Expressions of long non-coding RNA LINC00673 and ISG15 protein in pancreatic cancer and their clinical significances / 肿瘤研究与临床

Objective:To explore the expressions of long non-coding RNA LINC00673 and ISG15 protein in pancreatic cancer and their clinical significances.Methods:The clinical data of 57 patients diagnosed as pancreatic ductal carcinoma (PDAC) at the Affiliated Cancer Hospital of Xiangya Medical College of Central South University from January 2014 to December 2018 were retrospectively analyzed. The relative expressions of LINC00673 in pancreatic cancer tissues and paracancerous normal tissues (within 3 cm from the edge of cancer tissues) were examined by using quantificational reverse transcription-polymerase chain reaction (qRT-PCR). The ISG15 protein expressions in pancreatic cancer tissues and paracancerous normal tissues were examined by using immunohistochemistry. The difference in LINC00673 expression between ISG15 protein positive and negative patients was compared. The correlation between LINC00673 and ISG15 protein expressions in pancreatic cancer was analyzed by Spearman rank correlation analysis. Moreover, the correlations of LINC00673 and ISG15 protein expressions with clinical stage and pathological classification of pancreatic cancer patients were analyzed.Results:The positive expression of ISG15 protein in pancreatic cancer tissues was 40.4% (23/57), which was higher than that in paracancerous normal tissues [15.8% (9/57)] ( χ2 = 7.90, P = 0.004), and the relative expression of LINC00673 in pancreatic cancer tissues was 0.99±0.36, which was lower than that in paracancerous normal tissues (1.26±0.41) ( t = 4.80, P < 0.001). For 23 (40.4%) ISG15-positive patients and 34 (59.7%) ISG15-negative patients, the relative expression of LINC00673 was 0.77±0.46 and 0.45±0.27 ( P < 0.001). Spearman analysis showed that there was a correlation between LINC00673 and ISG15 protein expressions ( ρ = -0.429, P = 0.001). The relative expression of LINC00673 decreased in patients with low differentiated or undifferentiated tumor, vascular invasion and lymph node metastasis (all P < 0.05), but there was no correlation between LINC00673 expression and patients' age, tumor site, preoperative CA199 level, and TNM stage (all P > 0.05); ISG15 protein expression increased in patients with low differentiated or undifferentiated tumor, TNM stage Ⅲ-Ⅳ, vascular invasion and lymph node metastasis (all P < 0.05), but there was no correlation between ISG15 protein expression and patients' gender, age, tumor site, and preoperative CA199 level (all P > 0.05). Conclusions:The expression of LINC00673 in pancreatic cancer is related to vascular invasion, tumor differentiation degree and lymph node metastasis, and the expression of ISG15 in pancreatic cancer is related to vascular invasion, tumor differentiation degree, lymph node metastasis and TNM stage. The combined detection of LINC00673 and ISG15 protein could be a valuable prognostic indicator for pancreatic cancer. The therapies targeting LINC00673 and ISG15 protein signaling pathways are expected to be a potential option for immunotherapy of pancreatic cancer.

Clinical analysis on diagnosis and treatment of 116 patients with primary duodenal malignant tumor / 中国医师进修杂志

