ObjectiveTo construct and evaluate a mouse model that simulates the pathological progression of immune checkpoint inhibitor-associated pneumonitis (CIP), in order to address the critical issue of the lack of standardized animal models in this research field. MethodsThirty-five SPF-grade 6-week-old male C57BL/6J mice were randomly assigned to a control group (n=5), a bleomycin (BLM) group (n=15), and a BLM+programmed death-1 (PD-1) monoclonal antibody (BLM+PD-1) group (n=15). Corresponding drugs were administered via intraperitoneal injection for intervention. The general condition, body weight changes, survival rate, and pulmonary histopathological alterations were systematically monitored. ResultsCompared with the BLM group, the BLM+PD-1 group exhibited significantly more severe pulmonary fibrosis, characterized by progressive body weight loss and more severe deterioration of general conditions. Histopathological examination showed that pulmonary inflammation was alleviated 4 weeks post-modeling in the BLM group. In contrast, pulmonary lesions continued to worsen in the BLM+PD-1 group, with no evidence of resolution of inflammation 28 days post-modeling. Quantitative immunohistochemical analysis revealed that the positive expression rate of interleukin-17A (IL-17A) in lung tissues reached 34.28% in the BLM+PD-1 group 14 days post-modeling, which was markedly higher than that in the BLM group (24.02%, P < 0.000 1). The positive expression rate of α-smooth muscle actin (α-SMA) was 12.40% in the BLM+PD-1 group 14 days post-modeling, nearly twice the level in the BLM group (5.87%). 28 days post-modeling, the positive expression rate of α-SMA remained at 7.72% in the BLM+PD-1 group, while it declined to 3.38% in the BLM group. ConclusionThe addition of a PD-1 inhibitor to bleomycin treatment accelerates the progression of pulmonary interstitial fibrosis and effectively recapitulates the pathological process of lung injury and fibrosis driven by aberrant immune activation during clinical immunotherapy. Accordingly, this study successfully establishes a dynamic CIP animal model recapitulating the progression from early inflammatory response to pulmonary fibrosis. This model addresses the critical shortage of animal models for this condition and offers an experimental platform for further mechanistic studies and the development of therapeutic strategies.

Brain tinnitus is a common clinical condition whose pathogenesis is not yet fully understood,and it still lacks effective therapies in clinical practice.Professor Liu Maocai believes that the clinical manifestations of brain tinnitus are usually characterized by deficiency syndrome,and sometimes are complicated excess syndrome.The disease is located in the brain and is closely related to the liver,spleen,and kidney.The treatment of brain tinnitus should be focused on tonifying the liver and kidney,and strengthening the spleen,and methods of calming the liver to extinguish wind,resolving phlegm to open the orifices,and activating blood circulation to unblock collaterals can be used for excess syndrome.In clinical practice,Professor Liu Maocai is skilled in using herb pairs such as Astragali Radix-Codonopsis Radix,Eucommiae Cortex-Achyranthis Bidentatae Radix,Paeoniae Radix Alba-Bupleuri Radix,Angelicae Sinensis Radix-Chuanxiong Rhizoma,Gastrodiae Rhizoma-Uncariae Ramulus cum Uncis,and Acori Tatarinowii Rhizoma-Polygalae Radix.Professor Liu Maocai's experience in treating brain tinnitus from the perspective of the liver,spleen,and kidney can provide valuable references for the clinical diagnosis and treatment of brain tinnitus with traditional Chinese medicine.

Objective:To investigate the acute gastrointestinal injury (AGI) in patients with extracorporeal membrane oxygenation (ECMO) at the early stage of operation and its influencing factors.Methods:A total of 70 patients with ECMO who were hospitalized in the Emergency Care Unit of Guangxi Zhuang Autonomous Region People's Hospital from September 2020 to December 2021 were retrospectively analyzed, and a total of 70 patients with ECMO who were hospitalized in the emergency care unit of Guangxi Zhuang Autonomous Region People's Hospital from September 2020 to December 2021 were retrospectively analyzed. According to the 2012 guidelines of the European Society of Intensive Care Medicine on the classification of acute gastrointestinal injury in critically ill patients, the patients were divided into AGI group and non-AGI group. The incidence of acute gastrointestinal injury in the early stage was statistically analyzed, and the results of blood gas analysis during ECMO loading and ECMO parameters, hemodynamic indexes and biochemical indexes after ECMO transfer were statistically analyzed. To explore the influencing factors and independent risk factors of AGI in the early stage. In addition, 70 patients were divided into successful group and non-successful group according to whether they were successfully withdrawn. The occurrence of acute gastrointestinal injury between the two groups was compared, and the effect of acute gastrointestinal injury on ECMO patients was analyzed.Results:Among the 70 ECMO patients, the incidence of early AGI was 71.43% (50 cases), and the components of AGI Ⅰ, Ⅱ, Ⅲ and Ⅳ were 18.57% (13 cases), 41.43% (29 cases), 11.43% (8 cases) and 0% (0 cases), respectively. ① Univariate analysis showed that systolic blood pressure, diastolic blood pressure, mean arterial pressure (MAP), vasoactive drug index (VIS), pH, lactic acid and BMI were significantly different between AGI group and non-AGI group when ECMO was used ( P < 0.05). Logistic binary regression analysis showed that BMI was an independent risk factor for early AGI in ECMO patients (ROC area 0.657, 95% confidence interval 0.522-0.791 ( P < 0.05), and Yoden index 0.15). (3) The AGI composition ratio of the unsuccessful group was higher than that of the unsuccessful group ( P < 0.05). Conclusions:Patients with ECMO have a high incidence of AGI in the early stage, mainly occurring in grade I and Ⅱ. Systolic blood pressure, diastolic blood pressure, MAP, VIS, pH, lactic acid and BMI when ECMO is put on are influential factors for the early development of AGI in ECMO patients, among which BMI is an independent risk factor for the early development of AGI in ECMO patients. The occurrence of AGI reduces the probability of successful withdrawal in ECMO patients.

