Objective:To explore the clinical data of patients with metastatic pheochromocytoma/paraganglioma (PPGL).Methods:The clinical data of 57 patients with metastatic pheochromocytoma/paraganglioma diagnosed and treated at Peking Union Medical College Hospital from January 2016 to June 2022 were reviewed, including 28 cases of pheochromocytoma(PCC) and 29 cases of paraganglioma(PGL). The clinical manifestations, biochemical indexes, tumour characteristics, and metastatic characteristics of the 57 patients were analysed.Results:There were 34 males and 23 females. The median age at the time of initial diagnosis was 34 (20, 54) years, 17 (29.3%) presented with concurrent metastases, and 40 (70.7%) with heterochronous metastases. The median time to presentation of metastases was 2.2 (0, 5.0) years (range 0-22 years). Adrenergic symptoms were present in 45 cases (78.6%) at the time of initial diagnosis, and the median size of the primary tumour was 6.7 (5.0, 9.0) cm. Excessive catecholamine secretion was present in 48 cases (81.4%). The most common locations of metastasis were lymph nodes (71.9%, 41/57), bone (47.3%, 27/57), lung (38.6%, 20/57), and liver (35.1%, 20/57). Metastatic PGL had more multifocal metastases than PCC [10 (34.5%) vs. 2 (7.1%), P=0.011)], was more frequently associated with SDHB mutations [13 (42.9%) vs. 3 (10.7%), P=0.008], and was more likely to have concurrent metastases [12 (41.3%) vs. 5 (17.9%), P=0.005]. Metastatic PCC primary tumours were larger compared to PGL [median length 8.9 (4.0, 17.0) cm vs. 6.1 (1.0, 15.8) cm, P=0.020]. Conclusions:Patients with PGL present with metastases over an extremely wide time span, and patients diagnosed with PPGL should be followed throughout their lives. PGL is more prone to multifocal metastases and simultaneous metastases than PCC, and PGL is more highly correlated with the SDHB mutation.

Production internship is an important teaching tache for undergraduate students to carry out engineering training by using professional skills, and it is a key starting point for fostering application-oriented talents in biotechnology. The Course Group of 'production internship of biotechnology majors' of Binzhou University is investigating application-oriented transformation for local regular colleges and universities, as well as fostering high-level application-oriented talents. By taking green fluorescent protein (GFP) polyclonal antibody as an example, the reform and practice on teaching content, teaching mode, assessment method, continuous improvement of curriculum were carried out. Moreover, the characteristics of the Yellow River Delta-Binzhou Biotechnology & Pharmaceutical Industrial Cluster were taken into account to intensify academic-enterprise cooperation. On one hand, this Course Group designed and rearranged the course contents, carried out essential training through online resources and platforms such as virtual simulation, and recorded, tracked and monitored the progress of production internship through practical testing and software platforms like 'Alumni State'. On the other hand, this Course Group established a practice-and application-oriented assessment method in the process of production internship and a dual evaluation model for continuous improvement. These reform and practices have promoted the training of application-oriented talents in biotechnology, and may serve as a reference for similar courses.
Humans ; Internship and Residency ; Curriculum ; Students ; Biotechnology

Cytokine (CK) is a low molecular weight soluble protein produced by a variety of cells induced by immunogen, mitogen or other stimulants. CK is capable of transmitting information between cells and having functions of immune regulation and effect. At present, common detection methods for cytokines include bioactivity detection, immunological detection and molecular biology method. Cytokines are involved in the occurrence and development of numerous autoimmune diseases, which can not only be used as an important biomarker of diseases, but also for the differential diagnosis of different autoimmune diseases, which is an important target for the clinical process monitoring and prognosis of diseases. In addition, antibodies and small molecule inhibitors targeting cytokines are gradually applied to the clinic, which also provides a safer and more effective treatment strategy for autoimmune diseases. This article reviews the application value of cytokines in autoimmune diseases.