Objective:To explore the diagnosis, surgical methods and therapeutic effect of primary duodenal malignant tumor.Methods:The clinical data of 116 patients with primary duodenal malignant tumor from January 2010 to December 2018 were retrospectively analyzed.Results:Among 116 patients, adenocarcinoma was in 74 cases, interstitial tumor was in 25 cases, carcinoid was in 9 cases, the others was in 8 cases. Before operation, duodenoscopy was performed in 107 cases, and CT examination was performed in 76 cases. There were 57 cases of pancreaticoduodenectomy, 15 cases of duodenal segmental resection, 13 cases of subtotal gastrectomy and duodenal bulbar resection, 13 cases of duodenal partial resection, and 18 cases of palliative short circuit operation. The total incidence of postoperative complication was 31.9% (37/116), including pancreatic fistula in 8 cases (grade B 5 cases, grade C 3 cases), biliary fistula in 6 cases, abdominal infection in 5 cases, pulmonary infection in 4 cases, intestinal fistula in 3 cases, delayed gastric emptying in 3 cases, and hemorrhage in 8 cases. Four cases (3.4%) died during the perioperative period. Single factor Cox regression analysis result showed that the postoperative survival time was related to the tumor differentiation degree, operation method, tumor infiltration degree and lymphatic metastasis ( P<0.05 or <0.01); multi-factor Cox regression analysis results showed that the operation method, tumor infiltration degree and lymphatic metastasis were the independent risk factors for the postoperative survival time of patients with primary duodenal malignant tumor ( P<0.05). The patients were followed up until June 2021, and 9 cases were lost to follow-up. Kaplan-Meier survival curve analysis result showed that the postoperative overall 1-,3- and 5-year survival rates were 82.11%, 57.56% and 33.11%, respectively. Conclusions:Adenocarcinoma is the main primary malignant tumor of duodenum. Duodenoscopy and CT are the main examination methods. Radical resection is the most effective treatment for primary duodenal malignant tumor, and pancreaticoduodenectomy is the first choice. Surgical method, tumor infiltration degree and lymphatic metastasis are the independent risk factors affecting the prognosis of patients.

Interspecies Phylogenetic Analysis of / 生物医学与环境科学（英文）

Objective@#This study aims to investigate the infection of @*Method@#Infection of the definitive human host and intermediate fish host by @*Results@#In 2016-2020, the average population infection rate of Hunan was 1.38%, while in Tongdao County the rate was up to 26.90%, and the highest fish infection rate was detected in Qiyang County (99.44% in the dorsal fin of @*Conclusion@#The systematically study of

Effects of smoking on chronic obstructive pulmonary disease and respiratory symptoms / 中华疾病控制杂志

Objective To investigate the effects of smoking on chronic obstructive pulmonary disease(COPD) and respiratory symptoms. Methods A multi-stage, stratified cluster sampling strategy was used to select participants aged 40 or older in 5 surveillance points of Anhui Province. Questionnaires, body measurements and spirometry were used to collect data. Based on complex sampling design, Logistic regression model was conducted to analyze the effects of smoking on COPD and respiratory symptoms. Results The smokers who had smoked for ≥30 pack-years accounted for 13.9% (95% CI:10.3%-17.5%, P＜0.001) of the total population. And the smokers who had smoked for ≥40 years accounted for 8.5% (95% CI:6.7%-10.3%, P＜0.001) of the total population. On average, one smoker had smoked for 32.4 years (95% CI:31.2-33.5). Average daily cigarette consumption of daily smokers was 21.1 cigarettes (95% CI:19.6-22.7). As shown by multiple-variables Logistic regression analyses, the risk of COPD and respiratory symptoms increased with the increment of smoking pack-years and duration (all Ptrend <0.001). Conclusions Smoking was associated with COPD and respiratory symptoms. The risk of developing COPD and respiratory symptoms was greater with the increment of smoking pack-years and duration.

Effect of Heshi-Gejiugao on neuroendocrine immune network in women with perimenopausal syndrome / 国际中医中药杂志

