ObjectiveTo investigate the suppressive effect of 23-hydroxybetulinic acid （23-HBA）， a key constituent of Pulsatillae Radix， on the pulmonary inflammation-related carcinogenesis induced by the combined exposure of 4-（methylnitrosamino）-1-（3-pyridyl）-1-butanone （NNK） and lipopolysaccharide （LPS） in mice， alongside exploring its influence on immune cells and delving into the underlying mechanisms. MethodsA murine model of pulmonary inflammation-related carcinogenesis induced by NNK combined with LPS was established. Mice were randomly assigned into blank control， model， aspirin （10 mg·kg^-1）， and low-， medium-， and high-dose （3.75， 7.5， 15 mg·kg^-1， respectively） 23-HBA groups. The treatment lasted for 26 weeks， after which the spleen， lung， and peripheral blood samples were collected. Lung and spleen indices were calculated. Histopathological changes in the lung tissue were observed by hematoxylin-eosin staining. Immunohistochemistry was employed to assess the expression levels of thyroid transcription factor-1 （TTF-1）， neuron-specific enolase （NSE）， and proliferating cell nuclear antigen （Ki-67） in the lung tissue. High-throughput protein microarray was employed to measure the levels of interleukin-1β （IL-1β）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α） in the mouse serum. Flow cytometry was employed to evaluate the expression of macrophages， myeloid-derived suppressor cells （MDSCs）， and exhausted T lymphocytes in the lung and spleen tissue. Molecular docking was performed to predict the binding affinity of 23-HBA to Janus kinase 2 （JAK2）， Src homology 2 domain-containing phosphatase 2 （SHP2）， and suppressor of cytokine signaling 3 （SOCS3）. Western blot was performed to assess the protein levels of phosphorylated-signal transducer and activator of transcription 3 （p-STAT3）， p53， and SHP2 in the M1-activated macrophages and A549 lung adenocarcinoma cells treated with 23-HBA. ResultsCompared with the normal group， the lung and spleen indexes of the model group were increased to varying degrees （P<0.05， P<0.01）， the expression of TTF-1， NSE and Ki-67 protein was significantly increased （P<0.05， P<0.01）， and the serum levels of TNF-α， IL-1β and IL-6 were significantly increased （P<0.01）. The number of macrophages in the model group was significantly decreased （P<0.01）， and the number of exhausted T cells and MDSCs was significantly increased （P<0.05， P<0.01）. Compared with the model group， the spleen and thymus index of mice in each dose group of 23-HBA decreased significantly （P<0.05）， and the lung index of mice in the middle dose group of 23-HBA decreased significantly （P<0.05）. The high and middle dose groups of 23-HBA could improve the occurrence of inflammatory infiltration and malignant lesions in the lungs of mice induced by NNK combined with LPS in the model group. The expression of TTF-1 in the middle and high dose groups of 23-HBA was significantly lower than that in the model group （P<0.05， P<0.01）. The expression of NSE and Ki-67 protein in each dose group of 23-HBA was significantly lower than that in the model group （P<0.05， P<0.01）. The contents of IL-1β in the low and high dose groups of 23-HBA were significantly decreased （P<0.05）， and the contents of IL-6 and TNF-α in each dose of 23-HBA were significantly decreased （P<0.05， P<0.01）. The number of macrophages in the lung of the middle dose group of 23-HBA was significantly increased （P<0.05）， and the number of exhausted T cells and MDSCs expressing PD-1 in the lung was significantly decreased （P<0.05， P<0.01）. In addition， 23-HBA had strong molecular docking ability to SHP2， SOCS3 and JAK2 （≥7 kcal·mol^-1）， and significantly down-regulated the protein levels of p-STAT3， SHP2 and p53 in M1 macrophages and A549 lung adenocarcinoma （P<0.01）. Conclusion23-HBA holds promise as a potential therapeutic agent for mitigating pulmonary inflammation and inhibiting malignant transformation induced by the combination of LPS and NNK. It may exert effects by regulating immune cell responses， improving the tumor immune microenvironment， and regulating key signaling pathways.

