Aims To study the changes of serum me-tabolites in systemic lupus erythematosus ( SLE ) mice ( MRL/lpr) by treatment of Qinghao-Biejia and to ex-plore the pathogenesis of SLE and mechanism of drug action. Methods The serum samples of control group, SLE model group and Qinghao-Biejia treatment group ( low and high dose ) were collected, the metabolic profile of samples was analyzed by high performance liquid chromatography-quadrupole-time of flight mass spectrometry system ( HPLC-Q-TOF/MS ) . Software of Mass Hunter and Mass Profiler Professional ( MPP ) were used to process the data. A supervised mode of partial least squares-discriminant analysis ( PLS-DA ) was applied to recognize the data pattern. Results There were obvious disorders of lipid metabolism in SLE model. Compared with control group, Qinghao-Biejia treatment group improved lipid metabolism, af-fected the thrombosis development of SLE; and Qing-hao-Biejia treatment group reduced the pathological damage by improving inflammatory acute phase of SLE in mice. Conclusion Qinghao-Biejia treatment plays a therapeutic role in repairing the imbalance by multidi-mensional metabolic pathways in SLE mice.

CD56 Antigen ; metabolism ; Diagnosis, Differential ; Female ; Humans ; Immunohistochemistry ; Killer Cells, Natural ; metabolism ; pathology ; Leukocyte Common Antigens ; metabolism ; Leukosialin ; metabolism ; Lymphoma, Extranodal NK-T-Cell ; metabolism ; pathology ; Lymphoma, Non-Hodgkin ; metabolism ; pathology ; Middle Aged ; Skin Neoplasms ; metabolism ; pathology

Objective Influenza A(H7N9) virus causes a relatively high mortality in humans and therefore it is of great sig-nificance to know its prevalence in China .This article aimed to study the genetic characteristics and evolution of the hemagglutinin (HA) gene of the influenza A(H7N9) virus prevailing in China between 2013 and 2017. Methods We downloaded the HA se-quences of the influenza A ( H7N9) virus prevailing in China between 2013 and 2017 from The Global Initiative on Sharing All Influen-za Data and National Center for Biotechnology Information .Using the bioinformatics software , we analyzed the homology , molecular characteristics , phyletic evolution , and selective pressure of the HA gene. Results The homology of the HA gene of the influenza A ( H7N9) virus and the reference strain was decreasing each year from 2013 to 2017, 99.0%-99.9%in 2013, 98.7%-99.5%in 2014, 98.4%-99.6%in 2015, 76.8%-99.4%in 2016, and 69.9%-98.2%in 2017.Compared with the reference strain , the HA gene of the influ-enza A(H7N9) virus underwent variations in 21 antigenic sites.The variation of N285D was the highest (23%) in 2015 and that of R148K increased yearly,reaching 65%in 2016 and 78.5% in 2017. Phylogenetic analysis showed a concentrative distribution of the influenza A ( H7N9) virus strains on the phylogenetic tree in the same year from 2013 to 2017.Amino acid substitution of T 140A was observed in most of the influenza A ( H7N9) virus strains from Guang-dong in 2013, and the widest distribution of the virus strains was found in 2014.Positive selective pressure site 65 was obtained in the sequence of 2015 using the FEL and IFEL models, but not in the strains of 2016 or 2017. Conclusion Influenza A(H7N9) virus constantly undergoes variation , which has increased the difficulty in its prevention and control .More importance should be attached to observation of the virus and response to its adaptive mutations .

