumor size and histological type (P>0.05), while significant difference did exist in the association with different statuses of TNM staging and lymph node metastasis (P<0.05). The median time to progression (TTP) in 34 HMGA2 protein-positive patients was (22.36±1.48) months and that of 16 HMGA2 protein-negative patients was (34.55±1.87) months (P<0.05). Conclusions: HMGA2 plays an important role in the tumorigenesis and development of renal carcinoma, and may be used as an important predictor for estimating the prognosis of renal carcinoma. HMGA2 might become a new diagnostic and prognostic marker for renal carcinoma.

Molecular cloning, tissue expression pattern, and copy number variation of porcine SCUBE3.

The signal peptide CUB EGF-like domain-containing protein 3 (SCUBE3) gene is a member of SCUBE gene family and plays important roles in bone cell biology and the determination of limb bone length. In this study, the full-length transcript of porcine SCUBE3 was cloned using reverse transcription-polymerase chain reaction and rapid amplification of cDNA ends. The full-length sequence of porcine SCUBE3 cDNA was 4131 base pairs and included 21 exons. The SCUBE3 gene contained a 2895-base pair open reading frame that encoded a peptide of 965 amino acids. Comparison of the deduced amino acid sequences of porcine SCUBE3 with those of human, mouse, zebrafish, and rat showed 96, 95, 73, and 95% identities, respectively. Porcine SCUBE3 mRNA expression levels were highest in the backfat, bone marrow, and cartilage tissues. Copy number variation was detected in porcine SCUBE3 and validated by real-time quantitative polymerase chain reaction. Different copy number variations were present in randomly selected individuals and may, therefore, be a good marker for identifying phenotypic traits. Our findings provide a basis for further investigation of the functions and regulatory mechanisms of SCUBE3 in pigs.

Porcine MAP3K5 analysis: molecular cloning, characterization, tissue expression pattern, and copy number variations associated with residual feed intake.

Mitogen-activated protein kinase kinase kinase 5 (MAP3K5) is essential for apoptosis, proliferation, differentiation, and immune responses, and is a candidate marker for residual feed intake (RFI) in pig. We cloned the full-length cDNA sequence of porcine MAP3K5 by rapid-amplification of cDNA ends. The 5451-bp gene contains a 5'-untranslated region (UTR) (718 bp), a coding region (3738 bp), and a 3'-UTR (995 bp), and encodes a peptide of 1245 amino acids, which shares 97, 99, 97, 93, 91, and 84% sequence identity with cattle, sheep, human, mouse, chicken, and zebrafish MAP3K5, respectively. The deduced MAP3K5 protein sequence contains two conserved domains: a DUF4071 domain and a protein kinase domain. Phylogenetic analysis showed that porcine MAP3K5 forms a separate branch to vicugna and camel MAP3K5. Tissue expression analysis using real-time quantitative polymerase chain reaction (qRT-PCR) revealed that MAP3K5 was expressed in the heart, liver, spleen, lung, kidney, muscle, fat, pancrea, ileum, and stomach tissues. Copy number variation was detected for porcine MAP3K5 and validated by qRT-PCR. Furthermore, a significant increase in average copy number was detected in the low RFI group when compared to the high RFI group in a Duroc pig population. These results provide useful information regarding the influence of MAP3K5 on RFI in pigs.

Molecular characteristics of the MHC-DRA genes from yak (Bos grunniens) and Chinese yakow (Bos grunniens × Bos taurus).

Two full-length cDNAs (762 bp) of the DRA gene from yak and Chinese yakow were isolated and analysed to identify structural and functional variations. The sequences for DRA in yak (Bogr-DRA) and Chinese yakow (Bogr × BoLA-DRA) were essentially identical to those for cattle (99%) and buffalo (97%). Except for two substitutions in the amino acids comprising the domain for signal peptide (SP) in yak, the additional residues were highly conserved across the species investigated. Peptide-binding site (PBS) of Bogr-DRA and Bogr × BoLA-DRA was highly reserved in the α1 domain among all species investigated. The lack of mutation in Bogr-DRA is consistent with the conception that the gene is highly conserved among all mammalian species. The very high conservation of the DRA gene among ruminants, including yak, may be due to its recent evolutionary detachment.

Analysis of differentially expressed genes in various stages of Duchenne muscular dystrophy by using a network view.