ObjectiveTo investigate the protective effect and mechanism of Euphorbia helioscopia aqueous extract （EHE） on mice with chronic obstructive pulmonary disease （COPD） and its influence on precancerous lesion-associated proteins in lung tissues induced by cigarette smoke （CS）. MethodThe COPD model was induced by CS in 60 mice and the model mice were randomly divided into control group， model group， positive drug group （dexamethasone， 2 mg·kg^-1）， and low-， medium-， and high-dose EHE groups （1.875， 3.75， 7.5 g·kg^-1）. The high-performance liquid chromatography （HPLC） method was used to determine the related components in EHE. The changes in end-expiratory pause （EEP）， airway resistance （Penh）， expiratory flow at 50% vital capacity （EF50）， and other pulmonary function indexes were detected by the spirometer. The levels of inflammatory factors， such as interleukin （IL）-2， IL-5， IL-18， IL-17A， and IL-27 in bronchoalveolar lavage fluid （BALF） of mice were detected by high-throughput liquid protein chip technology. Hematoxylin-eosin （HE） staining was used to detect the pathological changes in lung tissues in mice. The content of malondialdehyde （MDA）， myeloperoxidase （MPO）， and glutathione peroxidase （GSH-Px） in lung tissues was determined by the colorimetric method. The mRNA relative expression of tumor necrosis factor-α （TNF-α）， transforming growth factor-β （TGF-β）， matrix metalloproteinase-2 （MMP-2）， matrix metalloproteinase-9 （MMP-9）， and matrix metalloproteinase-12 （MMP-12） was detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. Immunohistochemistry （IHC） was used to detect the expression of tumor protein （P53） and cell proliferation-associated antigen （Ki67） in lung tissues， and Western blot was used to detect the relative expression of tumor suppressor protein （P16）， DNA （cytosine-5）-methyltransferase 1 （DNMT1）， and fragile histidine triad （FHIT） in lung tissues. ResultThe results showed that the main compounds in EHE included phenols （gallic acid and protocatechuic acid） and flavonoids （such as hyperoside， rutin， myricetin， naringenin， quercetin， luteolin， kaempferol， and licorice chalcone A）， among which gallic acid and rutin were the highest in content. Compared with normal group， model group showed increased levels of EEP， EF50， and Penh （P<0.05）， and showed increased MDA and MPO levels （P<0.01） and decreased GSH-Px （P<0.01）， and the model group displayed increased levels of IL-2， IL-5， IL-18， IL-17A， IL-27， TNF-α， TGF-β， MMP-2， MMP-9， and MMP-12 （P<0.05）. And the model group exhibited up-regulated expression of P53， Ki67， and FHIT in lung tissues （P<0.01） and down-regulated expression of DNMT1 and P16 （P<0.01）. Compared with model group， the EHE groups showed decreased EEP and EF50 levels （P<0.05）. The pathological injury of lung tissues in mice of the model group was observed under HE staining， and the pathological injury of basal cell hyperplasia of lung tissues was gradually improved after treatment with EHE. The EHE groups showed reduced levels of MDA and MPO （P<0.01） and increased GSH-Px （P<0.01）. The EHE groups displayed decreased levels of IL-2， IL-5， IL-18， IL-17A， IL-27， TNF-α， TGF-β， MMP-2， MMP-9， and MMP-12 （P<0.05）. And the EHE groups showed down-regulated Ki67 and FHIT in lung tissues （P<0.05） and up-regulated expression of P53 and DNMT1 （P<0.05）. ConclusionEHE can protect mice from COPD and inhibit precancerous lesions， and the mechanism may be related to the inhibition of inflammation and oxidative stress response， regulation of protease and antiprotease imbalance， and regulation of epithelial cell growth.

