ObjectiveTo reveal the molecular mechanism of Angong Niuhuangwan（AGNH） in improving traumatic brain injury（TBI） based on single cell sequencing. MethodSeventy-five male SD rats were randomly divided into the sham group， model group， piracetam group（3.6 g·kg^-1）， AGNH low- and high-dose groups（0.09， 0.27 g·kg^-1）， with 15 rats in each group. In addition to the sham group， the other 4 groups used the modified Feeney free-fall impact method to prepare TBI model， and the drugs were administered by gavage immediately after modeling， 24 hours later， the modified neurological deficit score（mNSS） was performed， and brain tissue was isolated to determine the degree of cerebral edema. Hematoxylin-eosin（HE） staining was used to observe the injury degree in the cortex， CA1 region and CA3 region of brain tissue. The expression levels of cyclooxygenase-2（COX-2）， interferon regulatory factor 1（IRF1）， Janus kinase 2（JAK2） and suppressor of cytokine signaling 3（SOCS3） were observed by immunofluorescence（IF） staining. The levels of interleukin（IL）-6， IL-18， IL-1β， IL-17A， tumor necrosis factor-α（TNF-α）， Caspase-1 and nucleotide binding oligomerization domain（NOD）-like receptor heat protein domain associated protein 3（NLRP3） inflammasome were determined by enzyme-linked immunosorbent assay（ELISA）. The regulation of AGNH on each cell population was analyzed by single cell sequencing， and differentially expressed genes were analyzed by Gene Ontology（GO） and Kyoto Encyclopedia of Genes and Genomes（KEGG）， which led to construct microglia differentially expressed gene network to search for the key targets， and validated by ELISA and IF. ResultCompared with the sham group， the mNSS and brain water content were significantly increased in the model group（P<0.01）. Compared with the model group， mNSS and brain water content in the low and high dose AGNH groups were decreased（P<0.05，P<0.01）. HE staining results showed that compared with the sham group， the cells in the cerebral cortex and hippocampus of rats in the model group were seriously lost， and the cells were arranged loosely（P<0.01）. Compared with the model group， AGNH could significantly increase the density of neurons in the CA1 and CA3 regions of the cerebral cortex and hippocampus， making the arrangement more compact， as well as improved cell morphology（P<0.05，P<0.01）. ELISA and IF staining showed that AGNH could reduce the levels of Caspase-1， IL-17A， TNF-α， NLRP3 and COX-2 in brain tissue of TBI rats（P<0.05， P<0.01）. A total of 13 cell subsets were identified by single cell sequencing， among which microglia played an important role in neuroimmunity. The results of GO enrichment analysis of differentially expressed genes in microglia showed that AGNH improved TBI in response to inflammation and TNF-α. KEGG enriched IL-17 signaling pathway， TNF signaling pathway， Toll-like receptor signaling pathway， etc. The results of network analysis showed that the key targets of AGNH in regulating TBI might be IL-6， IL-1β， JAK2， SOCS3， IRF1. IF and ELISA verification results showed that compared with the sham group， SOCS3 expression in microglia was decreased in the model group， and the expressions of IL-6， IL-1β， JAK2 and IRF1 were increased（P<0.01）. Compared with the model group， AGNH could increase the expression of SOCS3， decrease the expression of IL-6， IL-1β， JAK2， IRF1 （P<0.05， P<0.01）. ConclusionAGNH can reduce the degree of brain edema and brain injury， decrease the expression of inflammatory factors， and inhibit the expression of NLRP3 and its downstream Caspase-1 in TBI rats， which may act on the targets of IL-6， IL-1β， JAK2， IRF1 and SOCS3 in microglia.

Objective:To investigate the characteristics of multi-slice spiral CT images in lung cancer with cystic lesions and improve the accuracy of diagnosis.Methods:The clinical data of 27 cases of cystic lung cancer confirmed by pathological results of surgery or puncture who received treatment during 2012-2019 in Tongde Hospital of Zhejiang Province, China were collected. The clinical stages, pathological results and the characteristics of multi-slice spiral CT images were analyzed.Results:All 27 cases were diagnosed with peripheral lung cancer. Type VI peripheral lung cancer occurred in 12 (44.44%) cases, and adenocarcinoma in 17 (62.96%) cases. Stage Ia lung cancer occurred in 18 (66.67%) patients. Above 70% of CT images showed irregular cystic structure [70.37% (19/27)], polycystic structure [74.07% (20/27)], and vessels passing through the cysts [70.37% (19/27). Over 90% of CT images displayed unsmooth inner wall of the cysts [92.59% (25/27)] and fuzzy margin of the cysts [96.30% (26/27)].Conclusion:It is very difficult to diagnose lung cancer with cystic lesions. This kind of atypical lung cancer should be considered when the imaging features of peripheral lung cancer appear including thickened cystic wall, fuzzy cystic margin, vascular bundle sign and pleural indentation sign.

