ObjectiveTo optimize the processing technology of honey bran-fried Rosae Laevigatae Fructus（h-RLF）， formulate relevant quality standards， and explore its improving effect and mechanism on mice with ulcerative colitis（UC） induced by dextran sodium sulfate（DSS）. MethodsTaking the content of polysaccharides and water-soluble extract as the indexes， L₉（3⁴） orthogonal test was used to optimize parameters of the amount of honey bran， frying time and frying temperature. The quality of 15 batches of h-RLF decoction pieces was evaluated according to the optimized process， and the inspection limit standard was preliminarily drawn up. Eighty SPF male Kunming mice were randomly divided into 8 groups， including the blank group， model group， mesalazine group（0.13 g·kg^-1）， RLF group（3.77 g·kg^-1）， bran-fried RLF group（3.77 g·kg^-1）， h-RLF low， medium and high dose groups（1.89， 3.77， 7.54 g·kg^-1）， with 10 mice in each group. The mice in the blank group were free to drink pure water， and the other groups were free to drink 3% DSS solution for 7 days to prepare UC mouse model. Each treatment group was given corresponding drugs by intragastric administration， and the blank and model groups were given equal volume of normal saline. The body weight of mice was recorded daily and the disease activity index（DAI） was calculated. After the administration， the colon tissues of mice were collected to observe the pathological changes by hematoxylin-eosin（HE） staining. The levels of tumor necrosis factor（TNF）-α， interleukin（IL）-1β， IL-6 and IL-10 in the colon of mice were detected by enzyme-linked immunosorbent assay（ELISA）. Western blot was used to detect the expression levels of phosphorylation nuclear transcription factor-κB p65（p-NF-κB p65）， Toll-like receptor 4（TLR4）， p-p38 mitogen-activated protein kinase（p-p38 MAPK）， p-extracellular signal-regulated kinase（p-ERK） and p-c-Jun N-terminal kinase（p-JNK） proteins in colon tissues. ResultsThe optimum processing technology of h-RLF was 20 g honey bran per 100 g RLF， and stir-frying at 200 ℃ for 8 min. The limit standard under the examination of h-RLF was preliminarily formulated as follows：the polysaccharide content should not be less than 25% based on anhydrous glucose（C₆H₁₂O₆）， the content of water-soluble extract should not be less than 38%， the moisture content should not be more than 12.0%， the total ash content should not be more than 5.0%， and the acid-insoluble ash content should not be more than 1.0%. The cluster heat map analysis showed that the quality of RLF from Huanggang， Hubei province was better. Animal experiments showed that compared with the blank group， the DAI score of the model group was significantly increased， the levels of TNF-α， IL-1β and IL-6 in the colon tissue were significantly increased， the IL-10 level was significantly decreased， the colonic mucosa was seriously damaged， accompanied by a large number of inflammatory cell infiltration， tissue congestion and a significant reduction in glands， and the expression levels of p-NF-κB p65， TLR4， p-p38 MAPK， p-ERK and p-JNK proteins were significantly increased（P<0.01）. Compared with the model group， each administration group could alleviate the symptoms of colonic ulcer， the structure of colonic crypt was basically intact， and the glands were arranged in an orderly manner. Among them， the high-dose group of h-RLF had a better effect， which could significantly reduce the DAI score and the levels of TNF-α， IL-1β and IL-6 in colon tissue（P<0.01）， and significantly increase the level of IL-10（P<0.01）， alleviate the colonic mucosal injury， and effectively inhibit the expression levels of p-NF-κB p65， TLR4， p-p38 MAPK， p-ERK and p-JNK proteins（P<0.01）. ConclusionThe key parameters of the processing technology of h-RLF are determined， and the optimized technology is stable and feasible. The established quality standard is simple and reliable， and can be used for the quality control. h-RLF can effectively alleviate DSS-induced UC， and its mechanism may be related to inhibiting the activation of NF-κB/TLR4/MAPK pathway.