Glioblastoma (GBM) is the most common and aggressive malignant brain tumor in adults and is poorly controlled. Previous studies have shown that both macrophages and angiogenesis play significant roles in GBM progression, and co-targeting of CSF1R and VEGFR is likely to be an effective strategy for GBM treatment. Therefore, this study developed a novel and selective inhibitor of CSF1R and VEGFR, SYHA1813, possessing potent antitumor activity against GBM. SYHA1813 inhibited VEGFR and CSF1R kinase activities with high potency and selectivity and thus blocked the cell viability of HUVECs and macrophages and exhibited anti-angiogenetic effects both in vitro and in vivo. SYHA1813 also displayed potent in vivo antitumor activity against GBM in immune-competent and immune-deficient mouse models, including temozolomide (TMZ) insensitive tumors. Notably, SYHA1813 could penetrate the blood-brain barrier (BBB) and prolong the survival time of mice bearing intracranial GBM xenografts. Moreover, SYHA1813 treatment resulted in a synergistic antitumor efficacy in combination with the PD-1 antibody. As a clinical proof of concept, SYHA1813 achieved confirmed responses in patients with recurrent GBM in an ongoing first-in-human phase I trial. The data of this study support the rationale for an ongoing phase I clinical study (ChiCTR2100045380).

Objective:To explore the effect of minimally invasivetransforaminal lumbar interbody fusion (MIS-TLIF) on lumbar multifidus muscle using MRI techniqueandits clinical significance.Methods:From September 2016 to July 2019, 23 patients who underwent MIS-TLIF surgery for unilateral symptomatic disc herniation in unilateral segments (L 3,4, L 4,5, L 5S 1) of Qingdao University Affiliated Hospital were studied. Their lumbar MR examination was performed 1 week before surgery, and 3 and 6 months after surgery. The axial section of multifidus muscle cross section area (AxCSA) was measured on the axial T2WI image of each intervertebral disc level before and after the operation. The ratio of long and short lines (RLS) was calculated, andthe ratio of axial section of muscle fat infiltration cross section area (FLSA) and AxCSAwasrecorded as FLSA/AxCSA. The changes of various indexes of multifidus muscle in the affected side and the healthy side of the lumbar spine before and after the operation were compared, and the effect of the MIS-TLIF procedure on the morphology of the multifidus muscle was observed. Magnetic resonance spectroscopy (MRS) measurements of the muscle cross-section of the affected side were performed before and 6 months after the operation. The integral value of intracellular lipid (IMCL) and extracellular lipid (EMCL) of 1H spectrum muscle cells was compared, while the degree of fat infiltration was measured. Results:Patients with single-segment lumbar disc herniation had larger AxCSAin the healthy side than the affected side before surgery in surgical level ( t=6.611, P<0.05), and the muscle AxCSAin the healthy side was larger than the affected side in non-surgical levels ( t=-6.682, P<0.05), both suggested preoperative muscle volume advantage in the healthy side; no difference in bilateral AxCSA at 3 months was found after surgeryin surgical levels ( t=0.197, P> 0.05)and non-surgical levels ( t=-1.631, P> 0.05), which suggested bilateral muscle volume equal advantageat short-term follow-up. The FLSA/AxCSA of affected segment before and after 3 months was 9.5%±3.8% and 8.7%±1.5%, and the difference was statistically significant ( t=3.163, P<0.05); the RLS of affected segment before and after 3 months was 3.3%±0.24% and 2.7%±0.83%, and the difference was statistically significant ( t=3.42, P<0.05). The medians of EMCL/IMCL before and after 6 months of MRS were 2.010 and 1.475, respectively, and EMCL decreased after 6 months ( Z=0.48, P<0.05). Conclusion:Patients with single-segment lumbar disc herniation have different morphology of bilateral multifidus muscle before surgery. MIS-TLIF has little effect on the multifidus muscle of the surgical side. MIS-TLIF significantly reduces extracellular lipid accumulation, promotes intracellular transfer, and increases intracellular fat metabolism. Its retention of muscle attachment points and limited fixation can also reshape compensatory muscle atrophy.

