OBJECTIVE To observe the changes of serum proteins in the mice with carbapenem-resistant hyperviru-lent Klebsiella pneumoniae(CR-hvKP)bloodstream infection by using liquid chromatogram-mass spectrometer(LC-MS),find out the differentially expressed proteins and carry out corresponding biological function analysis.METHODS The models of ICR mice with CR-hvKP and classical Klebsiella pneumoniae(cKP)bloodstream infec-tions were established,the serum specimens were collected from the mice at 12 hours of establishment of the in-fection models and were detected by using LC-MS.The result of LC-MS was identified by using Maxquant soft-ware.The corresponding bioinformatics analysis was performed for the differentially expressed proteins.RESULTS As compared with the normal control group,there were 24 upregulated proteins and 20 downregulated proteins in CR-hvKP group.As compared with cKP group,there were 107 upregulated proteins in the CR-hvKP group.The 134 differentially expressed proteins were retrieved one by one from Uniprot database,it was found that the pro-teins were involved in biological regulation,immune process,biological interaction,movement,metabolic process,response to stimulation,signal transduction,inflammatory response,oxidative stress and angiogenesis.The signal pathway analysis involved multiple metabolism-related pathways,including complement and coagula-tion cascades,cholesterol metabolism,bacterial infection,heme clearance,and assembly,remodeling and clear-ance of plasma lipoprotein.In terms of the proteins being attached great importance,the expression levels of kal-likrein B1(KLKB1)and serpin family A(Serpina)1b were downregulated as compared with the normal control group and were upregulated as compared with the cKP group;the expression levels of serum amyloid A protein(SAA)1,SAA2,Vanin-3(VNN3)and hemoglobinβpolypeptide chain b2(HBB-b2)were upregulated as com-pared with both the cKP group and the normal control group.CONCLUSIONS The CR-hvKP bloodstream infection involves the activation and action of multiple proteins.The rise of SAA1,SAA2,VNN3 and HBB-b2 or the de-cline of KLKB1 and Serpina1b may indicate the CR-hvKP bloodstream infection.

OBJECTIVE To observe the changes of serum proteins in the mice with carbapenem-resistant hyperviru-lent Klebsiella pneumoniae(CR-hvKP)bloodstream infection by using liquid chromatogram-mass spectrometer(LC-MS),find out the differentially expressed proteins and carry out corresponding biological function analysis.METHODS The models of ICR mice with CR-hvKP and classical Klebsiella pneumoniae(cKP)bloodstream infec-tions were established,the serum specimens were collected from the mice at 12 hours of establishment of the in-fection models and were detected by using LC-MS.The result of LC-MS was identified by using Maxquant soft-ware.The corresponding bioinformatics analysis was performed for the differentially expressed proteins.RESULTS As compared with the normal control group,there were 24 upregulated proteins and 20 downregulated proteins in CR-hvKP group.As compared with cKP group,there were 107 upregulated proteins in the CR-hvKP group.The 134 differentially expressed proteins were retrieved one by one from Uniprot database,it was found that the pro-teins were involved in biological regulation,immune process,biological interaction,movement,metabolic process,response to stimulation,signal transduction,inflammatory response,oxidative stress and angiogenesis.The signal pathway analysis involved multiple metabolism-related pathways,including complement and coagula-tion cascades,cholesterol metabolism,bacterial infection,heme clearance,and assembly,remodeling and clear-ance of plasma lipoprotein.In terms of the proteins being attached great importance,the expression levels of kal-likrein B1(KLKB1)and serpin family A(Serpina)1b were downregulated as compared with the normal control group and were upregulated as compared with the cKP group;the expression levels of serum amyloid A protein(SAA)1,SAA2,Vanin-3(VNN3)and hemoglobinβpolypeptide chain b2(HBB-b2)were upregulated as com-pared with both the cKP group and the normal control group.CONCLUSIONS The CR-hvKP bloodstream infection involves the activation and action of multiple proteins.The rise of SAA1,SAA2,VNN3 and HBB-b2 or the de-cline of KLKB1 and Serpina1b may indicate the CR-hvKP bloodstream infection.