Objective:To study the dynamic changes of cellular immune function in peripheral blood of trauma patients and its role in the evaluation of traumatic complications.Methods:A prospective cohort study design was conducted. Patients with blunt trauma admitted to Chongqing Emergency Medical Center from November 2019 to January 2020 were consecutively enrolled. The peripheral blood samples were collected at 1, 3, 5, 7, and 14 days after injury. The expressions of CD64, CD274, and CD279 on the surface of neutrophils, lymphocytes, and monocytes as well as CD3 +, CD4 + and CD8 + T lymphocyte subsets were measured by flow cytometry. The trauma patients were divided into different groups according to the injury severity score (ISS) and sepsis within 28 days after injury, respectively. The dynamic changes of cellular immune function in different time points after injury and differences between different groups were compared. Furthermore, the correlation with acute physiology and chronic health evaluation Ⅱ (APACHEⅡ), sequential organ failure assessment (SOFA), and ISS were evaluated by Pearson correlation analysis. Results:A total of 42 patients with trauma were finally enrolled, containing 8 severe trauma patients with ISS greater than 25 scores, 17 patients with ISS between 16 and 25 scores, and 17 patients with ISS less than 16 scores. The sepsis morbidity rates were 14.3% (n = 6) within 28 days after injury. CD64 index and CD4 +T lymphocyte subsets were significantly increased at different time points after trauma (H = 15.464, P = 0.004; F = 2.491, P = 0.035). The CD64 index and positive rates of CD279 in neutrophils, lymphocytes, and monocytes were increased with the severity of injury at day 1 and day 3 after injury, respectively. At the first day after injury, CD64 index were 2.81±1.79, 1.77±0.92, 3.49±1.09; positive rate of CD279 in neutrophils were 1.40% (0.32%, 2.04%), 0.95% (0.44%, 2.70%), 12.73% (3.00%, 25.20%); positive rate of CD279 in lymphocytes were 3.77% (3.04%, 5.15%), 4.71% (4.08%, 6.32%), 8.01% (4.59%, 11.59%); positive rate of CD279 in monocytes were 0.57% (0.24%, 1.09%), 0.85% (0.22%, 1.25%), 6.74% (2.61%, 18.94%) from mild to severe injury groups, respectively. The CD64 index in severe injury group was significantly higher than that in moderate group, and the positive rates of CD279 in neutrophils, lymphocytes and monocytes of severe injury patients were higher than those in other two groups (all P < 0.05). At 3rd day after injury, compared to moderate group, severe injury patients had significantly higher CD64 index and positive rate of CD279 in lymphocytes [4.58±2.41 vs. 2.43±1.68, 7.35% (5.90%, 12.28%) vs. 4.63% (3.26%, 6.06%), both P < 0.05]. Compared with the non-sepsis patients, the sepsis patients had significantly higher CD64 index and positive rate of CD279 in monocytes at day 1 after injury [4.06±1.72 vs. 2.36±1.31, 3.29% (1.14%, 12.84%) vs. 0.67% (0.25%, 1.48%), both P < 0.05], and positive rate of CD279 in lymphocytes significantly higher at 3rd day after injury [8.73% (7.52%, 15.82%) vs. 4.67% (3.82%, 6.21%), P < 0.05]. In addition, correlation analysis showed that positive rate of CD279 in lymphocytes was positively correlated with SOFA and ISS, respectively (r values were 0.533 and 0.394, both P < 0.05), positive rate of CD279 in monocytes was positively correlated with APACHEⅡ, SOFA and ISS scores, respectively (r values were 0.579, 0.452 and 0.490, all P < 0.01), positive rate of CD279 in neutrophils was positively correlated with APACHEⅡ and ISS, respectively (r values were 0.358 and 0.388, both P < 0.05). Conclusions:CD64 index and CD279 expression in neutrophils, lymphocytes, and monocytes are significantly related to the severity and prognosis of trauma. Dynamic monitoring the cellular immune function may be helpful for assessing the prognosis of trauma patients.

Purpose To investigate the clinicopathological features, the diagnosis and differential diagnosis of spindle cell lipoma/pleomorphic lipoma (SCL/PL). Methods Eight cases of SCL/PL were studied by HE staining and immunohistochemistry. The literature was reviewed. Results Microscopically, the neoplasm consisted of spindle cells, mature adipocytes and thick rope-like collagen bundles in variable proportions with mucinous degeneration. The floret-like multinucleated giant cells were easy to be seen in the pleomorphic lipoma. Immunohistochemical analysis revealed that the tumor cells were positive for CD34 and vimentin. The Ki-67 was approximately 2％. The patients were followed up for 3 ～ 74 months, and no recurrence or metastasis occurred. Conclusion The SCL/PL is a special subtype of benign lipoma, surgery is considered the first choice, and outcome is favorable. The diagnosis and differential diagnosis of SCL/PL are important.

OBJECTIVETo investigate the relationship between the gene mutations of mannose binding protein(MBP) and the progression of hepatitis B.

METHODSThe MBP gene mutations in 52 patients with chronic hepatitis B and 62 patients with severe hepatitis B and 64 HBsAg-negative healthy controls were investigated. The mutations in MBP gene were analyzed by polymerase chain reaction (PCR) and direct DNA sequencing.