Duchenne muscular dystrophy (DMD), which is caused by mutations in the X-linked dystrophin gene, is a severe and progressive neuromuscular disease with no available cure. By integrating 2 microarray datasets from the Gene Expression Omnibus, we identified differentially expressed genes in 2 stages of DMD and systematically explored their potential disease-related mechanisms using a network view. Twenty differentially expressed genes were detected in various stages of DMD. According to the network with dystrophin as its center, none of the 20 proteins interacts with dystrophin directly. IQ motif-containing GTPase-activating protein 1 was found in the 2nd-level neighbors with a degree of 21. Microtubule-associated protein tau, membrane metallo-endopeptidase, interleukin 13 receptor alpha 1, and multiple epidermal growth factor-like domains 6 were found in the 3rd-level neighbors. These identifications require further investigation, as this report is the first of possible associations between DMD and these proteins. Analysis of differentially expressed genes through this network view may provide important information for further exploration of underlying mechanisms of DMD.

Author Correction: The dual role of anti-viral therapy in the treatment of Coronavirus disease 2019.

Correction to: European Review for Medical and Pharmacological Sciences 2020; 24 (22): 11939-11944-DOI: 10.26355/eurrev_202011_23854-PMID: 33275267, published online 30 November, 2020. The authors state that "Figures 3 and 4 were used twice due to a careless mistake during the preparation of Figures". There are amendments to this paper.  The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/23854.

Diversity of nitrogenase (nifH) genes pool in soybean field soil after continuous and rotational cropping.

Diazotrophs diversity in soybean is a topic requiring thorough investigation since the previous researches have focused on only rice, forest, grass, water, etc. In this research, iron-only nitrogenase nifH gene was as genetic marker. PCR-RFLP was used to investigate the difference of diazotrophs community diversity in the soil from the continuous cropping (CC) (the 5-yr tilling of soybean) and the rotational cropping (RC) (soybean-corn) soils in the northeast of China. A total of 36 isolates were genetically characterized. Most of the isolates closely related to Azospirillum and Azotobacter. Eighty-six unique nifH gene sequences were obtained by cloning of the respective PCR products in two soil samples. It was found that the diversity of nifH genes in CC changed obviously compared with RC. Phylogenetic analysis indicated that most of the clones clustered together in a high homogeneity with some sequence retrieved from environmental representatives. The sequence diversity of nifH genes was high and the members of the Alphaproteobacteria were predominant in both samples. The experimental study also revealed the two non-proteobacterial diazotrophs, firmicutes and euryarchaeota. Through this study, it can be assumed that different tillage perhaps affected the nifH gene-containing population diversity.

[Establishment of a dexamethasone-induced congenital cleft palate model in New Zealand rabbits].

Objective: To develop a new congenital cleft palate model suitable for the evaluation of cleft palate surgery and other related treatments. Methods: Ten New Zealand female rabbits (aged 40 weeks, 4.5-5.0 kg) were selected. The next day after mating with male rabbits of the same strain was regarded as the day 1 of gestation (GD1). Ten pregnant rabbits were enrolled with intramuscular injection 1.0 mg dosage of dexamethasone once a day from GD13 to GD16. The caesarean section was performed to obtain the newborn rabbits on GD31 for each pregnant rabbit. Then the rates of the survival and cleft palate rabbits were calculated. The rabbits were divided into two groups according to the method of random number table (10 non-cleft palate rabbits as the control group and 10 cleft palate rabbits as the experimental group). The body weights and physiological behaviors of the rabbits were evaluated and recorded at the age of 1, 2 and 4 weeks respectively after being fed by using standardized gastric tube feeding method. At 4 weeks old, three rabbits in each group were randomly selected for the observation of local anatomy of different layers of the mouth and upper jaw. The anatomical results were photographed for comparative analysis. Results: In this experiment, 48 infants of 10 pregnant rabbits survived under the condition with a survival rate of 66% (48/73), among which the incidence of cleft palate was 60% (29/48). All the rabbits in the control group and the experimental group were able to survive for at least 1 month with stable weight gain. There was no significant difference in weight (P>0.05) and physiological appearance between the two groups. In cleft palate group, most of fetuses showed complete cleft palate with significant differences in the anatomical structure of the upper jaw compared with the control group including the changes in the morphology of the palatal mucosa, the terminal distribution of the soft palate muscles, and the dysplasia and absence of bone structures along the mid-maxillary line. Conclusions: In this study, it was the first time to successfully establish the dexamethasone-induced congenital cleft palate model in New Zealand rabbits for cleft surgical research.