ObjectiveTo investigate the pharmacodynamic characteristics and explore the molecular mechanism of Honghua oral liquid （HOL） in relieving neuropathic pain （NP）. MethodHealthy male SD rats were randomly assigned into sham group， model group， low-， medium-， high-dose （0.5， 1.0， 2.0 mL·kg^-1·d^-1， respectively） HOL groups， and a positive drug （pregabalin， 25 mg·kg^-1·d^-1） group， with 6 rats in each group. Spinal nerve ligation （SNL） of L5 was conducted in other groups except the sham group. Drug administration was performed 3 days after the SNL surgery for 2 consecutive weeks， and samples were collected after the end of the administration. During the treatment period， the mechanical pain threshold and cold pain threshold were determined to measure the pain-relieving effect of HOL. Transcriptome sequencing was performed on hippocampal tissue samples from the sham， model， and high-dose HOL groups， and differentially expressed genes between the sham group and the model group as well as the model group and HOL high-dose group were obtained. After pathway enrichment analysis， we selected the targets which were closely related to neuroinflammation for validation， and predicted the specific binding sites of the major active components in HOL with the targets through molecular docking. In addition， the serum levels of tumor necrosis factor-α （TNF-α） and interleukin-10 （IL-10） were determined by enzyme-linked immunosorbent assay （ELISA） to evaluate the effect of HOL on neuroinflammation in NP rats. ResultCompared with the sham group， SNL decreased the mechanical pain threshold and cold pain threshold （P<0.05）. Compared with the model group， HOL recovered the mechanical pain threshold and cold pain threshold （P<0.05）. The transcriptome data showed that 376 differentially expressed genes （DEGs） were identified between the model group and the sham group， including 124 upregulated genes and 252 downregulated genes， and 194 DEGs between the model group and the high-dose HOL group， including 33 upregulated genes and 161 downregulated genes. Among them， insulin-like growth factor 1（IGF1）， matrix metallopeptidase-2 （MMP-2）， matrix metallopeptidase-14 （MMP-14）， erb-B2 receptor tyrosine kinase 2 （ERBB2）， and integrin subunit alpha 5 （ITGA5） associated with NP were selected for further validation. The Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） results showed that compared with the sham group， the modeling up-gurelated the mRNA levels of the above five molecules in the hippocampus （P<0.01）. Compared with model group， HOL down-regulated the mRNA levels of these molecules （P<0.01）. The molecular docking results showed that the main active components of safflower， hydroxysafflor yellow A， kaempferol， and quercetin， formed stable hydrogen bonds with the amino acid residues of IGF1， MMP-2， MMP-14， ERBB2， and ITGA5. The enzyme-linked immunosorbent assay（ELISA） results showed that compared with those in the sham group， the serum levels of TNF-α and IL-10 were out of balance in the model rats （P<0.01）. Compared with the model group， HOL lowered the level of the pro-inflammatory cytokine TNF-α （P<0.01） and elevated that of the anti-inflammatory cytokine IL-10 （P<0.05）. ConclusionHOL exerts analgesic effect on SNL rats by inhibiting neuroinflammation.

Objective:To explore the consistency of peripheral whole blood and venous serum procalcitonin (PCT) levels, and the value of peripheral whole blood PCT in evaluating pediatric bacterial infection.Methods:This multicenter cross-sectional parallel control study was conducted in 11 children′s hospital. All the 1 898 patients older than 28 days admitted to these hospitals from March 2018 to February 2019 had their peripheral whole blood and venous serum PCT detected simultaneously with unified equipment, reagent and method. According to the venous serum PCT level, the patients were stratified to subgroups. Analysis of variance and chi-square test were used to compare the demographic characteristics among groups. And the correlation between the peripheral blood and venous serum PCT level was investigated by quantitative Pearson correlation analysis.The PCT resultes were also converted into ranked data to further test the consistency between the two sampling methods by Spearman′s rank correlation test. Furthermore, the ranked data were converted into binary data to evaluate the consistency and investigate the best cut-off of peripheral blood PCT level in predicting bacterial infection.Results:A total of 1 898 valid samples were included (1 098 males, 800 females),age 27.4(12.2,56.7) months. There was a good correlation between PCT values of peripheral whole blood and venous serum ( r=0.97 , P<0.01). The linear regression equation was PCT?venous serum=0.135+0.929×PCT peripheral whole blood. However, when stratified to 5 levels, PCT results showed diverse and unsatisfied consistency between the two sampling methods ( r=0.51-0.92, all P<0.01). But after PCT was converted to ordinal categorical variables, the stratified analysis showed that the coincidence rate of the measured values by the two sampling methods in each boundary area was 84.9%-97.1%. The dichotomous variables also showed a good consistency (coincidence rate 96.8%-99.3%, Youden index 0.82-0.89). According to the severity of disease, the serum PCT value was classified into 4 intervals（<0.5、0.5-<2.0、2.0-<10.0、≥10.0 μg/L）, and the peripheral blood PCT value also showed a good predictive value (AUC value was 0.991 2-0.997 9). The optimal cut points of peripheral whole blood PCT value 0.5、1.0、2.0、10.0 μg/L corresponding to venous serum PCT values were 0.395, 0.595, 1.175 and 3.545 μg/L, respectively. Conclusions:There is a good correlation between peripheral whole blood PCT value and the venous serum PCT value, which means that the peripheral whole blood PCT could facilitate the identification of infection and clinical severity. Besides, the sampling of peripheral whole blood is simple and easy to repeat.