The traditional experimental teaching mode of forensic science and biological evidence is mostly confined to experimental operation, which is not capable of cultivating students' comprehensive quality and post competency. Therefore, it is urgent to seek an innovative teaching and training mode. At present, the experimental teaching of forensic science and biological evidence is dominated by teachers. There are some problems, such as insufficient training of students' scientific thinking and innovation ability, single teaching and evaluation model, and disconnection from the practical application. This paper proposes an experimental teaching design scheme of forensic science and biological evidence based on post competency training. The course is implanted in the framework of simulated cases, and the virtual simulation experiment platform and group discussion learning method are used to achieve a training model oriented by social needs and centered on students. In the preliminary study on the students who were trained in this mode of selected sections, we found that, compared with traditional teaching, the time for students to complete the prescribed experimental operation in this teaching mode was shortened by 4 minutes on average, and the average score of theoretical course test case analysis questions was increased by 1.5 points. In conclusion, the instructional design of the experiment teaching forensic science and biological evidence can effectively improve students' post-competency, and it deserves further exploration and application.

【Objective】 To explore the the potential risks of antiretroviral therapy(ART) drugs on blood safety among blood donors in Shenzhen. 【Methods】 High pressure liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS) was used to measure ART drugs concentrations in the plasma of regular blood donors (negative control group, n=86) and anti-HIV positive individuals (experimental group, n=98, detected from approximately 440 000 blood donors during 2019—2023). The baseline plasma concentrations of ART drugs in the negative control group were clarified, and the impact of ART drugs on blood safety was analyzed. 【Results】 The baseline concentrations of ART drugs were not detected in 86 samples of negative control group. Four positive ART drugs samples were detected in 1∶2 pooled plasma samples of 98 anti-HIV positive blood donors plasma in the resolution test. The ART positive rate of anti-HIV positive donors was 4.08%, with tenofovir, lamivudine and efavirenz detected in three blood donors and lamivudine, lopinavir, ritonavir and zidovudine detected in one blood donor. 【Conclusion】 ART drugs were found among anti-HIV positive blood donors in Shenzhen. Additional research is needed to investigate the motivation of these specific donors, so as to ascertain the groups most susceptible to potential risks, and guarantee blood safety.

Objective:To explore the protective effect and underlying mechanism of hyperbaric oxygen (HBO) on delayed encephalopathy after carbon monoxide poisoning (DEACMP) in mice.Methods:Totally 225 adult male Kunming mice were selected to establish CO poisoning model via intraperitoneal injection carbon monoxide (CO), and were randomly divided into the air control group, CO poisoning group, and HBO group. Each group was further divided into five time points group, that was 1, 3, 7, 14 and 21 d. The mice in the air control group were injected intraperitoneally with the same amount of air, and the HBO group received HBO treatment at the same time every day. DEACMP mice model was screened by behaviors using the open field test, new object recognition test and nesting test, and the content of myelin basic protein (MBP) were assayed. The mouse brain tissue and mitochondrial were prepared and malonialdehyde (MDA) and adenosine triphosphate (ATP) content were measured with ultraviolet spectrophotometer. MBP content in brain tissue and cytochrome C (CytC) content in the mitochondrial were measured by ELISA. The mitochondria membrane potential (MMP) was measured by flow cytometry.Results:Compared with the air control group, the content of carboxyhemoglobin (COHB) in blood increased significantly and the content of MBP in brain tissue decreased significantly in CO poisoning mice. CO poisoning mice showed motor ability and cognitive dysfunction. Compared with the air control group, the contents of MMP, CytC and ATP were significantly decreased ( P<0.01) in the CO poisoning group; while the MDA content was significantly increased ( P<0.01). Compared with the CO poisoning group, mice behaviors were improved significantly ( P<0.05), the content of MBP, MMP, CytC and ATP were increased ( P<0.05), while the MDA content decreased significantly ( P<0.01) in the HBO group. Conclusions:The abnormal mitochondrial function might be closely related to the occurrence and development of DEACMP, and HBO therapy plays an effective role in preventing and treating the DEACMP mice model via the mitochondrial pathway.