Objective:To investigate the clinical, skeletal muscle pathological, and genetic characteristics of fatal infantile hypertonic myofibrillar myopathy (FIHMM).Methods:The clinical manifestations, laboratory assessments data and gene sequencing results of 10 patients diagnosed with FIHMM in Shenzhen Children′s Hospital from February 2017 to April 2021 were retrospectively analyzed.Magnetic resonance imaging (MRI) of both musculoskeletal system and the brain, and electromyogram (EMG) were performed in 3 cases, while muscle biopsy was performed in 2 cases.Results:Among these 10 cases, 1 case was from Northeast China and 1 case from East China, while the rest 8 cases were from South China.Eight of the 10 patients were male, and the other 2 cases were female.They were all born normal and not related to each other.The age of onset varied from 2 to 12 months.The main clinical manifestations for all the patients were progressive rigidity of the rectus abdominis (8 cases), neck muscles (7 cases), rectus abdominis (2 cases) and intercostal muscles (1 case), resulting in respiratory failure.Mildly to moderately elevated serum creatine kinase level was detected (436-5 804 IU/L) (reference range: 24-229 IU/L). Complex repetitive discharges can be seen in the EMG, without any myotonic potential.Muscle fiber degeneration, necrosis, and vacuolar degeneration were noted in the histopathological examination of the vastus lateralis and rectus abdominis.An abnormal red granular deposit was observed in a portion of the field of the modified Gomory Trichrome staining.Immunohistochemistry showed substantial deposition of desmin.Under the electron microscopy, the sarcomere structure of the muscle fibers was seriously disordered, with the destruction of Z-bands and the presence of granular deposits.The whole-exome sequencing identified the same homozygous variation c. 3G>A, p.Met1? of CRYAB gene in all the patients, but heterozygous variation in their parents. Conclusions:Axial muscles involvement, such as rectus abdominis rigidity, is the main clinical characteristic of FIHMM.c.3G>A, p.Met1? mutation in the CRYAB gene is a hotspot mutation in Chinese children.

Objective:To establish autoverification rules for coagulation tests in multicenter cooperative units, in order to reduce workload for manual review of suspected results and shorten turnaround time (TAT) of test reports, while ensure the accuracy of results.Methods:A total of 14 394 blood samples were collected from fourteen hospitals during December 2019 to March 2020. These samples included: Rules Establishment Group 11 230 cases, including 1 182 cases for Delta check rules; Rules Validation Group 3 164 cases, including 487cases for Delta check; Clinical Application Trial Group 77 269 cases. Samples were analyzed for coagulation tests using Sysmex CS series automatic coagulation analyzers, and the clinical information, instrument parameters, test results, clinical diagnosis, medication history of anticoagulant and other relative results such as HCT, TG, TBIL, DBIL were summarized; on the basis of historical data, the 2.5 and 97.5 percentile of all data arranged from low to high were initially accumulated; on the basis of clinical suggestions, critical values and specific drug use as well as relative guidelines, autoverification rules and limits were established.The rules were then input into middleware, in which Stage I/Stage II validation was done. Positive coincidence, negative coincidence, false negative, false positive, autoverification pass rate, passing accuracy (coincidence of autoverification and manual verification) were calculated. Autoverification rules underwent trial application in coagulation results reports.Results:(1) The autoverification algorisms involve 33 rules regarding PT/INR, APTT, FBG, D-dimer, FDP,Delta check, reaction curve and sample abnormalities; (2)Autoverification Establishment Group showed autoverification pass rate was 68.42% (7 684/11 230), the false negative rate was 0%(0/11230), coincidence of autoverification and manual verification was 98.51%(11 063/11 230), in which positive coincidence and negative coincidence were respectively 30.09% (3 379/11 230) and 68.42%(7 684/11 230); Autoverification Validation Group showed autoverification pass rate was 60.37%(1 910/3 164), the false negative rate was 0%(0/11 230), coincidence of autoverification and manual verification was 97.79%(3 094/3 164), in which positive coincidence and negative coincidence were respectively 37.42%(1 184/3 164) and 60.37%(1 910/3 164); (3) Trialed implementation of these autoverification rules on 77 269 coagulation samples showed that the average TAT shortened by 8.5 min-83.1 min.Conclusions:This study established 33 autoverification rules in coagulation tests. Validation showedthese rules could ensure test quality while shortening TAT and lighten manual workload.