Objective To investigate the role of miR-374-5p in regulating mesenchymal stem cells ( MSCs) derived from gastric tissues to promote the proliferation and migration of gastric cancer cells in vitro. Methods The gastric cancer-mesenchymal stem cells ( GC-MSC) and the matched paracancerous tissue of gastric cancer derived normal mesenchymal stem cells ( GCN-MSC) were isolated and cultured in vitro. Gastric cancer cells were treated with the culture supernatants of GCN-MSC and GC-MSC respectively in vitro. The proliferation ability of gastric cancer cells was detected by plate clone test and the migration ability was detected by scratch test. MiR-374-5p mimics were used to over-express miR-374-5p in GC-MSC and GCN-MSC and MiR-374-5p inhibitor was used to inhibit its expression level. The culture supernatant of MSCs after transfection was collected and gastric cancer cells were treated in vitro. The changes of proliferation and migration ability of gastric cancer cells were observed. Results The number of single cell colonies formed in GC-MSC culture supernatant group (112±7) was more than that of GCN-MSC group (48±6) and the distance between the cells (6.5±0.5 cm) was less than that of GCN-MSC group (15±1) cm, by which the proliferation and migration of gastric cancer cells were significantly promoted (t=12.02, P<0.01 and t=13.17, P<0.01,respectively). Following miR-374-5p overexpressing in the culture supernatant of GCN-MSC,the number of single cell colonies increased (115±12.1) and space between cells reduced (7.83±1.08 cm), by which the proliferation ( t=9.31, P<0.01) and migration ( t=4.88, P<0.01) of gastric cancer were also promoted. The miR-374-5p inhibitor reversed the GC-MSC-induced ability of proliferation and migration of gastric cancer ( t=5.47,P<0.01 and t=4.95,P<0.01) . In the inhibitor group the number of single cell colonies decreased (60±10.2) and the space between cells increased (12.17±1.47cm) following treating by the culture supernatant. Conclusion MiR-374-5p should be an important molecule that regulates MSC in tumoral microenvironment to promote the proliferation and migration of gastric cancer and may become a prospective target for diagnosis and treatment of gastric cancer.

Objective To investigate the correlation between serum procalcitonin (PCT) levels and infection sites,as well as between PCT and bacterial species in gram negative (G-) bacteria induced sepsis,so as to provide rationale for therapeutic strategy of using antibiotic in sepsis.Methods The data of patients with sepsis admitted in Emergency Department and ICU from January 2014 to June 2015 were retrospectively analyzed.The blood culture of G-bacteria and PCT detection were carried out simultaneously within 24 hours after admission.The clinical data was analyzed to find out the correlation between PCT levels and infection sites,as well as between PCT levels and pathogenic bacterial species.Results A total of 187 specimens (came from 162 patients) were enrolled in the study with a median age of 70 years old and a median sequential organ failure assessment (SOFA) score of 4.PCT levels were found to be associated with bacterial species.PCT level caused by Escherichia coli bacteremia infection was higher than that caused by Acinetobacter baumannii bacteremia and Burkholderia cepacia bacteremia infection (4.62 ng/mL vs.2.44 ng/mL;4.62 ng/mL vs.0.81 ng/mL;P ＜ 0.05).Receiver operating characteristic (ROC) analysis showed an area under the curve (AUC) for PCT was 0.61 to discriminate Escherichia coli infection from Acinetobacter baumannii infection and an AUC was 0.66 to discriminate Escherichia coli infection from Burkholderia cepacia infection.When the cutoff point of PCT was 30.32 ng/mL,it could predict Escherichia coli infection rather than Acinetobacter baumannii infection with 94.10％ specificity,90.00％ positive predictive value and positive likelihood ratio for 4.24.When the cutoff point of PCT was 8.01 ng/mL,it could predict Escherichia coli infection rather than Burkholderia cepacia infection with 85.70％ specificity,93.94％ positive predictive value,and positive likelihood ratio for 3.01.When PCT cutoff value reached 47.31 ng/mL,the specificity and positive predictive value were both 100.00％.PCT level caused by urinary tract infection was higher than that caused by pulmonary infection (11.58 ng/mL vs.2.07 ng/mL,P ＜ 0.05),and the AUC was 0.69.When the cutoff point of PCT was 32.11 ng/mL,it could predict Escherichia coli infection rather than Acinetobacter baumannii infection with 90.60％ specificity,86.18％ negative predictive value and positive likelihood ratio for 3.68.Conclusions PCT elevation in G-bacteria induced sepsis might be associated with infection sites and bacterial species.