Objective:To investigate the risk factors of infection with polymyxin resistant and carbapenemase-resistant Klebsiella pneumoniae（PR-CRKP）. Methods:A total of 170 patients with CRKP infection admitted in the Third Affiliated Hospital of Soochow University from July 2020 to October 2023 were enrolled，including 123 cases of CRKP infection and 47 cases of PR-CRKP infection. The general conditions，exposure of antibacterial drugs 6 months before admission，laboratory test indicators at admission，antibacterial drug use when target bacteria were detected，length of hospital stay and time of invasive procedures in two groups were retrospectively analyzed. The risk factors of PR-CRKP infection were analyzed with univariate and multivariate logistic regression. SPSS 26.0 software was used to analyze the data.Results:Univariate analysis showed that compared with the CRKP group，the average age of patients in PR-CRKP group was older（ Z = -2.186， P = 0.029），the proportion of patients with exposure history to semisynthetic penicillins，carbapenems，polymyxins，and quinolones 6 months before admission was higher（ χ 2= 3.930，5.414，11.939，8.478，all P < 0.05），the proportion of infections diagnosed at admission and blood urea nitrogen levels（ χ 2= 7.268， Z = -2.406， P = 0.007 and 0.016）was higher，the hemoglobin level（ t = 2.641， P = 0.009）was lower，the length of hospital stay was longer，the rates of tracheal intubation，urinary catheter，and deep vein catheterization were higher（ Z = -4.243，-4.660，-5.341，-4.583，all P < 0.001），the duration of carbapenem and polymyxin B use was longer（ Z = -4.757，-7.554，both P < 0.001），the proportion of combined quinolone-resistant Escherichia coli（QREC）and carbapenem-resistant organism（CRO）infections and bloodstream infections，and the rate of admission to intensive care units was higher（ χ 2 = 33.737，42.041，5.426，12.991， P < 0.05 or < 0.01）. Multivariate analysis showed that time to polymyxin B use（ OR = 1.179， 95%CI 1.059-1.312， P = 0.003），combined QREC infection（ OR = 5.357， 95%CI 2.100-13.669， P < 0.001）and combined CRO infection（ OR = 3.302， 95%CI 1.146-9.514， P= 0.027）were independent risk factors for PR-CRKP. Conclusion:Prolonged use of polymyxin B is an independent risk factor for PR-CRKP，and mixed QREC and CRO infection can increase the risk of PR-CRKP.

Objective:To investigate the difference of curative effect between zero-profile bridge-shaped locking cage (ROI-C) and anterior cage combined with titanium plate fixation in the treatment of two-level and three-level cervical spondylotic myelopathy.Methods:A total of 85 patients (43 males and 42 females), aged 52.3±8.0 years (range from 28 to 66 years) with bi- and three-level cervical spondylotic myelopathy who received surgical treatment from June 2017 to October 2019 were retrospectively analyzed. There were 63 cases of two levels and 22 cases of three levels. 45 cases were treated with zero-profile bridge-shaped locking cage ROI-C (ROI-C group), and 40 cases with anterior cage combined with titanium plate fixation (titanium plate group). The main observation indicators include operation time, intraoperative blood loss, cervical Cobb angle, fusion segment Cobb angle, average intervertebral height, pain visual analogue scale (VAS), Japanese Orthopaedic Association (JOA) Score and neck disability index (NDI).Results:All of 85 patients were followed up for 16.9±2.0 months (range 12 to 22 months). The operation time of two-level ROI-C group was 110.37±8.25 min, which was shorter than 139.5±10.54 min of titanium plate group; the intraoperative blood loss was 15.74±8.10 ml, which was less than 23.71±9.70 ml of titanium plate group; the operation time of three-level ROI-C group was 130.00±5.70 min, which was shorter than 162.83±5.59 min of titanium plate group, while the difference in the intraoperative blood loss between the two groups had no statistical significance. One year after operation, Cobb angle of cervical vertebra in double and three-level ROI-C groups were 15.31°±1.55° and 15.20°±0.42°, respectively, which were largerthan 11.23°±2.03° and 9.20°±1.14° before operation; in titanium plate group, they were 15.89°±1.13° and 16.08°±1.88°, which were higher than 11.25°±2.01° and 9.00°±1.60° before operation, and the differences had statistical significance. The differences between the two groups before operation and 1 year after operation had no statistical significance. One year after operation, the VAS scores of double and three-level ROI-C groups were 1.83±0.66 points and 2.60±0.52 points, respectively, which were less than the preoperative 7.49±0.51 points and 7.60±0.52 points; the titanium plate group was 1.79±0.50 points and 2.41±0.51 points, which were less than the preoperative 7.61±0.63 points and 7.42±0.52 points, and the differences had statistical significance. There was no significant difference between the two groups before operation and 1 year after operation. One year after operation, the JOA scores of double and three-level ROI-C groups were 15.00±0.84 points and 14.70±0.95 points, respectively, which were higher than the preoperative 7.20±0.87 points and 6.60±1.27 points; the scores of titanium plate group were 15.29±0.85 points and 14.83±0.58 points, which were higher than the preoperative 6.89±1.03 points and 6.92±0.67 points, and the differences had statistical significance. The differences between the two groups had no statistical significance. The postoperative JOA improvement rate was excellent. Postoperative dysphagia occurred in 1 case (2.22%, 1/45) in ROI-C group and 8 cases (20.00%, 8/40) in titanium plate group, and the difference in the incidence rate between two groups had statistical significance ( χ2=5.32, P=0.02). Conclusion:Both ROI-C and anterior cage combined with titanium plate fixation in the treatment of double and three-level cervical spondylotic myelopathy can achieve good short-term clinical efficacy, with shorter operation time and lower incidence rate of postoperative dysphagia using ROI-C.