RESULTSA mutation of MBP gene codon 54 was found. The mutation frequency in the group of severe hepatitis B (35.5%, 22/62) was higher than those in the chronic hepatitis B group (15.4%, 8/52) and the HBsAg-negative healthy controls(14.1%, 9/64), respectively, and their difference was significant (chi-square was 7.79, P< 0.01; chi-square was 5.89,lzP<0.05). The difference between the chronic hepatitis B group and the HBsAg-negative healthy control group was not significant (P > 0.05).

CONCLUSIONThere is only mutation in codon 54 of the MBP gene in patients with hepatitis B infection in the area analyzed. Codon 54 mutation of MBP gene is not related to the persistence of hepatitis B, but it was associated with the progression of hepatitis B infection.

Adult ; Aged ; Codon ; genetics ; Female ; Hepatitis B ; genetics ; Humans ; Male ; Mannose-Binding Lectin ; genetics ; Middle Aged ; Mutation ; genetics ; Polymerase Chain Reaction ; Young Adult

Purpose To study the expression and the clinical significance of β3GnT8,MMP-2,PCNA in gastric cancer tissue. Methods The histological chips of paraffin specimens of gastric cancer were prepared,and immunohistochemical methods were used to detect the expression of β3GnT8,MMP-2 and PCNA in gastric cancer and adjacent tissue. Results The expression of β3GnT8 and MMP-2,PCNA in gastric cancer was higher than that in adjacent tissue (P = 0. 001) . The expression of β3GnT8 and MMP-2 was significantly positively correlated with the clinical stage (P = 0. 001) ,depth of invasion (P = 0. 011) and lymph node metastasis (P = 0. 003) of gastric cancer. The expression of β3GnT8 was significantly positively correlated with that of MMP-2 (r = 0. 703,P = 0. 001) and PCNA (r = 0. 231,P = 0. 024) . The overall survival time of the β3GnT8 positive expression group was significantly shorter than that of the negative expression group(χ2 = 3. 957,P = 0. 047) . Conclusion The expression of β3GnT8 is increased in gastric cancer. β3GnT8 can promote the invasion and metastasis of gastric cancer and is associated with poor prognosis in patients with gastric cancer.

OBJECTIVETo create a standard mini-swine model of chronic ischemic myocardium by endoscopy for the research of gene transfer and stem cell.

METHODSTwenty-three male China experimental minipigs were used, aged from 8 to 11 months with a mean of (9.3 +/- 1.8) months and weighed from 20 to 30 kg with a mean of (29.3 +/- 4.3) kg. The myocardial ischemia was established by gradual occlusion of the left circumflex coronary artery (LCX) with an Ameroid constrictor. The Ameroid constrictor was implanted around LCX by endoscopy. Selective coronary angiography, electrocardiogram and Echo-Doppler study were performed perioperatively to evaluate the degree of stenosis.

RESULTSChronic ischemic myocardial models were successfully generated in 20 of 23 swine by full-endoscopy. Ameroid constrictors were placed at the LCX accurately. Three swine died of anesthetic accident, cardiac arrhythmia at secondary coronary angiography, and pulmonary infection within 6 weeks after operation respectively. Operation time was 25 to 65 min with a mean of (46 +/- 9) min. The blood loss was 30 to 60 ml with a mean of (55 +/- 12) ml. Six weeks later, coronary angiography revealed the total occlusion and partial stenosis (> 85%) of the LCX occurred in 7 and 13 swine respectively. Cardiac systolic and diastolic dysfunction were found in all swine. The ejection fraction value was (65.0 +/- 6.3)% before operation and (41.0 +/- 9.3)% after operation (P = 0.008). The fractional shortening value was (36.2 +/- 4.3)% before operation and (34.2 +/- 2.3)% after operation (P = 0.027).

CONCLUSIONThe endoscopic surgery is a less invasive way to create a standard mini-swine model of chronic ischemic myocardium with effective results.

Animals ; Disease Models, Animal ; Feasibility Studies ; Male ; Myocardial Ischemia ; Swine ; Swine, Miniature ; Thoracoscopes