Objective To investigate the clinical features, diagnosis, treatment and prognosis of primary testicular diffuse large B-cell lymphoma.Methods 8 patients with primary testicular diffuse large B-cell lymphoma were reviewed from April, 2012 to April, 2015.The mean age was 58 years old, ranging 43-68 years old.Color Doppler echocardiography examination showed that there were round or oval regular tumors in the scrotum, which the diameter ranged from 3.5 to 8.0 cm, mean 5.5 cm.There was no abnormal changes among abdomen and pelvic cavity in CT scan and tumor markers examination.The radical orchiectomy were performed in all patients.After opening the tunica vaginalis, a hard texture tumor could be found, which has the vague border line with normal tissue.All patients were diagnosed according to the combination of morpbologic and immunohistochemical examination after operating.Results All patients accepted operation successfully.The mean operation time was 34 minutes, ranging 25-40 minutes.8 cases were diagnosed as primary testicular diffuse large B-cell lymphoma after operating.Immunohistochemical analysis showed that CD20, BCL-6 were positive.CD3, CD10, CK were negative.All patients received adjuvant chemotherapy with RCHOP(cytoxan 750 mg/m2, adriamycin 50mg/m2, leurocnstine 1.4 mg/m2, prednisone 60 mg/m2 ,rituximab 375 mg/m2) regimen over 6 cycles, which was conducted once every three weeks, one week post-operatively.The follow up duration ranged from 6 to 36 months, mean 17 months.All patients survived at the end of this study with no sign of recurrence and metastasis.Conclusions The patients with primary testicular diffuse large B-cell lymphoma are rare.The radical orchiectomy is recommended.And RCHOP chemotherapy should be considered one week post-operatively.The short term outcome of the treatment is satisfactory.But the long term outcome should be further studied.

Objective To study the role of early intravenous nutrition given aggressively combined with early minimal feeding on very low birth weight infants (VLBWI),and to evaluate the clinical value of intestinal barrier protein and MicroRNA.Methods All of 62 cases of VLBWI admitted in NICU,the Maternal and Child Health Hospital of Nantong Affiliated to Nantong University from January 2006 to June 2014 were recruited.Sixty-two VLBWI were randomly divided into group A and group B.Thirty infants in group A were exposed to conventional intravenous nutrition.Thirty-two infants in group B were treated with early intravenous nutrition aggressively combined with early minimal feeding.The time of birth weight recovery,days with intravenous nutrition,hospital stay and complications were recorded.The liver and kidney functions,electrolytes,blood gas analysis were monitored.Enzyme-linked immunosorbent method was used to detect intestinal fatty acid binding protein (Ⅰ-FABP),an intestinal barrier protein in plasma.Infection related MicroRNA155 was detected with fluorescent quantitative polymerase chain reaction (RT-PCR).Results Group B was superior to group A in weight loss after birth [(13.70 ± 3.10) ％ vs (5.46 ± 2.64) ％,P ＜ 0.05],shorter recovery time of body weight [(12.20 ± 3.38) d vs (6.82 ± 3.20) d,P ＜ 0.05],fewer days with intravenous nutrition [(29.62 ± 4.16) d vs (20.80 ± 3.20) d,P ＜ 0.05] and shorter hospital stays [(44.60 ± 6.32) d vs (28.91 ± 4.36) d,P ＜ 0.05].Compared with group A,the infants in group B had less complications,including hyperbilirubinemia (31.2％ vs 56.7％),extrauterine growth retardation (34.3％ vs 73.3％),cholestasis (6.2％ vs 23.3％),feeding intolerance (15.6％ vs 53.3％) and necrotizing enterocolitis (0 vs 16.7％) (all P ＜ 0.05).Although Ⅰ-FABP had a higher plasma concentration in group A than that of group B [(9.083 ± 1.059) μg/L vs (7.563 ± 0.739) μg/L],the difference was not significant (t =1.190,P =0.076 4).However,the plasma levels of Ⅰ-FABP in infants with necrotizing enterocolitis were significantly higher than those of group B [(19.500 ± 3.510) μg/L vs (7.563 ±0.739) μg/L,t =5.231,P =0.035 0].The expression of MicroRNA155 in group A was markedly higher than that of group B (2-△△ct were 0.81 ± 0.12 and 0.24 ± 0.08,respectively,P ＜ 0.05).Conclusions Giving aggressive intravenous nutrition early combined with early minimal feeding was safety and effective to VLBWI,which was of benefit to their growth and development,reducing complications and shorting hospital stays.The detection of intestinal barrier protein Ⅰ-FABP and MicroRNA155 is useful for monitoring feeding complications of VLBWI.