In the past ten years, the research and application of microbiome has continued to increase. The microbiome has gradually become the research focus in the fields of life science, environmental science, and medicine. Meanwhile, many countries and organizations around the world are launching their own microbiome projects and conducting a multi-faceted layout, striving to gain a strategic position in this promising field. In addition, whether it is scientific research or industrial applications, there has been a climax of research and a wave of investment and financing, accordingly, products and services related to the microbiome are constantly emerging. However, due to the rapid development of microbiome sequencing and analysis related technologies and methods, the research and application from various countries have not yet unified on the standards of technology, programs, and data. Domestic industry participants also have insufficient understanding of the microbiome. New methods, technologies, and theories have not yet been fully accepted and used. In addition, some of the existing standards and guidelines are too general with poor practicality. This not only causes obstacles in the integration of scientific research data and waste of resources, but also gives related companies unfair competition opportunity. More importantly, China still lacks national standards related to the microbiome, and the national microbiome project is still in the process of preparation. In this context, the experts and practitioners of the microbiome worked together and developed the consensus of experts. It can not only guide domestic scientific research and industrial institutions to regulate the production, learning and research of the microbiome, the application can also provide reference technical basis for the relevant national functional departments, protect the scale and standardized corporate company's interests, strengthen industry self-discipline, avoid unregulated enterprises from disrupting the market, and ultimately promote the benign development of microbiome-related industries.
China ; Consensus ; Humans ; Industry ; Microbiota

Objective:To prepare a dual targeting nanobubble both for HER2 target and cancer cell target, and compare the targeting ability with single targeting nanobubble either with HER2 targeting or cancer cell targeting ability in vitro.Methods:Nanobubbles(NBs) were prepared using the modified thin film hydration method and then connected with IR783 directly (NBs-IR783), HER2 antibody Affibody by avidin-biotin method (NBs-Affibody), or both of them (IR783-NBs-Affibody). The size distribution, stability, and biosafety of these contrast agents were observed. Flow cytometry and immunofluorescence were applied to compare the binding rate of these NBs against HER2 positive breast cancer cell in vitro.Results:The average size of NBs-Affibody, NBs-IR783 and IR783-NBs-Affibody was (538.4±95.8)nm, (551.8±114.8)nm, and (482.7±54.3)nm, respectively. The binding rate for HER2 positive cell was 26.6%, 97.6%, 84.5%, while for HER2 negative cell was 5.4%, 99.9% and 99.3%, respectively. The results of dual-labeled flow cytometry showed that the binding rate of IR783-NBs-Affibody mediated by IR783 to HER2 positive breast cancer cells were 79.5% and Affibody mediated HER2 targeting binding rate were 19.4%. However, NBs-IR783 only showed IR783-mediated tumor targeted binding without HER2 targeting ability with binding rate of 2.3%.Conclusions:The prepared IR783-NBs-Affibody has the dual targeting ability for HER2 positive breast cancer cell, which is superior to the single targeting NBs (NBs-Affibody and NBs-IR783). IR783-NBs-Affibody is optimal to meet the imaging requirements of tumor heterogeneity.