Objective:To study the effect of left atrioventricular interphase (LAVI) via esophageal electrocardiogram on cardiac function after dual-chamber pacemaker implantation in patients with high-degree atrioventricular block.Methods:Using a prospective approach, 40 patients with high-degree atrioventricular block who would undergo dual-chamber pacemaker implantation from January 2017 to March 2018 in Department of Cardiovascular Medicine, Dalian Municipal Central Hospital Affiliated of Dalian Medical University were enrolled. All patients accepted esophageal electrocardiogram tests at 3 months after the implantation, to exam the interatrial conduction time (IACT) of sinus rhythm and pacing rhythm, and interventricular conduction time (IVCT). Then based on the outcome of the echocardiography test, the optimal atrioventricular delay (AVD) of the pacemaker of each patient was determined while the LAVI differed from 100 ms to 150 ms. The left ventricular ejection fraction (LVEF), peak speed of blood flow velocity in early mitral orifice diastole (E), E peak deceleration time (EDT), peak speed of early mitral annular diastolic movement (e′), isovolumic relaxation time (IVRT) and left atrial volume (LAV) were tested by echocardiogram before implantation, before AVD adjustment at 3 months after implantation, after AVD adjustment at 3 months after implantation, and 6, 12, and 18 months after implantation. Then, the left atrial volume index (LAV/body surface area) and E/e′ were calculated.Results:Among the 40 patients, the IACT of sinus rhythm was (55.55 ± 10.33) ms, the IACT of pacing rhythm was (93.95 ± 12.77) ms, and the mean IVCT was (63.20 ± 17.84) ms; the optimal LAVI was 110 to 150 (132.00 ± 10.43) ms, and notably, the optimal LAVI between 120 and 140 ms was 82.5% (33/40). The LVEF, EDT, IVRT, left atrial volume index and E/e′ from before AVD adjustment of 3 months after implantation to follow-up endpoint (18 months after implantation) were significantly improved compared with those before implantation, and there were statistical differences ( P<0.01); the EDT and IVRT after AVD adjustment at 3 months after implantation were significantly improved than those before AVD adjustment at 3 months after implantation: (142.15 ± 35.58) ms vs. (125.94 ± 31.13) ms and (119.52 ± 22.15) ms vs. (133.92 ± 23.87) ms, and there were statistical differences ( P<0.05); the IVRT and left atrial volume index 18 months after implantation were significantly improved compared with those before AVD adjustment at 3 months after implantation: (122.07 ± 16.99) ms vs. (133.92 ± 23.87) and 32.94 ± 3.22 vs. 35.43 ± 5.76, and there were statistical differences ( P<0.05). Conclusions:Optimizing the LAVI after dual-chamber pacemaker implantation via esophageal electrocardiogram can improve the long-term prognosis of patients with high-degree atrioventricular block.

OBJECTIVE：To in vestigate the effects of dexmedetomidine （Dex）on SIRT 1/Akt/GSK3β/β-catenin signaling pathway in cerebral injury of sepsis model rats ，and explore the mechanism of its protecitve effect on cerebral injury. METHODS ： A total of 80 male SD rats were randomly divided into sham operation group （Sham group ），sepsis group （CLP group ），CLP+Dex group（10 μg/kg Dex），CLP+Dex+Sirtinol group （10 μg/kg Dex+2 μL/100 g SIRT 1 inhibitor sirtinol ），with 20 mice in each group. Two hours before modeling ，CLP+Dex+Sirtinol group was injected with sirtinol via lateral ventricle. Sepsis model was induced by cecal ligation and perforation in each group （in sham group ，only operation was performed but no ligation was performed）. At 0，3，6 h after modeling ，CLP+Dex group and CLP+Dex+Sirtinol group were given Dex （10 μg/kg） intraperitoneally，Sham group and CLP group were given constant volume of normal saline intraperitoneally. Cerebral tissue water content，Evans blue （EB）content，apoptosis in cerebral cortex ，the levels of IL- 1β and TNF-α in cerebral tissue as well as the protein expression of SIRT 1，p-Akt，p-GSK3β and β-catenin in hippocampus were detected 24 h after last medication. RESULTS ： Compared with Sham group ，cerebral tissue water content ，EB content ，the number of apoptotic cells in cerebral cortex as well as the levels of IL- 1β and TNF-α in cerebral tissue were increased significantly（P＜0.05），while the protein expression of SIRT 1， p-Akt，p-GSK3β and β-catenin in hippocampus were decreased significantly （P＜0.05）. Compared with CLP group ，cerebral tissue water content ，EB content ，the number of apoptotic cells in cerebral cortex as well as the levels of IL- 1β and TNF-α in cerebral tissue were decreased significantly in CLP+Dex group （P＜0.05），while the protein expression of SIRT 1，p-Akt，p-GSK3β and β-catenin in hippocampus were increased significantly （P＜0.05）. Sirtinol could significantly reverse the above-mentioned cerebral protection and factor regulation effects of Dex （P＜0.05）. CONCLUSIONS ：Dex can protect the cerebral tissue of sepsis model rats，which may play an anti-inflammatory and anti-apoptotic role by activating SIRT 1/Akt/GSK3β/β-catenin signaling pathway ，so as to reduce cerebral edema ，protect blood-brain barrier and reduce cerebral injury.