ObjectiveTo explore the effect of Chaishao Liujuntang on Hedgehog signaling pathway in rats with chronic atrophic gastritis （CAG） of liver depression and spleen deficiency. MethodWistar rats were randomized into normal group and modeling group. CAG with the liver depression and spleen deficiency syndrome was induced in rats in the modeling group with a compound method. After modeling， they were classified into the model group， vitacoenzyme group， Chaishao Liujuntang group， GDC-0449 （blocker） group， and Chaishao Liujuntang + GDC-0449 group. Normal group and model group were given （ig） normal saline. Vitacoenzyme and Chaishao Liujuntang group received （ig） corresponding drugs at 240 mg·kg^-1·d^-1 and 5.1 g·kg^-1·d^-1， respectively， and GDC-0449 group was treated （ip） with GDC-0449 at 50 mg·kg^-1·d^-1. For the Chaishao Liujuntang + GDC-0449 group， rats received GDC-0449 （ip） at 50 mg·kg^-1·d^-1 and Chaishao Liujuntang （ig） at 5.1 g·kg^-1·d^-1. The administration lasted 4 weeks. The pathological morphology of rat gastric mucosa was observed based on hematoxylin-eosin （HE） staining. mRNA and protein expression of sonic hedgehog （Shh）， 12th transmembrane receptor Patched1 （Ptch1）， and glioma-associated oncogene homolog 1 （Gli1） in gastric mucosa tissues was detected by real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） and Western blot. Content of serum interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α） was determined by enzyme-linked immunosorbent assay （ELISA）. ResultCompared with normal group， the model group demonstrated decrease in gland cells， glandular atrophy， large lumen volume， plasma cell infiltration， intestinal metaplasia， decrease in the mRNA and protein expression of Shh， Ptch1， and Gli1 in gastric mucosa （P<0.01）， and rise of serum IL-1β and TNF-α content （P<0.01）. Compared with model group， vitacoenzyme group and Chaishao Liujuntang group showed ordered cells， alleviation of gland atrophy， and no obvious inflammatory infiltration， and GDC-0499 group and Chaishao Liujuntang + GDC-0449 showed no significant improvement. Significant rise in the mRNA and protein expression of Shh， Ptch1， and Gli1 in gastric mucosa tissues of vitacoenzyme group and Chaishao Liujuntang group （P<0.01）， no significant difference in serum IL-1β content and significant decrease in TNF-α content in vitacoenzyme group （P<0.01）， significant reduction in content of serum IL-1β and TNF-α in Chaishao Liujuntang group （P<0.05， P<0.01） were observed compared with those in the model group. The mRNA and protein expression of Shh， Ptch1， and Gli1 in gastric mucosa and the content of serum IL-1β and TNF-α were insignificantly different between the GDC-0449 group and Chaishao Liujuntang + GDC-0449 group. ConclusionChaishao Liujuntang can effectively improve the pathological state of gastric mucosa in CAG rats with liver depression and spleen deficiency， which may be related to the activation of Hedgehog signaling pathway and the decrease of IL-1β and TNF-α content.

This paper aimed to investigate whether psoralen inhibits the differentiation and bone resorption by regulating CD4+T cell differentiation in RANKL-induced osteoclastogenesis in RAW264.7 cells, and elucidate its mechanism for osteoporosis. CD4+T cells were isolated from spleen cells of Balb/c mice by immunomagnetic separation method. The cells were divided into blank control group and psoralen group. The cells were cultured in 24-well plates and cultured for 3 days, and then they were collected for co-culture experiments after 4 days. Co-culture experiments were divided into RAW264.7 cell group, psoralen+RAW264.7 cell group, without psoralen treatment of CD4+T cells+RAW264.7 cell group, psoralen treatment of CD4+T cells+RAW264.7 cell group. After 5 days of co-culture, TRAP staining was used to detect the number of osteoclasts, and after 8 days of co-culture, bone resorption was evaluated by toluidine blue staining. The expressions of RORγt, Foxp3, IL-17, TNF-α, TGF-β and IL-10 in CD4+T cells and osteoclast differentiation-related genes MMP-9, TRAP and Cat-K were detected by Real-time polymerase chain reaction (RT-PCR); ELISA kit was used to detect IL-17, TNF-α, TGF-β and IL-10 and other cytokines levels. Our data confirmed that the psoralen significantly promoted the expression of Foxp3, TGF-β and IL-10 in CD4+T, and inhibited the expression of RORγt, IL-17 and TNF-α in CD4+T, the CD4+T cells without treatment by psoralen can significantly promote RANKL-induced differentiation of RAW264.7 to osteoclasts, and psoralen treatment of CD4+T can significantly inhibit RANKL-induced RAW264.7 osteoclast differentiation and bone resorption. Taken together, psoralen inhibits the differentiation and bone resorption of RAW264.7 into osteoclasts by promoting the development of CD4+ CD25+ Treg/Th17 balance in CD4+T cells to CD4+CD25+T.