Objective@#To study the mechanism of Heshi-Gejiugao on the nerve endocrine immune network in the treatment of perimenopausal syndrome.@*Methods@#A total of 100 patients with perimenopausal syndrome were randomly divided into treatment group and control group, 50 cases in each group. The control group was only given general treatment, while the treatment group was treated with Heshi-Gejiugao on the basis of general treatment for 30 days. The clinical efficacy, Kupperman score, nerve, endocrine and immune related indexes of the two groups before and after treatment were observed.@*Results@#The total effective rate was 96.0% (48/50) in the treatment group and 50.0% (25/50) in the control group. There was significant difference between the two groups (χ2=37.639, P<0.01). The Kupperman score (17.52 ± 2.73 vs. 24.22 ± 6.87, t=6.409) in the treatment group was significantly lower than that in the control group (P<0.01). After treatment, NE (1 878.08 ± 931.57 ng/m vs. 1 278.43 ± 866.32 ng/ml, t=3.331), DA (1 568.56 ± 597.15 ng/ml vs. 1 183.62 ± 798.72 ng/ml, t=2.729) in the treatment group were significantly higher than those in the control group (P<0.05); E2 (42.12 ± 9.77 pg/ml vs. 35.91 ± 12.55 pg/ml, t=2.761), FSH (62.70 ± 15.96 mIU/ml vs. 72.67 ± 30.18 mIU/ml, t=2.065), LH (33.88 ± 12.18 mIU/ml vs. 42.93 ± 9.83 mIU/ml, t=4.089) were significantly lower than those of the control group (P<0.05). CD3+ (1 087.34/μl ± 432.19/μl vs. 918.27/μl ± 199.72/μl, t=2.511), CD4+ (738.16/μl ± 326.75/μl vs. 588.43/μl ± 212.55/μl, t=2.716) and CD4/CD8 (1.87 ± 0.56 vs. 1.16 ± 0.55), t=6.483) were significantly higher than those of the control group (P<0.05); CD8+ (788.32/μl ± 214.56/μl vs. 976.37/μl ± 318.62/μl, t=3.462) was significantly lower than that of the control group (P<0.05).@*Conclusions@#Heshi-Gejiugao can reduce the symptoms and improve the quality of life by regulating the multi target and multi direction of the neuroendocrine immune network of perimenopausal patients.

Analysis of Risk Factors Affecting the Chemotherapy-related Infections in Patients with ALL / 中国实验血液学杂志

OBJECTIVE@#To analyze the risk factors affecting the chemotherapy-related infections in patients with acute lympho-blastic leukemia (ALL).@*METHODS@#The clinical data of 102 patients with ALL from January 2014 to December 2018 were collected and retrospectively studies. The risk factors of chemotherapy-related infections were analyzed by univa-riate and multivariate logistic regression.@*RESULTS@#A total of 386 courses of chemotherapy were completed, out of which the infection occurred in 201 course, with the infection rate of 52.07%, identified infection number was 215 case-times, including perianal infection of 13.95% (30/215), oral infection of 13.49% (29/215), blood flow infection of 1721% (37/215), lower respiratory tract infection of 37.21% (80/215), urinary infection of 3.26% (7/215), skin infection of 3.72% (8/215), digestive and intra abdominal infection of 9.30% (20/215), and other infections of 1.86 (4/215). Totally 88 strains of pathogenic bacteria were detected, including 29 Gram-positive bacteria (32.95%), 52 Gram-negative bacteria (59.09%) and 7 fungi (7.95%). Gram-positive bacteria mainly were Staphylococcus haemolyticus and Enterococcus faecium, susceptible to tegacycline, vancomycin and linezolid; Gram-negative bacteria mainly were Pseudomonas aeruginosa, Escherichia coli and Klebsiella pneumoniae, susceptible to tegacycline, amikacin, piperacillin/tazobactam and imipenem; Candida was the dominant fungus. Living in an ordinart ward, neutrophil defi-ciency for more than 7 days after chemotherapy and incomplete remission were independent risk factors of related infections during the induction chemotherapy in ALL inpatients, and hospitalization time also closely related with chemo-therapy-related infections in ALL inpatients (P<0.05). Neutrophil deficiency for more than 7 days after chemotherapy was an independent risk factor of chemotherapy-related infections in ALL inpatients in the consolidation chemotherapy (P<0.05).@*CONCLUSION@#Patients with ALL are prone to chemotherapeutic-related infections, and those who lack neutrophils for more than 7 days after chemotherapy and who do not reach complete remission are more prone to infection. Living in laminar flow ward and reducing hospitalization stay can help reduce the incidence of infection.

