Objective:To explore the influence of central obesity on the risk of breast cancer and the possible role of dietary factors in its prevention.Methods:This study is a case-control study including a total of 212 participants, of whom 63 were with breast cancer, 71 were with breast nodules, and 80 were healthy controls. We used bioelectrical impedance analysis to measure body composition,and adopted the food frequency questionnaire to investigate dietary intake of participants.Results:The visceral adipose tissue ( OR=1.03, 95% CI: 1.003 to 1.077) and trunk fat mass ( OR=1.470, 95% CI: 1.104 to 2.184) were independently associated with the increased risk of breast cancer. Dietary patterns characterized by low dietary intake of beans and dairy products ( OR=1.300, 95% CI: 1.044 to 1.619) and high intake of cereals and red meat ( OR=2.254, 95% CI: 1.705 to 2.982) will increase the risk of breast cancer. Moreover, high meat intake ( β=0.268, 95% CI: 0.034 to 0.503) would advance the accumulation of visceral fat, while high bean intake ( β=-0.485, 95% CI: -0.865 to -0.104) would inhibit. Conclusions:Central obesity is an independent risk factor for breast cancer. Insufficient intake of beans and excessive intake of red meat are identified as factors that can exacerbate central obesity in breast cancer patients.

Reducing lactate accumulation has always been a goal of the mammalian cell biotechnology industry. When animal cells are cultured in vitro, the accumulation of lactate is mainly the combined result of two metabolic pathways. On one hand, glucose generates lactate under the function of lactate dehydrogenase A (LDHA); on the other hand, lactate can be oxidized to pyruvate by LDHB or LDHC and re-enter the TCA cycle. This study comprehensively evaluated the effects of LDH manipulation on the growth, metabolism and human adenovirus (HAdV) production of human embryonic kidney 293 (HEK-293) cells, providing a theoretical basis for engineering the lactate metabolism in mammalian cells. By knocking out ldha gene and overexpression of ldhb and ldhc genes, the metabolic efficiency of HEK-293 cells was effectively improved, and HAdV production was significantly increased. Compared with the control cell, LDH manipulation promoted cell growth, reduced the accumulation of lactate and ammonia, significantly enhanced the efficiency of substrate and energy metabolism of cells, and significantly increased the HAdV production capacity of HEK-293 cells. Among these LDH manipulation measures, ldhc gene overexpression performed the best, with the maximum cell density increased by about 38.7%. The yield of lactate to glucose and ammonia to glutamine decreased by 33.8% and 63.3%, respectively; and HAdV titer increased by at least 16 times. In addition, the ATP production rate, ATP/O₂ ratio, ATP/ADP ratio and NADH content of the modified cell lines were increased to varying degrees, and the energy metabolic efficiency was significantly improved.
Animals ; Humans ; L-Lactate Dehydrogenase/genetics* ; Lactic Acid ; Adenoviruses, Human ; Ammonia ; HEK293 Cells ; Glucose/metabolism* ; Adenosine Triphosphate/metabolism* ; Kidney/metabolism* ; Mammals/metabolism*

Objective:To explore the application value of whole-process ultrasound-guided percutaneous portal vein puncture islet transplantation.Methods:From October 2018 to May 2021, 16 diabetics underwent whole-process ultrasound-guided percutaneous portal vein puncture islet transplantation at First Affiliated Hospital of Sun Yat-sen University.The whole process was guided by ultrasound for completing percutaneous portal vein puncture catheterization, islet infusion monitoring, bleeding prevention and ablation hemostasis after bleeding.Results:Ten patients [8 males and 2 females with a mean age of(45.9±21.1)years]underwent 16 islet transplants, including one islet(5 cases), two islets(4 cases)and three islets(1 case). A single puncture was successfully performed without damage to other extrahepatic organs, persistent portal hypertension, portal vein embolism or infection.Bleeding at liver puncture site occurred in 3 cases and ultrasound radiofrequency ablation was performed for immediate hemostasis.Among them, postoperative blood glucose stabilized at 4~12 mmol/l post-operation.And 5 cases(31.3%)achieved insulin independence for>2 months and 10 cases(62.5%)lowered insulin dosage by>50% as compared with preoperative level.The level of fasting C-peptide recovered or was higher than normal in 10 cases(62.5%)and became obviously elevated in the remainders.In 11 cases(68.8%)of them, liver transaminase was briefly and mildly elevated post-operation, and no other complications were observed.Conclusions:The whole-process ultrasound-guided percutaneous portal vein islet transplantation is both safe and feseasible.It avoids the injury of transplanted kidney caused by contrast agent and radiological radiation to operator and patient.It is a method of islet transplantation worth a wider popularization.