Objective: To investigate the clinical features and pathogenic genes of a family with osteosclerosis. Methods: Six patients and six family members from a family in Jiangsu were tested for biochemical parameters, bone metabolic markers, bone mineral density, thoracolumbar anterior lateral slices, skull positive lateral radiographs, and pelvic plain films. Meanwhile, Sanger sequencing was performed to detect gene mutations of the proband and five other family members with high bone mass. The conformation of the mutational low-density lipoprotein receptor-related protein 5 (LRP5) protein was predicted by SWISS-MODEL. Results: Four adult patients (one male and three females) were tall, with mandibular enlargement and kyphosis in the center of the lower jaw, and none of the four had fractures. Their X ray examination revealed that the skull and long bone cortex was thickened, while the sella and mandible was enlarged. In addition, the absolute values of bone mineral density at each site of all patients were significantly higher as compared with the standard age- and sex-matched adults or adolescent mean reference values, with Z scores of L2-4, femoral neck and total hip being (6.31±4.03) SD, (6.56±2.36) SD, and (7.19±2.03) SD, respectively. The results of genetic sequencing revealed that all six patients carried a heterozygous mutation (c.331G>T; D111Y) in exon 2 of LRP5 gene, while other family members showed wild type (c.331G>G; D111D). Functional prediction indicated that this mutation was located at the amino acid terminal of exon 2 of LRP5 gene, which encodes the first β-helix-generating region of LRP5 protein. Conclusion The D111Y mutation in LRP5 gene leads to a clinical phenotype characterized by benign increased bone mineral density without increasing the risk of fracture. This mutation may further affect the downstream Wnt signaling pathway by altering the spatial structure of LRP5 protein, thereby promoting maturation and differentiation of osteoblasts and resulting in osteosclerosis.

Objective To observe the short-term clinical outcomes of arthroscopic single-tibial tunnel Pushlock fixation in the treatment of tibial avulsion fracture of the posterior cruciate ligament (PCL).Methods From December 2015 to January 2018,15 patients with tibial avulsion fracture of PCL were treated at Department of Articular Surgery,The Third Affiliated Hospital to Hebei Medical University.They were treated with arthroscopic single-tibial tunnel Pushlock fixation.All the fractures were fresh.They were 9 males and 6 females,aged from 14 to 64 years (average,33.6 years).The short-term outcomes were evaluated by comparing their preoperative and postoperative flexion of the knee,International Knee Documentation Committee (IKDC) and Lysholm scores.Results The operation lasted from 40 to 70 minutes (average,53 minutes).The 15 patients were successfully followed up for 10 to 18 months (average,12.2 months).All the fractures united well within 3 months after operation.All the posterior drawer tests were negative,showing no displacements or no complications like lesions of popliteal fossa,nerves and vessels,or impaired knee extension.At 6 months after surgery,the knee flexion (123.4°± 6.5°),the Lysholm scores (91.8 ± 3.5) and the IKDC scores (95.5 ± 1.6) were all significantly improved compared with the preoperative values (77.5°±13.1°,46.8 ±8.9 and 37.0±8.9) (P ＜0.05).Conclusion The tibial avulsion fracture of PCL can be treated by arthroscopic single-tibial tunnel Pushlock fixation with satisfactory early outcomes.