Effect of Modified Huanglian Jiedu Decoction Purgation Combined Electroacupuncture in Intervening Gastrointestinal Dysfunction of Critically Ill Patients Undergoing Abdominal Surgery / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effect of purgation and detoxification therapy on gastrointestinal dysfunction of critically ill patients undergoing abdominal surgery.</p><p><b>METHODS</b>Totally 56 inpatients with severe gastrointestinal dysfunction after abdominal surgery at ICU of Guangdong Provincial Hospital of Traditional Chinese Medicine were assigned to the treatment group and the control group, 28 in each group. All patients received routine Western medical treatment. Patients in the treatment group additionally took Modified Huanglian Jiedu Decoction (MHJD) and received electroacupuncture (EA) for 7 days. The first exhaust time, defecation time, scores for gastrointestinal dysfunction, mechanical ventilation time, ICU hospitalization time, and 28-day fatality rate were observed. Furthermore, serum levels of diamine oxidase (DAO) and D-lactic acid were detected at day 1, 3, and 7 after treatment.</p><p><b>RESULTS</b>The first exhaust time and the first defecation time in the treatment group were ahead of schedule, when compared with those of the control group (P <0. 05). Scores for gastrointestinal dysfunction, mechanical ventilation time, serum levels of DAO obviously decreased in the treatment group (P <0. 05). There was no statistical difference in serum levels of D-lactic acid, ICU stay time, the incidence of pulmonary infection, and 28-day mortality between the two groups (P >0. 05). Results of Logistic analysis showed that scores for gastrointestinal dysfunction were related with the incidence of pulmonary infection (P <0. 05).</p><p><b>CONCLUSION</b>MHJD combined EA could promote the recovery of gastrointestinal function in critically ill patients after abdominal surgery via improving intestinal barrier function, which was benefit for shortening mechanical ventilation time.</p>

Cloning, expression and purification of Schistosoma japonicum ribosomal protein S4 as well as the preliminary study of the diagnostic value of the recombinant protein / 中华预防医学杂志

<p><b>OBJECTIVE</b>To express and purify Schistosoma japonicum ribosomal protein S4(SjRPS4) in Escherichia coli, and assess its value in immunodiagnosis of Schistosomiasis japonica.</p><p><b>METHODS</b>Gene fragment of SjRPS4 was amplified by screening the cercaria cDNA library of Schistosoma japonicum. The target gene was cloned into the expressive vector pQE30 and transformed into E. coli M15. The recombinant protein expression was induced by isopropylthio-β-D-galactoside (IPTG). This fusion protein was purified by Ni(2+)-NTA chromatography and identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), Western blot and ELISA.</p><p><b>RESULTS</b>The plasmid pQE30/SjRPS4 was constructed successfully and expressed a SjRPS4 fusion protein in E. coli as showing a single special band on SDS-PAGE gel at Mr 30 × 10(3) position. It reached a purity of above 90% after purification. The Western blot result confirmed that the recombinant protein could specifically react with the serum samples from patients of schistosomiasis. Detecting the serum of Schistosomiasis japonica patients by ELISA, the sensitivity and specificity of the ELISA method were 90.91% (70/77) and 92.59% (25/27), the positive rate of recombinant protein expression was 67.30% (70/104). There was no cross-reaction with paragonimiasis patients' serum.</p><p><b>CONCLUSION</b>Protein SjRPS4 was successfully cloned and expressed, and it was confirmed that SjRPS4 antibodies were valuable in the diagnosis of Schistosomiasis japonica.</p>

Study on the immunoprotections of SjRPS4 and SjRPL7 DNA vaccines against Schistosoma japonicum infection in mice / 中国人兽共患病学报

The aim of this research was to study the immunoprotections of Schistosoma japonicum (S. japonicum) DNA vaccines SjRPS4 and SjRPL7 in mice. Fourty Kunming mice were randomly divided into 4 groups (A, B, C and D), and the pcDNA3.0/SjRPS4 and pcDNA3.0/SjRPL7 plasmid DNA vaccines were prepared for experiment. Mice in group A were intramuscularly injected with 100μL normal saline, whereas mice in group B were injected with 100 (g naked plasmid pcDNA3.0 into the quadriceps. Mice in groups C and D were injected with 100μg/100μL eukaryotic recombinant plasmids pcDNA3.0/SjRPS4 and pcDNA3.0/SjRPL7 into the hind leg muscles respectively. The initial injections were followed by two sets of boosters at 2 weeks intervals. In addition, levels of the specific antibodies were detected 2 weeks after the last immunization and all mice were percutaneously infected with 20( 1) S. japonicum cercariae on abdomen. Fourty-two days after the infection, all mice were killed to detect the worm reduction rate and the egg reduction rate. Significant differences of worm burden reduction rates, LEPG reduction rates, IEPG reduction rates and intrauterine eggs reduction rates were observed in both test group (group C and D), comparing with the control groups (group A and B). Results indicated that the DNA vaccines of pcDNA3.0/SjRPS4 and pcDNA3.0/SjRPL7 could induce strong protective immunity against S. japonicum in mice.