Since December 2019, a novel coronavirus (2019-nCoV, SARS-CoV-2) pneumonia (COVID-19) outbreak has occurred in Wuhan, Hubei Province, and the epidemic situation has continued to spread. Such cases have also been found in other parts of the country. The spread of the novel coronavirus pneumonia epidemic has brought great challenges to the clinical practice of thoracic surgery. Outpatient clinics need to strengthen the differential diagnosis of ground glass opacity and pulmonary plaque shadows. During the epidemic, surgical indications are strictly controlled, and selective surgery is postponed. Patients planning to undergo a limited period of surgery should be quarantined for 2 weeks and have a nucleic acid test when necessary before surgery. For patients who are planning to undergo emergency surgery, nucleic acid testing should be carried out before surgery, and three-level protection should be performed during surgery. Patients who are planning to undergo emergency surgery in the epidemic area should be confirmed with or without novel coronavirus pneumonia before operation, and perform nucleic acid test if necessary. Surgical disinfection and isolation measures should be strictly carried out. Among postoperative patients, cases with new coronavirus infection were actively investigated. For the rescue of patients with novel coronavirus infection, attention needs to be paid to prevention and treatment and related complications, including mechanical ventilation-related pneumothorax or mediastinal emphysema, and injury after tracheal intubation.

Objective To analyze the biological characteristics of a mutant strain of Salmonella ty-phimurium SL1344 with sseK2-deletion (SL1344△sseK2) in order to provide reference for further study of safe and effective live vaccines. Methods The mutant strain SL1344△sseK2 with a deletion of 1047 bp in sseK2 gene was constructed through a two-step allelic exchange using recombinant suicide plasmid. Its com-plemented strain, SL1344C△sseK2, was also constructed. Biological and immunological characteristics of the mutant strain were detected. Results PCR, double-enzyme digestion and sequencing analysis showed that the mutant strain SL1344△sseK2 and the complemented strain SL1344C△sseK2 were successfully con-structed. The serotype of the mutant strain was 1,4,[5],12:i:1,2, identical to the parent strain SL1344. In addition, the mutant strain showed no significant change in biochemical characteristics or growth rate and was genetically stable in vitro. Compared with the parent strain SL1344, the virulence of SL1344△sseK2 was attenuated in BALB/ c mice. The median lethal dose of SL1344△sseK2 for 6-week-old BALB/ c mice was 3. 44×108 colony-forming units (CFU), which was 1620 times lower than that of SL1344. Oral immuniza-tion with SL1344△sseK2 protected 62. 5％ of the mice against challenge with wild Salmonella typhimurium strains on 17 d after vaccination. The levels of serum IgG antibody peaked on 14 d after immunization. No significant difference in biological characteristics was observed between the complemented and the parent strains, indicating that the mutant strain was basically complemented to the wild-type strain.Conclusions The mutant strain SL1344△sseK2 was constructed successfully and genetically stable with sig-nificantly attenuated virulence and good immunogenicity. This study suggested that sseK2 gene played an im-portant role in regulating the virulence of SL1344, which might provide reference for further study of its func-tion and for assessing its potential as a candidate live attenuated vaccine.