Objective:To detect the expression of microRNA-218 (miR-218) in human acute lymphocyte leukemia (ALL) T lymphocytes ( CCRF-CEM) ,explore its effects on the biological features of CCRF-CEM cells and the expression of its target gene c-kit, so as to provide new insights for leukemia treatment.Methods: Using the quantitative real-time polymerase chain reaction ( qRT-PCR) ,we detected the expression of miR-218 in the normal peripheral blood T lymphocytes and CCRF-CEM cells.Forty-eight hours after the miR-218 mimic was transfected into the CCRF-CEM cells,the expression of miR-218 in the CCRF-CEM cells was detected by qRT-PCR.The effect of miR-218 on the CCRF-CEM cell viability was detected using MTT.The effect of miR-218 on the proliferation and apoptosis of CCRF-CEM cell was analyzed using flow cytometry.c-kit gene was identified to be a target gene of miR-218 by luciferase reporter enzyme system,and the effect of miR-218 on the expression of KIT protein in cells were determined using Western blot.Results:As shown by qRT-PCR,compared with that in the normal peripheral blood T lymphocytes,the expressions of miR-218 in ALL T lymphocytes cell lines were significantly decreased ( P<0.01 ) .Compared with the control group, the expression of miR-218 increase significantly in CCRF-CEM cells transfected with miR-218 mimic for 48 hours ( P<0.01).MTT showed that the cell viability decreased significantly after the over-expression of miR-218 in the CCRF-CEM cells ( P<0.05 ) .Flow cytometry showed that the S-phase fraction significantly declined after the over-expression of miR-218 ( P<0.01 ) , and meanwhile the apoptosis of cells also significantly increased (P<0.01).Luciferase reporter gene assay showed that,compared with the control group,the relative luciferase activity significantly declined in the miR-218 mimic transfection group (P<0.01).Compared with the control group,the expression of KIT protein in the CCRF-CEM cells transfected with miR-218 mimic for 48 hours significantly decreased ( P<0.01).Conclusion:The expression of miR-218 decreases in ALL T lymphocytes cell lines.MiR-218 can negatively regulate the expression of KIT protein,inhibit the proliferation and increase the apoptosis of CCRF-CEM cells.Treatment based on the enhanced expression of miR-218 may be a promising strategy for leukemia.

Objective:To investigate the expression and significance of B cell activating factor (BAFF) and a proliferation-inducing ligand ( APRIL) in children with acute lymphoblastic leukemia ( ALL).Methods:The mRNA and protein expressions in ALL.

Objective To evaluate the effect of Tirofiban on CRP levels in patients with acute myocardial infarction (AMI) after primary emergency percutaneous coronary intervention (PCI). Methods Eighty-four AMI patients admitted on emergency were randomly divided into two groups: (1) early-treated group (n=45), immediately receiving Tirofiban intravenously on admission and (2) late-treated group (n=39), receiving Tirofiban intravenously after coronary angiography was performed. TIMI grading before and after PCI in beth groups were compared, CRP levels before and three days after PCI were estimated. The major adverse cardiovascular events (MACEs) occurred during hospitalization and following-up period of three months were recorded. Results Before PCI, TIMI grade 3 forward flow rate in early-treated group was significantly higher than that in late-treated group, while no significant difference existed between two groups after PCI. Three days after PCI, CRP level in early-treated group was markedly lower than that in late-treated group. During hospitalization, the occurrence of MACEs in early-treated group was lower than that in late-treated group, while no marked difference was found between two groups during the following-up period of three months. Conclusion In treating AMI patients with primary PCI, Tirofiban should be used as early as possible, which is safe and effective for PCI and can also significantly improve forward blood flow in target vessels, decrease the ClIP level and reduce the occurrence of MACEs during hospitalization.