Detection of human papilloma virus 16/18 by the quantitative fluorescent PCR / 中华微生物学和免疫学杂志

Objective To study the relation between the HPV6/18 virus infection and the development of pathological changes of cervix. Methods The number of HPV16/18 DNA copies and the expression rate of HPV16/18 E7 mRNA in the pathological cervix were examined by the quantitative fluorescent PCR combined with pathological diagnosis and immunohistochemistry staining. Results The HPV16 infection rates in chronic cervicitis group were much lower (7.4%) than that in the cervical intraepithelial neoplasia (CIN) groups and the cervical cancer group (69.6% and 72.7%), respectively. Statistical analysis showed that the difference of HPV16 DNA copies was not significant between the chronic cervicitis group and CIN groups. In contrast to the above mentioned result, the number of HPV DNA copies between the CIN groups and the cervical cancer group was significantly different. The HPV16 E7 gene expression rates in CIN Ⅰ, Ⅱ, Ⅲ and cervical cancer groups were 0,37.5%,42.9%,63.6%, respectively. Conclusion Ins more common than that with HPV18. The number of HPV16 DNA copies in cervical cancer tissues is markedly higher than that in CIN Ⅱ, Ⅲ groups. The HPV16 E7 mRNA expression is significantly increased in the cervical cancer, and it is more closely correlated to this pathological changes. The quantitative fluorescent PCR can be used to reflect the activity of HPV, and it is a useful method for the screening examination of HPV and for the early diagnosis and treatment of cervical caner.

Cloning and expression of the succinate dehydrogenase iron-sulfur protein of Schistosoma japonicum Chinese strain in E. coli / 中南大学学报(医学版)

OBJECTIVE@#To clone the full-length gene encoding succinate dehydrogenase iron-sulfur protein of Schistosoma japonicum (SjSDISP) Chinese strain and express it in Escherichia coli.@*METHODS@#According to the published incomplete EST (BU804141) of SjSDISP and the sequence of multiclone sites of lambda gt11 vector, 2 pairs of primers were designed and synthesized. Then the 3' and 5'ends of the EST of the SjSDISP from adult Schistosoma japonicum cDNA library were amplified by anchored PCR. After sequencing, a full-length cDNA sequence of the SjSDISP was obtained, and then it was cloned into prokaryotic expression vector pGEX-4T-1. Identified by agarosed gel electrophoresis, endonucleases digestion and PCR, the resultant recombinant plasmid was used for the expression under the temperature-dependent condition and Western blot analysis.@*RESULTS@#A 1,071 bp sequence was obtained. Sequence analysis showed that the fragment contained a complete open reading frame (ORF), encoding 278 amino acid residues. This target fragment was cloned into the prokaryotic expression vector pGEX-4T-1, and expressed in Escherichia coli. SDS-PAGE revealed that the molecular weight of the expressed fusion recombinant product was 56 kD. Western blot showed that the recombinant protein was recognized by polyclonal rabbit antiserum immunized with Schistosoma japonicum adult worm antigen.@*CONCLUSION@#Cloning of the full-length gene encoding SjSDISP and its bacterial expression were successfully done.