Objective To access the value of MRI in differential diagnosis between intrahepatic cholangiocarcinoma and atypical hepatic abscess. Methods Retrospectively collecting and analyzing the clinical and MRI imaging data of 19 patients with intrahepatic cholangiocarcinoma (ICC) and 17 patients with atypical hepatic abscess, confirmed by reexamination after anti‐inflammation therapy, surgery or puncture etiology, from June 2011 to July 2018 in Central Hospital of Lishui City.They were divided into ICC and abscess groups.All patients underwent routine liver plain MRI, DWI and contrast‐enhanced MR scan. The MRI features of the two groups (including morphology, boundary, cystic change and necrosis, pseudocapsule, hemorrhage, lipid composition, the signature of lesion in different phases of MRI and surrounding tissue) were studied. Fisher exact test and t test were used. Result This study showed that there was statistical difference between the two groups in the following aspects, the presence of cystic degeneration, the degree of annular enhancement in arterial phase, the homogeneous enhancement in portal venous phase and balanced phase and the central filling enhancement sign (P<0.05).The results showed that necrotic cystic lesion was more common in the abscess group (15/17 cases) than in the ICC group (0/19 cases);in the cases with annular enhancement in arterial phase,the degree of enhancement in the ICC group (13/16 cases) was higher than that in the abscess group (2/9 cases); the enhancement of the central parenchyma of lesion on out‐of‐phase images (1/19 cases) was slower in the ICC group than that in the abscess group (14/17 cases);and the ICC group was likely to present as central filling enhancement compared to the abscess group. Conclusion The presence of cystic lesions in DWI, the enhancement degree of marginal parenchyma, the enhancement speed of central parenchyma and the whole enhancement pattern are essential signs for differentiating intrahepatic cholangiocarcinoma and atypical hepatic abscess. 图1 女,53岁,右肝脓肿.病灶最大径4.0 cm,病灶中的小囊变区在DWI上呈高信号(↑) 图2 女,39岁,右肝脓肿.病灶最大径3.1 cm,病灶中的小囊变区在DWI上呈高信号(↑) 图3 女,66岁,右肝肝内胆管细胞癌(ICC).病灶最大径5.3 cm,横轴面T2WI病灶整体呈不均匀高信号,内见相对更高信号的富黏液区(↑) 图4 男,45岁,右肝ICC.病灶最大径5.8 cm,横轴面T2WI病灶整体呈高低混杂信号,内见散在片状低信号的凝固性坏死区(↑) 图5 与图1为同一患者.横轴面T2WI病灶实质部分呈均匀高信号,小囊变区呈明显高信号(↑) 图6 与图2为同一患者.横轴面T2WI病灶实质部分呈等信号,小囊变区呈明显高信号(↑)图7 与图3为同一患者.增强扫描动脉期横轴面示病灶边缘轻度不规则环形强化 图8 与图4为同一患者.增强扫描动脉期横轴面示病灶强化环有多处中断征象(↑) 图9,10 男,45岁,右肝ICC.病灶最大径5.8 cm,增强扫描动脉期横轴面(图9)示病灶边缘局部显著环形强化,局部强化环明显中断(↑).平衡期横轴面像(图10)示病灶中央有填充强化 图11 与图3,7为同一患者.平衡期横轴面示病灶内富黏液区出现轻微中央填充强化(↑) 图12 与图4、8为同一患者.平衡期横轴面像示病灶中央有填充强化 图13,14 男,59岁,右肝脓肿.病灶最大径4.3 cm,动脉期横轴面像(图13)示脓壁散在强化,周围见片状异常灌注.平衡期横轴面像(图14)示脓壁均匀强化,其内囊变区无强化 图15 与图2,6为同一患者.平衡期横轴面像示脓壁均匀强化呈相对等信号,其内囊变区无强化(↑) 图16 与图1,5为同一患者.平衡期横轴面像示脓壁均匀强化呈相对等信号,其内囊变区无强化(↑)

Objective To detect the expression and significance of serum miRNA-183 and TK1 in patients with colorectal cancer, and the mechanism thereof. Methods Fifty-two serum samples of colorectal cancer patients and paired health serum samples were collected. The expression of miRNA-183 was detected by real-time quantitative PCR, and TK1 was detected by Western blot enhanced chemiluminescence assay. The correlation between miRNA-183 and TK1 and their relations with the clinicpathologic characteristics were analyzed. Results The serum miRNA-183 expression was significantly higher in colorectal cancer group than that in normal control group (P<0.01). The expression of serum miRNA-183 was significantly higher in stage Ⅲ-Ⅳ group than that in stage Ⅰ-Ⅱ group (P < 0.01). There was more significant increase in serum miRNA-183 in lymphatic metastasis group than that without lymphatic metastasis group (P < 0.01). Receiver operating characteristic (ROC) curve showed that of there was a diagnostic value for serum miRNA-183 in colorectal cancer, with an optimal value of 1.15. The diagnostic sensitivity and specificity were 78.8%and 67.3%, and the positive predictive value and negative predictive value were 78.9%and 70.2%. The serum TK1 expression was also higher in colorectal cancer group compared with that of normal control group (P<0.01). And the TK1 expression was also higher in stageⅢ-Ⅳgroup than that in stageⅠ-Ⅱgroup (P<0.01). Furthermore, miRNA-183 expression was positively correlated with TK1 expression (rs=0.692, P<0.01). Conclusion The serum expression levels of miR-183 and TK1 may act as tumor markers for early colorectal cancer diagnosis, and also can be used to predict the malignant degree and prognosis of colorectal cancer.