Objective: To establish a set of rules for autoverification of blood analysis, in order to provide a way to validate autoverification rules for different analytical systems, which can ensure the accuracy of test results as well as shorten turnaround time (TAT) of test reports. Methods: A total of 34 629 EDTA-K2 anticoagulated blood samples were collected from multicenter cooperative units including the First Hospital of Jinlin University during January 2017 to November 2017. These samples included: 3 478 cases in Autoverification Establishment Group, including 288 cases for Delta check rules; 5 362 cases in Autoverification Validation Group, including 2 494 cases for Delta check; 25 789 cases in Clinical Application Trial Group. All these samples were analyzed for blood routine tests using Sysmex XN series automatic blood analyzers.Blood smears, staining and microscopic examination were done for each sample; then the clinical information, instrument parameters, test results and microscopic results were summarized; screening and determination of autoverification conditions including parameters and cutoff values were done using statistical analysis. The autoverification rules were input into Sysmex Laboman software and undergone stage Ⅰ validation using simulated data, and stage Ⅱ validation for post-analytical samples successively. True negative, false negative, true positive, false positive, autoverification pass rate and passing accuracy were calculated. Autoverification rules were applied to autoverification blood routine results and missed detection rates were validated, and also data of autoverification pass rate and TAT were obtained. Results: (1)The selected autoverification conditions and cutoff values included 43 rules involving WBC, RBC, PLT, Delta check and abnormal characteristics. (2)Validation of 3 190 cases in Autoverification Establishment Group showed the false negative rate was 1.94%(62/3 190)(P<0.001), autoverification pass rate was 76.74%, passing accuracy was 97.47%; Validation of 2 868 cases in Autoverification Validation Group, the false negative rate was 3.38%(97/2 868)(P=0.002), autoverification pass rate was 42.26%, passing accuracy was 92.00%; Validation of Delta check on 288 cases in Autoverification Establishment Group and 2 494 cases in Autoverification Validation Group showed the false negative rates were respectively 1.39% and 2.61%(P<0.001). (3)Three hospitals adopted these rules of autoverification for 25 789 blood routine samples, and found that the average TAT of blood routine test reports were shortened by 24min, 32min and 7min respectively, the rate of samples reported within 30min were elevated by 33%, 53% and 7%. The autoverification pass rates were 72%-74%. Conclusions The application of this set of 43 autoverification rules in blood sample analysis can ensure test quality while shortenTAT and improve work efficiency. It is worth pointing out that for the same analytical systems in this research, validation is necessary before application of this set of rules, and periodic validation is required during application to make necessary adjustment; for different analytical systems, as this research provide a way to establish autoverification rules for blood routine tests.Clinical labs may establish their own suitable autoverification rules on the basis of technological parameters. (Chin J Lab Med, 2018, 41: 601-607)

Objective To investigate the preparation of a polycaprolactone (PCL)/poly (lactic acid-glycolic acid) (PLGA) degradable ureteral stent and its degradation ability in vitro.Methods From December 2010 to September 2012,degradable ureteral stent was fabricated by electrospinning,using different concentration of PCL/PLGA (5％,10％,15％,20％,25％,30％).The structure of stent was scanned by electron microscopy (SEM).The Instron system was used to test the mechanical property of those stents.The PCL/PLGA stents of different concentration (5％,15％,25％) were immerged into the urine to assess their degradable ability 7,14,21,28,35,42,49,56 days later.Results The inner diameter of the white tubular scaffold was 1.5 mm and the outer diameter was 2.0 mm.The SEM results showed that the scaffold had been made by nanofiber with the structure of multi-porous.The tension test showed that the mechanical property was enhanced with the increasing in the proportion of the PCL.The lowest stress at break was found in 5％ PCL/PLGA stent,while it was still sufficient for the mechanical criteria of degradable biomaterial.The degradation curves of different ratio of PCL/PLGA stent were close to a straight line.The 5％ PCL/PLGA stent collapsed into pieces within 28 d.The 15％ and 25％ PCL/PLGA collapsed within 42 d and 56 d,respectively.Conclusion The PCL/PLGA biodegradable ureteral stent has a good mechanical property and the degradation time can fully meet the demand of a ureteral stent.