Objective: To investigate the clinical effect of percutaneous curved vertebroplasty in the treatment of thoracolum-bar osteoporotic vertebral compression fractures (OVCFs). Methods: All of 85 patients with single thoracolumbar vertebral OVCFs who met the admission criteria from January 2017 to July 2018 were divided into three groups according to the random dig-its table method. They were treated with percutaneous curved vertebroplasty, routine unipedicular PVP and routine bipedicular PVP respectively. There were 25 patients in the percutaneous curved vertebroplasty group, 6 males and 19 females; aged 56-80 years, with an average age of 70.6±9.7 years. Fracture vertebral body distribution: T₁₀ 2 cases, T₁₁ 4 cases, T₁₂ 3 cases, L₁ 9 cases, L₂ 3 cases, L₃ 1 case, L₄ 1 case and L₅ 2 cases. There were 32 patients in the routine unipedicular PVP group, 6 males and 26 fe-males; aged 58-75 years, with an average age of 69.5±9.3 years. Fracture vertebral body distribution: T₁₀ 2 cases, T₁₁ 4 cases, T₁₂ 5 cases, L₁ 11 cases, L₂ 6 cases, L₃ 1 case, L₄ 1 case and L₅ 2 cases. There were 28 patients in the routine bipedicular PVP group, 5 males and 23 females; aged 59-81 years, with an average age of 69.8±8.8 years. Fracture vertebral body distribution: T₁₀ 2 cases, T₁₁ 4 cases, T₁₂ 4 cases, L₁ 10 cases, L₂ 4 cases, L₃ 1 case, L₄ 1 case and L₅ 2 cases. The operation time, injected cement volume, in-traoperative blood loss were recorded and analyzed. Preoperative, postoperative 1 week and 3 months visual analogue scale scores and oswestry disability index were adopted to value the clinical improvements. Preoperative, postoperative 1 week and 3 months relative vertebral height and kyphosis correction, and the cement leakage rate were measured and analyzed. Results: There was no significant difference in the data of gender, age, VAS scores, ODI and distribution of fracture vertebrae among the three groups (P>0.05), and the baseline data was comparable. The average VAS score in the percutaneous curved vertebroplasty group was 2.3±0.5 at 1 week after surgery, that of the routine unipedicular PVP group was 2.4±0.4 and that of the routine bipe-dicular PVP group was 2.4±0.4; the average ODI in the percutaneous curved vertebroplasty group was 19.8%±3.9%, that of the routine unipedicular PVP group was 20.0%±4.1% and that of the routine bipedicular PVP group was 19.9%±3.8%; they were lower than the preoperative data, which were statistically significant (P<0.001). The average relative vertebral height in the percutaneous curved vertebroplasty group was 48.99%±9.23% at 3 months after surgery, that of the routine unipedicular PVP group was 47.11%±10.12% and that of the routine bipedicular PVP group was 46.71%±11.16%; the average kyphosis cor-rection in the percutaneous curved vertebroplasty group was 6.21%±1.94%, that of the routine unipedicular PVP group was 5.22%±2.07% and that of the routine bipedicular PVP group was 5.97%±2.09%; there was 1 cement leakage case in the per-cutaneous curved vertebroplasty group; those of the routine unipedicular PVP group were 4 cases and those of the routine bipe-dicular PVP group were 6 cases; there was no significant difference among the three groups (P>0.05). Operation time 39.10±2.00 min vs 38.70±1.70 min, injected cement volume 3.60±0.11 ml vs 3.50±0.13 ml and blood loss 5.10±0.30 ml vs 5.00±0.40 ml of the percutaneous curved vertebroplasty group and the routine unipedicular PVP group were less than those of the routine bipedicular PVP group, which were statistically significant (P<0.05). Conclution Percutaneous curved vertebroplasty could achieve satisfactory clinical outcomes for OVCFs, with advantages of less operation time, less blood loss, limited X-ray expo-sure, less injected cement volume, and more balanced augmentation for stabilization of the affected vertebrae and total verte-bral column.