Objective:To investigate the current situation of kinesiophobia in patients with knee osteoarthritis and its influencing factors, in order to provide basis for clinical intervention.Methods:Totally 230 patients with knee osteoarthritis were selected from 2 tertiary hospital in Ji′nan by convenience sampling method, the self-designed general data questionnaire, Tampa Scale for Kinesiophobia, Numerical Rating Scale, Self-efficacy for Managing Chronic Disease, Hospital Anxiety and Depression Scale were used as research tools. Logistic regression analysis was used to analyze the influencing factors.Results:A total of 219 valid questionnaires were collected. Kinesiophobia occurred in 166 patients, with an incidence of 75.80% (166/219). Logistic regression analysis showed that gender, education level, chronic complications, pain and self-efficacy were the main influencing factors of kinesiophobia in patients with knee osteoarthritis ( P<0.05). Conclusion:The incidence of kinesiophobia in patients with knee osteoarthritis is relatively high, and medical staff should pay attention to the evaluation of kinesiophobia symptoms in this population, early identify the high-risk factors, provide timely health education, implement the appropriate intervention measures to reduce the occurrence of kinesiophobia.

Glycyrrhizae Radix et Rhizoma is one of the traditional herbal medicine used in China, study on the correlation between the cross-section color and HPLC fingerprints of them have important significance for promoting the development of traditional disciplines. Quantitative analysis of the color of sample cross section was carried out by color digital method, fingerprint analysis was carried out by HPLC, and the canonical correlation analysis was carried out between them. The results showed that there was a significant correlation between the color of Glycyrrhizae Radix et Rhizoma cross section and the information of HPLC fingerprinting. Results indicated that, The digitized indexes of color of cross section could reflect the result of fingerprint analysis to some extent.

Objective: To analyze the clinical characteristics of allergic reactions in patients with colorectal cancer after routine premed-ication with dexamethasone (5 mg) before chemotherapy with an oxaliplatin-containing regimen and to provide a reference for reduc-ing the risk of oxaliplatin allergy. Methods: Retrospective analysis of 242 patients with colorectal cancer who received chemotherapy with an oxaliplatin-containing regimen at the Changzhou Cancer Hospital of Soochow University from January 2014 to October 2017 was performed, and the incidence of allergic reactions was calculated. The associations between multiple factors and allergic reactions were then assessed using univariate analysis. The independent factors of allergic reactions were assessed using multivariate Logistic analysis, and the treatment outcome of oxaliplatin rechallenge in allergic patients was discussed. Results: A total of 242 patients were studied. They received routine premedication with dexamethasone (5 mg) before chemotherapy with an oxaliplatin-containing regi-men. Twelve (4.9%) patients had type I allergic reactions, with a median onset time of 6 (5.5-10.5) cycles and a median cumulative dose of 895 (716.5-1 075.0) mg. Multivariate analysis identified undergoing an oxaliplatin-free interval as an independent risk factor (P=0.04). Rechallenging patients with prophylactic agents was successful in 1 (33.3%) of 3 patients who completed their treatment. Conclusions: Routine low-dose dexamethasone premedication before oxaliplatin administration is safe and exhibited a lower inci-dence of hypersensitivity reactions than that reported in literature. An oxaliplatin-free interval may increase the risk of hypersensitivity reactions. Caution should be exercised while rechallenging allergic patients with oxaliplatin.

Integrins are cell adhesion receptors, which consists of several transmembrane glycoproteins. They are widely distributed on the cell surface and involved in signal transduction pathways. As a heterodimer, each integrin is composed of one α subunit and one β subunit. Integrins are mainly expressed on lepidopteran hemocytes and involved in cell immune response. The full-length cDNA sequence of BmIntegrin β2 was obtained by PCR and RACE, including 2 434 bp. BmIntegrin β2 was predicted to be a transmembrane protein. The BmIntegrin β2 expression profile was detected by qRT-PCR at L4D3 or L5D3 larval stage, and it was highly expressed in hemocyte and hematopoietic organ. Anti-BmIntegrin β2 polyclonal antibody was generated following prokaryotic expression, protein purification and animal immunization, which is highly specific and effective for recognizing BmIntegrin β2 protein through Western blotting. The results of plasmatocytes adhesion experiment showed that BmIntegrin β2 plays an important role on the adhesion and spreading of plasmatocytes to foreign surfaces. This study provides a foundation for further research of the biological function of BmIntegrin β2 gene.