Objective To study the feasibility of laparoscopic pancreaticoduodenectomy ( LPD) in the treatment of pancreatic head cancer, and to analyze the short-term postoperative outcomes. Methods The clinical data of 57 patients with pancreatic head cancer who underwent laparoscopic pancreaticoduode-nectomy from April 2015 to November 2017 in the Second Hospital of Hebei Medical University were retro-spectively analyzed. Results Of the 57 patients, conversion to open surgery was required in 2 patients, and major venous resection and reconstruction were performed in 9 patients, including 8 end-to-end anastomosis, and 1 synthetic graft interposition. Total pancreatectomy was carried out in 4 patients. For the remaining 53 patients, pancreaticojejunal mucosal anastomoses were carried out in 50 patients, and sleeve pancreaticojeju-nostomy in 3 patients. The mean operative time and operative blood loss were 497 (240～720) min and 435 (50～3 000 ) ml, respectively. The mean postoperative hospital stay was 17. 7 ( 6. 0 ～59. 0 ) days. Postoperative complications were detected in 26. 3% (15/57) of patients, which included delayed gastric emptying (DGE) in 4 patients, Grade B pancreatic fistula (PF) in 4 patients, biliary fistula ( BF) in 2 patients, postpancreatectomy hemorrhage ( PPH) in 2 patients, intraabdominal infection in 1 patient and pulmonary infection in 2 patients. All the patients with DGE recovered with conservative treatment and they were discharged home. Reoperation was only required in the two patients with PPH. One patient died after the operation. The postoperative pathological results revealed pancreatic duct adenocarcinoma in 53 patients, adenosscale carcinoma in 1 patient and neuroendocrine carcinoma in 3 patients. The maximum and minimum tumor sizes were 7. 0 cm×5. 0 cm×3. 5 cm and 2. 5 cm×1. 5 ×1. 0 cm, respectively. The mean lymph nodes harvest and positive lymph node retrieval were 14(1～60) and 0. 7(0～3), respectively. Negative resection margins were obtained in 84. 2% (48/57) of patients. This study was censored on December 31, 2017. The follow-up for these patients ranged between 1 to 32 months. Mortality occurred in 21 patients, including 1 patient with a ruptured aneurysm 2 months after operation, 2 patients with GI bleeding 2 and 9 months respectively after operation, 1 patient with severe pulmonary infection and 17 patients with cancer recurrence with survival varying from 2 to 21 months. 35 patients were still alive. Conclusion Laparoscopic pancreaticoduodenectomy is a safe and feasible procedure for pancreatic head cancer.

Objective To explore the value of endoscopic retrograde cholangiopancreatography (ERCP) in patients with choledocholithiasis after Billroth Ⅱ gastrointestinal anastomosis.Methods A retrospective cohort study was conducted to retrospective review the data of 189 patients with choledocholithiasis treated by ERCP from December 2015 to November 2017 in Department of Hepatobiliary Surgery,Second Hospital of Hebei Medical University.According to the history of Billroth Ⅱ gastrointestinal anastomosis,the patients who have not undergone digestive surgery were divided into the normal group(n =167) and patients who have undergone digestive surgery were divided into reconstruction group (n =22).The operation time,the success rate of stone extraction,complications,the hospitalization time,total hospitalization expenses were compared between the two groups.Measurement data with normal distribution were represented as (Mean ± SD) and comparison between groups was analyzed using the t test.Measurement data with skewed distribution were described as M (range).Comparisons of count data were analyzed using the x2 test or Fisher exact probability.Results The operation time of the normal group was (40.18 ± 11.80) min,and the success rate of ERCP was 97.60％ (163/167),the operation time of reconstruction group was(61.81 ± 13.21) min,and the success rate of ERCP was 81.82％ (18/22),There were significant differences between the two groups (t =0.105,x2 =10.400,P ＜ 0.05).The complications,the hospitalization time and the total hospitalization expense of the normal group were 16.17％ (27/167),(3.47 ± 1.55) d,(20 620.69 ± 3 117.88) yuan,the reconstruction group were 18.18％ (4/22),(4.18 ± 2.08) d,(22 426.41 ±5 916.30) yuan,with no statistically significant difference (x2 =0.000,t =4.204),t =10.828,P ＞ 0.05).Conclusions ERCP is safe and feasible for patients with choledocholithiasis after Billroth Ⅱ gastrointestinal anastomosis.It also has a high success rate of stone removal and small trauma,which is worthy of promotion.