Objective To evaluate the efficacy of one stage laparoscopic choledocholithotomy and cholecystectomy vs.ERCP cholecystolithiasis and cholecystectomy (staged) for cholecystolithiasis complicated with choledocholithiasis.Methods From Jan 2015 to Jun 2017,100 consecutive patients underwent randomized one-stage (TCLCBDE + LC) or two-stage (ERCP + subsequent LC) treatment for common bile duct (CBD) stones.Data evaluated were hospital costs,success rate of one-stage versus two-stage management,postoperative morbidity and GIQLI.Results 47 patients underwent ERCP plus LC two-stage treatment while 46 patients underwent one-stage treatment.Overall success rate of primary intervention for CBD stone clearance was 95.7％ and 97.8％ (P =1.0).Postoperative morbidity was 10.6％ vs.6.5％ (P =0.735),and postoperative hospital stay was (16 ± 5) days vs.(11.9 ± 2.9) days (P ＜ 0.01).One-stage laparoscopic transcystic management was the least costly option compared to two-stage management (19 415 ±2 167)yuan vs.(26 767 ±4 387)yuan (P ＜0.01).GIQLI improved faster in one-stage management group.Conclusion Transcystic one-stage management for gall stones with choledocholithiasis results in shorter hospital stay,lower costs and faster recovery than the two-stage management.

BACKGROUND:The phenomenon of osteoinduction by biomaterials has been proven in animal experiments. OBJECTIVE:To investigate whether the ability of a biomaterial to initiate bone formation in ectopic implantation sites improves the performance of osteoinductive biomaterial as a scaffold for tissue-engineered bone. METHODS:We compared ectopic bone formation by combining autologous adipose-derived stromal cells with an osteoinductive and a nonosteoinductive biphasic calcium phosphate ceramic to create a tissue engineering construction in the muscle of dogs. Al implants were implanted in the back muscle of 10 adult dogs for 8 weeks and 12 weeks, including osteoinductive biphasic calcium phosphate ceramic+adipose-derived stromal cells (osteoinductive complex group), osteoinductive biphasic calcium phosphate ceramic (osteoinductive broup), nonosteoinductive biphasic calcium phosphate ceramic+adipose-derived stromal cells (nonosteoinductive complex group), and nonosteoinductive biphasic calcium phosphate ceramic (nonosteoinductive group). Micro-CT analysis and histomorphometry were performed to evaluate and quantify ectopic bone formation. RESULTS AND CONCLUSION:Ectopic bone formation was visible in the osteoinductive complex group and osteoinductive group, and the former group was superior to the latter one in quality of new bone (P<0.05). However, there was no ectopic bone formation in the other two groups. Micro-CT results were consistent with the histomorphological detection. These findings indicate that osteoinductive biphasic calcium phosphate ceramic, as a kind of bone tissue engineering scaffold material, has a better osteogenic capacity, while adipose-derived mesenchymal stem cells serve as seed cells to promote the ectopic bone formation.