Objective To observe the clinical value of application of low-dose scanning technology in dual source CT coronary angiography.Methods 200 cases who received coronary artery imaging were randomly selected as the study subjects.Patients were divided into the observation group and the control group using the random number method,100 cases in each group.All patients underwent dual source CT angiography.The observation group was treated by adaptive prospective ECG gated sequence,the control group was treated with retrospective ECG gated sequences.The image quality and radiation dose to bear were compared between the two groups.Results The various stages of image score proportion between the two groups had no significant difference(P＞0.05).The average score of reading image of the observation group was (1.52±0.33)points,which of the control group was (1.48±0.46)points,there was no difference between the two groups(t=0.857,P＞0.05).The CTDL,DLP and ED in the observation group were significantly lower than the control group[the observation group:CTDL(23.41±5.82),DLP(224.61±85.44),ED(3.01±1.12)mSv;the control group:CTDL(34.09±6.77),DLP(82.93±94.28),ED(5.33±2.01)mSv],and the differences were statistically significant(t=11.963,12.443,10.083,all P＜0.05)].Conclusion Prospective ECG gated sequence technology can effectively reduce the radiation dose of the patients under dual source CT coronary angiography,reduce probability of radiation damage,and the image quality is not changed obviously,which meets the needs of clinical diagnosis.

Objective: To optimize the extraction technology of Chinese medicinal materials in Yanyanling dispersible tablets.Methods: Orthogonal design was used to study the effects of three factors, including the ratio of liquid to materials, extraction time and extraction times on the extraction rate of Chinese medicinal materials and the comprehensive score of gallic acid content and total solid yield was used as the index.Results: The optimum extraction conditions were as follows: the ratio of materials to liquid was 1∶10 (g·ml-1), the extraction time was one hour, and the extraction times was three.Conclusion: Under the optimum conditions, the extraction rate of gallic acid in Chinese medicinal materials in Yanyanling dispersible tablets is 0.058%, and the total solid yield is 21.4%.The optimal process is stable and feasible, which can provide reference for the production of Yanyanling dispersible tablets.

Objective:To establish a liquid chromatography tandem mass spectrometry ( LC-MS/MS) method for the determination of 5-fluorouracil (5-Fu) in patient’s plasma and apply it in clinics patients validation. Methods:5-Fu was analyzed on an Agela Inno-val NH2 (2. 1 mm × 50 mm, 5 μm) column. Methanol:ultra pure water (2 ∶98) was used as the mobile phase with isocratic elution. The flow rate was 0. 3 ml ·min-1 and the column temperature was set at 40℃. The ion transitions with electrospray ionization negative model were m/z 128. 8→42. 1 and m/z 188. 6→42. 1 for 5-Fu and 5-bromouracil (the internal standard), respectively. The LC-MS/MS method was verified according to the guideline of quantitative analysis validation of biological samples ( Chinese Pharmacopoeia, 2015 edition, the fourth part) . Results:The calibration curve of 5-Fu was linear within the range of 10-1 000 ng · ml-1 . The lower limit of quantification was 10 ng · ml-1 . The precision, accuracy, matrix effect and stability within the linear range were all in line with the requirements of method validation. Conclusion:The LC-MS/MS method developed in the study for the determination of 5-Fu is simple, rapid, accurate and reproducible, which can be used for the plasma concentration detection of 5-Fu in patients.

Scavenger receptor class B is involved in various indispensable physiological processes, like the formation and inhibition of atherosclerosis or other cardiovascular diseases, innate immune defense and the removal of apoptotic cells. Here, we cloned BmSCRB8, a member of scavenger receptor class B in silkworm. We obtained the full-length cDNA sequence of BmSCRB8 by rapid amplification of cDNA ends (RACE), including 2 668 bp. The ORF of BmSCRB8 is 1 704 bp, encoding 567 amino acids. Online software prediction indicated that the molecular weight of BmSCRB8 is 63.87 kDa and the isoelectric point (pI) is 6.06. The space-time expression profile of BmSCRB8 was detected by reverse transcription PCR (RT-PCR), which implicated that BmSCRB8 is extensively expressed in each tissue and at each stage of blood. In addition, BmSCRB8 is highest expressed in fat body of silkworm, and is highly expressed in metamorphosis periods. Anti-BmSCRB8 polyclonal antibody was generated through prokaryotic expression, protein purification and mice immunization. Simultaneously, we constructed BmSCRB8 eukaryotic vector and then transfected embryonic cell line of silkworm. Immunofluorescence and overexpression showed that BmSCRB8 expressed specifically in membrane. Western blotting demonstrated that BmSCRB8 protein can be specifically recognized by anti-serum generated after mice immunization.