Cloning,Expression and Immunization of The Hypoxanthine-guanine Phosphoribosyltransferase for Schistosoma japonicum Chinese Strain / 生物化学与生物物理进展

A 1 270 bp full-length cDNA fragment was obtained from the Schistosoma japonicum (Chinese strain) adult cDNA library after the 3′ and 5′ ends of the incomplete expression sequence tag (EST) of hypoxanthine-guanine phosphoribosyltransferase of Schistosoma japonicum (SjHGPRT) were amplified by the anchored PCR with 2 pairs of primer that were designed according to the published incomplete SjHGPRT EST and the sequence of multiclone sites of library ?gt11 vector. Sequence analysis indicated that this fragment, with an identity of 82% to hypoxanthine-guanine phosphoribosyltransferase of Schistosoma mansoni (SmHGPRT), contained a complete open reading frame(ORF). The deduced amino acid sequence showed 83% identity to that of SmHGPRT. This fragment was cloned into the prokaryotic expression vector pQE30, and subsequently sequenced and expressed in Escherichia coli. SDS-PAGE revealed that M of the recombinant protein was about 28 ku. Western-blot analysis showed that the recombinant protein was recognized by the polyclonal antisera from rabbits immunized with Schistosoma japonicum adult worm antigen. Mice vaccinated with recombinant protein revealed significant worm burden, liver eggs per gram (LEPG), fecal eggs per gram (FEPG) and intrauterine eggs of the female worms reduction percentage, compared with the controls. Taken together, the SjHGPRT full-length cDNA can be cloned and expressed in E.coli as a recombinant protein that elicited immunity against the challenge infection with Schistosoma japonicum, indicating its potential as a partial protection vaccine candidate.

Cloning, Expression and Immunization of The Hypoxanthine-guanine Phosphoribosyltransferase for Schistosoma japonicum Chinese Strain / 生物化学与生物物理进展

A 1 270 bp full-length cDNA fragment was obtained from the Schistosoma japonicum (Chinese strain) adult cDNA library after the '3' and 5' ends of the incomplete expression sequence tag (EST) of hypoxanthine-guanine phosphoribosyltransferase of Schistosoma japonicum (SjHGPRT) were amplified by the anchored PCR with 2 pairs of primer that were designed according to the published incomplete SjHGPRT EST and the sequence of multiclone sites of library λgt1 1 vector. Sequence analysis indicated that this fragment, with an identity of 82% to hypoxanthine-guanine phosphoribosyltransferase ofSchistosoma mansoni (SmHGPRT), contained a complete open reading frame(ORF). The deduced amino acid sequence showed 83% identity to that of SmHGPRT. This fragment was cloned into the prokaryotic expression vector pQE30, and subsequently sequenced and expressed in Escherichia coli. SDS-PAGE revealed that M of the recombinant protein was about 28 ku. Western-blot analysis showed that the recombinant protein was recognized by the polyclonal antisera from rabbits immunized with Schistosoma japonicum adult worm antigen. Mice vaccinated with recombinant protein revealed significant worm burden, liver eggs per gram (LEPG), fecal eggs per gram (FEPG) and intrauterine eggs of the female worms reduction percentage, compared with the controls. Taken together, the SjHGPRT full-length cDNA can be cloned and expressed in E. coli as a recombinant protein that elicited immunity against the challenge infection with Schistosoma japonicum, indicating its potential as a partia1 protection vaccine candidate.

Identification and analysis of effective compositions of Schistosoma japonicum 31-32 kDa proteins / 中国血吸虫病防治杂志

Objective To identify and analyze the effective compositions of Schistosoma japonicum 31-32 kDa proteins by using the techniques of proteomics.Methods The total proteins were prepared from 32-day adult worms of Schistosoma japonicum.After two-dimensional(2-D)gel electrophoresis,the distinct protein spots from 2-D gels were isolated and analyzed by MALDI-TOF-MS.Results A total of 13 protein spots,within the range of 31-32 kDa,were detected in the 2-D gels.Three of them had high homology with Actine-2 of S.mansoni,glycerol-3-phosphate dehydrogenase of S.japonicum and cathepsin B endopeptidase of S.mansoni.Conclusions The 31-32 kDa antigens contain 3 important antigens:actine-2,glycerol-3-phosphate dehydrogenase and cathepsin B endopeptidase,which have been demonstrated to have certain protective effect against S.japonicum.Our findings can facilitate the development of multi-epitope vaccine against S.japonicum.