Objective The present study was designed to investigate the risk factors affecting bleeding during percutaneous nephrolithotomy. Methods The records of 218 patients with percutaneous nephrolithotomy procedure by a single surgeon were retrospectively reviewed.The mean age was 48 years ( range,19 -70).One hundred and forty six patients had staghore stones,and 7 patients had previous open or percutaneous nephrolithotomy histories.Forty-one patients had concomitant diabetes mellitus,and 89 cases had hypertension.The following factors including age,sex,BMI,diabetes status,hypertension status,stone type,calix of puncture,previous open or percutaneous nephrolithotomy history,number of accesses,size of accesses,operative time,and surgeon experience were analyzed.Univariate analysis and multivariate step regression analysis were used for statistical assessment. Results 207 procedures were successfully performed,and 11 patients failed because of difficulty to establish the accesses.Single-tract was used in 176 cases and multiple-tract was used in 31 cases.163 cases were performed via a 18 F access and 44 cases via a 24 F access.The mean operative time was 78.4 min.The overall blood transfusion rate was 7.7％,and stone type ( P =0.034),diabetes ( P =0.030),number of accesses ( P =0.019 ),size of accesses ( P =0.008) and operative time (P =0.001 ) were the risk factors affecting blood transfusion requirement.The average hemoglobin (Hb) drop after percutaneous nephrolithotomy procedures was 11.2 g/L,and stone type ( P ＜ 0.001 ),diabetes ( P =0.015 ),number of accesses ( P =0.016),size of accesses ( P ＜ 0.001 ) and operative time ( P ＜ 0.001 ) were the risk factors affecting Hb drop.The following covariates including Hb drop:age,sex,BMI,previous open or percutaneous nephrolithotomy history,hypertension status,calix of puncture and surgeon experience were not risk factors affecting blood transfusion requirement and Hb drop.Multivariate stepwise regression analysis showed that diabetes ( OR =1.75 ),stone type ( OR =1.92),number of accesses ( OR =2.45 ),size of accesses ( OR =1.32) and operative time ( OR =1.66) significantly increased risk of bleeding. Conclusions Stone type,diabetes,number of accesses,size of accesses and operative time were the risk factors affecting blood loss during percutaneous nephrolithotomy.

Objective To study the expression of enhancer of zeste homolog 2 (EZH2) gene in transitional cell carcinoma (TCC) of the bladder celt lines, carcinoma tissues and normal bladder tis-sues and to evaluate the roles of EZH2 in the development and progression of bladder carcinoma. Methods RT-PCR, Western-blot and immunocytochemistry were used to analyze the expression of EZH2 of the bladder cell lines (T24, EJ, MGH-U1, BIU-87). The prostate cancer cell line PC-3M was used as an EZH2-positive cell line. EZH2 gene expressions in 45 cases of bladder carcinoma and 12 cases of normal bladder mueosa were detected by RT-PCR. Of cancer cases, 31 were superficial tumors and 14 were invasive tumors; 13 were G1, 21 were G2 and 11 were G3. Results EZH2 was detected in the 4 TCC cell lines. The EZH2 expression rate of TCC (82.2%) was significantly higher than that of normal bladder tissues (8.3%, P<0.05). The expression rate in superficial tumors was 74.2% and in invasive tumors was 100.0%, but there was no significant difference (P>0.05). The expression rates increased with tumor cell grade increase, but there was no significant difference (P> 0.05). Conclusions EZH2 could play an important role in the development and progression of blad-der carcinoma. It could be used as a potential gene therapy target of bladder cancer.

Objective To investigate the feasibility of gene modification of peripheral blood derived endothelial progenitor cells(EPCs).Methods The rabbit mononuclear cells were collected from rabbit peripheral blood and cultured.The uptaking test of DiI-Ac-LDL was directly visualized with fluorescent microscopy,the immunofluorescence analysis was performed to detect the expression of surface marker.And the human tissue kallikrein(HTK)gene was amplified with PCR,and inserted into pEGFP-N3 vector.EPCs were transfected with the constructed plasmid by means of lipidosome,and the HTK-EGFP fused protein was visualized directly with fluorescent microscopy,and the expression of HTK was detected by RT-PCR and ELISA.Results After 5～6 days of culture,spindle-shaped attached cells clustered together,and cobblestone-like cell layer appeared after 15 days.Cells were positive of uptaking DiI-Ac-LDL and expressing vWF and CD133 related antigen.Plasmids were correctly formed and expressed in the rabbit EPCs.The fused protein had activity of both HTK and EGFP.Conclusion The HTK gene modified EPCs is potential in the treatment of artery injury and endothelial dysfunction.