Objective To analyze the clinical outcomes and surgical procedures of 32 patients who underwent laparoscopic radical resection of hilar cholangiocarcinoma.Methods From January 2013 to July 2018,32 patients who were diagnosed to have hilar cholangiocarcinoma underwent total laparoscopic treatment in Second Hospital of Hebei Medical University.The clinical data of these patients were recorded,including the general data,Bismuth types,AJCC types,postoperative complications,pathological findings,and follow-up results.Results This study included 20 males and 12 females with a mean age of 60.9±8.8 years and a body mass index of 22.6±3.2 kg/m2.According to the preoperative imaging studies,the Bismuth types Ⅰ,Ⅱ,Ⅲa,Ⅲb,and Ⅳ were found in 12,2,3,4 and 11 patients,respectively.Laparoscopic radical resection of hilar cholangiocarcinoma and bilioenteric anastomosis was performed in 12 patients,with radical resection and external bile drainage in 6 patients,extended hemihepatectomy with caudate lobectomy in 6 patients and concomitant portal vein resection in 2 patients.The mean operative time was 365.6± 121.9min and the median intraoperative blood loss was 300 (75,400) ml.Intraoperative red cell and plasma transfusion were 0-15 U and 400(0,625)ml,respectively.According to the Clavien-Dindo complication classification system,5 of 32 (15.6％) patients developed type Ⅱ morbidity.The postoperative pathological findings revealed bile duct adenocarcinoma in 30 patients and mucinous adenocarcinoma in 2 patients.The median size of cancer was 3.0 (1.0,3.5) cm.According to the 8th AJCC staging system,stage Ⅰ,Ⅱ,Ⅲ,and Ⅳ were found in 6,13,11,and 2 patients,respectively.A negative resection margin was achieved in 24 of 32 patients (75％).Up to August 6,30 of 32 patients (93.8％) were followed up and the overall 1-,2-,and 3-year survival rates for the patients who underwent laparoscopic radical resection for hilar cholangiocarcinoma were 80.0％,53.0％,and 53.3％.The median survival time was 21.8 months.Conclusion Total laparoscopic surgery for hilar cholangiocarcinoma was safe and feasible if performed by an experienced surgeon after accurate preoperative evaluation.

Objectives: To propose a novel clinical classification system of gallbladder cancer, and to investigate the differences of clinicopathological characteristics and prognosis based on patients who underwent radical resection with different types of gallbladder cancer. Methods: The clinical data of 1 059 patients with gallbladder cancer underwent radical resection in 12 institutions in China from January 2013 to December 2017 were retrospectively collected and analyzed.There were 389 males and 670 females, aged (62.0±10.5)years(range:22-88 years).According to the location of tumor and the mode of invasion,the tumors were divided into peritoneal type, hepatic type, hepatic hilum type and mixed type, the surgical procedures were divided into regional radical resection and extended radical resection.The correlation between different types and T stage, N stage, vascular invasion, neural invasion, median survival time and surgical procedures were analyzed.Rates were compared by χ² test, survival analysis was carried by Kaplan-Meier and Log-rank test. Results: Regional radical resection was performed in 940 cases,including 81 cases in T1 stage,859 cases in T2-T4 stage,119 cases underwent extended radical resection;R0 resection was achieved in 990 cases(93.5%).The overall median survival time was 28 months.There were 81 patients in Tis-T1 stage and 978 patients in T2-T4 stage.The classification of gallbladder cancer in patients with T2-T4 stage: 345 cases(35.3%)of peritoneal type, 331 cases(33.8%) of hepatic type, 122 cases(12.5%) of hepatic hilum type and 180 cases(18.4%) of mixed type.T stage(χ²=288.60,P<0.01),N stage(χ²=68.10, P<0.01), vascular invasion(χ²=128.70, P<0.01)and neural invasion(χ²=54.30, P<0.01)were significantly correlated with the classification.The median survival time of peritoneal type,hepatic type,hepatic hilum type and mixed type was 48 months,21 months,16 months and 11 months,respectively(χ²=80.60,P<0.01).There was no significant difference in median survival time between regional radical resection and extended radical resection in the peritoneal type,hepatic type,hepatic hilum type and mixed type(all P>0.05). Conclusion With application of new clinical classification, different types of gallbladder cancer are proved to be correlated with TNM stage, malignant biological behavior and prognosis, which will facilitate us in preoperative evaluation,surgical planning and prognosis evaluation.