Objective To compare the short-term outcomes between lateral retinacular release (LRR) via a prolonged incision versus arthroscopic LRR in osteofascial medial patellofemoral ligament reconstruction (MPFL-R) for recurrent patellofemoral dislocation.Methods From May 2012 to March 2015,51 patients with recurrent patellofemoral dislocation were treated at the Department of Joint Surgery,The Third Affiliated Hospital to Hebei Medical University.They were 16 males and 35 females,aged from 20 to 36 years (mean 27.2 years).Of them,28 underwent LRR via a prolonged incision in MPFL-R (incision group) and 23 did arthroscopic LRR in MPFL-R (microscopic group).Imaging changes were assessed by comparing preoperative and postoperative values of patella extensibility,patellar tilt angle,trochlear groove angle and tibial tuberosity-trochlear groove spacing (TT-TG);the knee joints were scored with Kujala and Lysholm functional rating systems at the last follow-ups.Results The 2 groups were compatible due to insignificant differences in the general data between them preoperatively (P ＞ 0.05).The 51 patients were followed up for 10 to 32 months (average,20 months).The rate of dislocation in the incision and microscopic groups were 0 and 8.7％ (2/23),respectively.There were no such complications in either group as patellar fracture,deep venous thrombosis of the lower extremity or limited joint extension or flexion.The last follow-ups revealed that no patients in either group were inflicted with patellofemoral re-dislocation.In the incision group,the patella extensibility,patellar tilt angle,Kujala score and Lysholm score were,respectively,19.65％ ± 0.75％,12.39°±0.76°,56.37 ±2.94 points and 51.64 ±6.22 points preoperatively,and 10.34％ ± 0.60％,8.83°±0.89°,92.68 ±2.75,and 90.71 ± 1.91 points postoperatively;in the microscopic group,the patella extensibility,patellar tilt angle,Kujala score and Lysholm score were,respectively,19.56％ ±0.62％,12.35° ± 0.66°,57.46 ± 3.08 points and 52.20 ± 6.94 points preoperatively,and 10.24％ ±0.66％,8.78° ± 0.92°,92.53 ± 3.32 points and 90.41 ± 2.90 points postoperatively.There were significant differences between the preoperative and postoperative values in both 2 groups (P ＜ 0.05).However,there were no significant differences between the preoperative and the postoperative values in trochlear groove angle or TT-TG in either group (P ＞ 0.05).There were no significant differences between the 2 groups in all the above preoperative and postoperative values (P ＞ 0.05).Conclusion In MPFL-R for recurrent patellofemoral dislocation,both LRR via a prolonged incision and arthroscopic LRR can improve the patellofemoral joint alignment and stability of the tibia,leading to good short-term outcomes.

Bacterial ghosts are bacterial cell envelopes devoid of cytoplasmic contents while maintaining their cellular morphology, which can be used as a new vaccine and delivery vector. In this study, a clinical isolate of avian pathogenic Escherichia coli (APEC) strain DE17 was used to prepare bacterial ghost through three different ways. The results showed that the cleavage efficiency of DE17 bacterial ghost was 99.9% with the lysis plasmid containing the PhiX174 lysis gene E. Scanning electron microscopy showed that transmembrane tunnels were formed in the middle or both ends of the cell envelope of DE17. Furthermore, the DE17 bacterial ghost was prepared with one of cell penetrating peptides (CPPs) named MAP (KLALKLALKALKAALKLA), which will completely inactivate DE17 (OD₆₀₀=0.1) by 10 μmol/L MAP. The cell envelope showed a gully-like structure and obvious transmembrane tunnels were not found through the SEM. However, the DE17 could not be lysed by importing the lysis plasmid (pBV220-MAP), which was used to express MAP. The present study will benefit for research on bacterial ghost preparation methods and provide a reference for biosafety of bacterial ghost vaccines.

Objective To isolate and identify anti-tumor constituents from Lysimachia christinae Hance.Methods The compounds were isolated and purified by chromategraphy on AB-8 macroporous resin, silica gel, Sephadex LH-20, ODS and preparative HPLC.Their structures were elucidated using chemical and spectroscopic methods.All thecompounds obtained were evaluated on their growth inhibitory effects in vitro using the MTT method.Results Eleven compounds were isolated from the whole plant of L.christinae Hance and were identified as 3,3'-dimethoxy-6,6'-di[(Z)-pentadec-10-en-1-yl]-[1,1'-bi(cyclohexane)]-3,3',6,6'-tetraene-2,2',5,5'-tetraone(1),physcion(2),munjistin(3), 9,16-dioxooctadec-10,12,14-trienoic acid(4),cyclamiretinA-3-O-{β-D-xylopyranosyl-(1→2)-β-D-glucopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→4)]-α-L-arabinopyranoside(5),lysikoianoside(6),quercetin-3-O-glucopyranoside(7),isorhamnetin3-O-β-D-galactopyranoside(8),3',4',5',5,7-pentahydroxyflavone(9), rutin(10), and quercetin(11).The screening results of antitumor activity in vitro showed that IC50 of compound 5 for human cervical carcinoma cells HeLa, human osteosarcoma cells U20S, human lung adenocarcinoma cells PC-9,and human colorectal carcinoma cells CT-26 was 2.29, 1.22, 1.43, and 1.29 μmol/L respectively.Conclusion Compound 1 is a new compound and compounds 2-4 are isolated from the plants of Lysimachia for the first time.Compound 5 shows obvious inhibitory effects on tumor cell growth in vitro.