Although there are 125 predicted DNase Ⅱ-like family genes in the Trichinella spiralis genome, plancitoxin-1-like (Ts-Pt) contains the HKD motif, a typical conserved region of DNase Ⅱ, in N- and C-terminal. It is generally believed that histidine is the active site in DNase Ⅱ. To study the nuclease activity of recombinant Ts-Pt with mutations in the active site from T. spiralis, different fragments of the mutated Ts-Pt genes were cloned using overlap PCR technique and inserted into the expressing vector pET-28a(+), and transformed into Escherichia coli Rosseta (DE3). The fusion proteins were purified by Ni-NTA affinity chromatography and SDS-PAGE. Nuclease activity of the recombinant proteins was detected by agarose gel electrophoresis and nuclease-zymography. The recombinant plasmids harboring the mutated Ts-Pt genes were constructed and expressed as inclusive body in a prokaryotic expression system. After renaturation in vitro, the recombinant proteins had no nuclease activity according to agarose gel electrophoresis. However, the expressed proteins as inclusive body displayed the ability to degrade DNA after renaturation in gel. And the nuclease activity was not affected after subjected to mutation of active site in N- and C-termini of Ts-Pt. These results provide the basis to study the relationship between DNase Ⅱ-like protein family and infection of T. spiralis.

Objective:To discuss the effects of recombinant IFN-α-IL-18 fusion protein on chicken lymphocyte histamine induced and the NF-κB p65 activation and nucleo-cytoplasmic transport.Methods: The healthy chickens blood was sterile adopted with anticoagulant,then separation of the chicken peripheral blood lymphocyte and divided into 10 groups:The yeast expression and purification protein IFN-α-IL-18,IL-18,IFN-α were added with 250 ng/ml,500 ng/ml and 1 000 ng/ml respectively while the control was only added RPMI1640 with 3 repetitions for each group.Then the histidine decarboxylase activity,histamine,IFN-γ,PI3K,MAPK and NF-κB p65 in cell nucleus were detected.Results: The recombinant IFN-α-IL-18 and IL-18 could significantly promote the activity of histidine decarboxylase (P<0.01),increase the contents of histamine (P<0.01),induce IFN-γ (P<0.01),improve the contents of PI3K (P<0.01) and the NF-κB p65 levels in nucleus (P<0.01),and the higher concentration of IFN-α had a similar effect to lymphocytes.The effects of IFN-α-IL-18,IL-18 and IFN-α on MAPK was acratia.Conclusion: IFN-α-IL-18 and IL-18 can stimulate chicken peripheral blood lymphocyte populations increased histamine contents significantly and promote the induction of IFN-γ.IFN-α-IL-18,IL-18 and IFN-α increase PI3K expression in lymphocyte associated with the NF-κB activation and NF-κB p65 nucleo-cytoplasmic transport.The study built foundation for the function of IFN-α-IL-18 and the exploration of the mechanism of controlling epidemic diseases.

Objective To study the sequence structure of Salmonella typhimurium Ssek3 gene and to express it at protein level in a prokaryotic expression system .Methods Sequence of Ssek3 gene was ob-tained from Salmonella typhimurium SL1344 strain.Bioinformatics methods were used for systematic analy-sis .A prokaryotic expression system for expressing Sse3k gene was constructed and the expressed protein was purified by Ni-NTA affinity chromatography .Results Sequence analysis showed that the Ssek3 gene of Sal-monella typhimurium was 1008 bp in length, encoding a protein of 335 amino acids and 72 amino acid resi-dues.The molecular weight, molecular formula and isoelectric point of Ssek3 protein was 37.89×103, C1700 H2629 N463 O497 S12 and 6.7, which indicated that it was a stable and hydrophilic protein .Ssek3 protein was a membrane protein without signal peptide or transmembrane region , containing five N-glycosylation sites , three O-glycosylation sites , 33 phosphorylation sites , 22 linear B-cell epitopes , 11 T-cell epitopes and 21 di-sulfide bonds.The secondary structure of Ssek3 protein contained 114 α-helices (Hh) (34.03％), 72 ex-tended chain (Ee) (21.49％), 30β-sheets (Tt) (8.96％) and 119 random coils (Cc) (35.52％).Re-sults of SDS-PAGE showed that the fusion protein Ssek 3 expressed in the prokaryotic expression system was a secretory protein with a molecular weight of about 40×103 .Conclusion The Ssek3 gene of Salmonella typh-imurium is successfully cloned , sequenced and expressed in this study , which will lay a foundation for fur-ther studying the role of Ssek3 protein in host cells during Salmonella typhimurium infection.