Objective:Establishment of a new model of human primary colon cancer transplantation tumor in normal immune mice and to provide a reliable experimental animal model for studying the pathogenesis of colon cancer under normal immunity.Methods:Human colon cancer cells come from colon cancer patients who underwent surgery in the Affiliated Hospital of Jining Medical College in 2017. The mice in the cell control group were inoculated with phosphate buffered solution (PBS) containing colon cancer cells, the microcarrier control group was inoculated with PBS containing microcarrier 6, and the cell-microcarrier complex group was inoculated with the PBS containing colon cancer cell-microcarrier complex. The cells of each group were inoculated under the skin of the right axilla of mice by subcutaneous injection, and the time, size, tumor formation rate and pathological changes under microscope were recorded. The transplanted tumor tissue was immunohistochemically stained with the EnVisiion two-step method, and the tumor formation rate of the transplanted tumor was judged according to the proportion of positive cells in the visual field. The polymerase chain reaction (PCR) method was used to detect the expression of human-specific Alu sequence in mice tumor tissue.Results:After inoculation with tumor cells, the mice in the cell control group and the microcarrier control group did not die and did not form tumors; the mice in the cell-microcarrier complex group had palpable subcutaneous tumors in the right axillary subcutaneously on the 5th to 7th days after inoculation, and tumor formation rate is 67% (10/15), and the tumor volume can reach about 500 mm 3 2 to 3 weeks after vaccination. The immunohistochemistry results showed that CK20, CDX-2 and carcinoembryonic antigen were all positively expressed. The PCR results showed that the expression of human-specific Alu sequence can be detected in the transplanted tumor tissue of tumor-bearing mice. Conclusion:Human primary colon cancer cells used microcarrier 6 as a carrier to form tumors in normal immunized mice, and successfully established a new model of human colon cancer transplantation tumor in normal immune mice.

Objective To establish a new patient-derived xenograft (PDX) model of human liver cancer by inoculating the complex of human primary liver cancer cells and a novel microcarrier (microcarrier 6) into mice with normal immune function. Methods Primary liver cancer cells were isolated and extracted from the fresh human liver cancer tissue of five patients and were then co-cultured with microcarrier 6 to construct a three-dimensional tumor cell culture model in vitro . According to the type of graft, 75 male C57BL/6 mice were divided into cell control group, microcarrier control group, and experimental group (each sample corresponded to three groups, with 15 groups in total and 5 mice in each group). The liver cancer cell-microcarrier complex was implanted into the mice by subcutaneous inoculation, and tumor formation time, tumor formation rate, and histopathological manifestations were observed. The Fisher's exact test was used for comparison of categorical data between two groups. Results As for the liver cancer cells from the five patients, tumor formation was observed in the mice corresponding to three patients. In these three experiments, tumor formation was not observed in the control groups and was only observed in the experimental groups, and 12 of the 15 mice in the experimental groups had successful tumor formation, with a tumor formation rate as high as 80%, which was significantly different from that in the cell control groups and the microcarrier control groups (all P < 0.05). The tumor formation time was 5-7 days; the xenograft tumor grew rapidly, and HE staining showed nested or flaky cells with obvious heteromorphism, with the presence of pathological mitosis; immunohistochemical staining showed positive CK8/18, Hep, and Gpc-3, which was in accordance with the characteristics of human liver cancer cells. Conclusion This experiment successfully establishes a new PDX model of human liver cancer based on the complex of microcarrier 6 and human primary liver cancer cells in mice with normal immunity. This model can be used to better elucidate the mechanism of the development and progression of liver cancer in the body with normal immunity, and besides, it also provides a new animal model with higher value for the precise treatment of liver cancer.

Objective:To analyze the impact of rare bacteria infection on clinical outcome in patients with diabetic foot ulcer(DFU).Methods:A total of 288 cases infected with single strains bacteria were selected. Data were grouped according to the 15 bacteria infection identified. The outcomes of healing, amputation, cardio and cerebrovascular events, and death were collected, and risk factors to the outcome were analyzed.Results:The rare infected bacteria were acinetobacter baumannii, staphylococcus epidermidis, morgan morganella, staphylococcus haemolyticus, streptococcus lactis, streptococcus agalactiae, enterobacter cloacae, and serratia marcescens.There were significant differences in age, albumin, HbA 1C, body mass index, condition of foot ulcer, degree of infection, healing, and minor amputation among these groups. Severe lower extremity arterial disease and age over 70 years were the main risk factors for the healing of ulcers. Wagner grade over 3 and infected with streptococcus lactis were the main risk factors for minor amputation. Severe lower extremity arterial disease, hemoglobin(Hb)≤90 g/L, and albumin(ALB)≤30 g/L were the main risk factors for major amputation. Estimated glomerular filtration rate <60 ml·min -1·(1.73 m 2) -1 and Hb≤90 g/L were the main risk factors for heart failure. Age over 70 years and ALB≤30 g/L were the main risk factors for death(All P<0.05). Conclusion:There exist significant differences in general condition, foot ulcer, and outcome in DFU patients infected with rare bacteria strains.

Objective:Establishment of a new model of human primary colon cancer transplantation tumor in normal immune mice and to provide a reliable experimental animal model for studying the pathogenesis of colon cancer under normal immunity.Methods:Human colon cancer cells come from colon cancer patients who underwent surgery in the Affiliated Hospital of Jining Medical College in 2017. The mice in the cell control group were inoculated with phosphate buffered solution (PBS) containing colon cancer cells, the microcarrier control group was inoculated with PBS containing microcarrier 6, and the cell-microcarrier complex group was inoculated with the PBS containing colon cancer cell-microcarrier complex. The cells of each group were inoculated under the skin of the right axilla of mice by subcutaneous injection, and the time, size, tumor formation rate and pathological changes under microscope were recorded. The transplanted tumor tissue was immunohistochemically stained with the EnVisiion two-step method, and the tumor formation rate of the transplanted tumor was judged according to the proportion of positive cells in the visual field. The polymerase chain reaction (PCR) method was used to detect the expression of human-specific Alu sequence in mice tumor tissue.Results:After inoculation with tumor cells, the mice in the cell control group and the microcarrier control group did not die and did not form tumors; the mice in the cell-microcarrier complex group had palpable subcutaneous tumors in the right axillary subcutaneously on the 5th to 7th days after inoculation, and tumor formation rate is 67% (10/15), and the tumor volume can reach about 500 mm 3 2 to 3 weeks after vaccination. The immunohistochemistry results showed that CK20, CDX-2 and carcinoembryonic antigen were all positively expressed. The PCR results showed that the expression of human-specific Alu sequence can be detected in the transplanted tumor tissue of tumor-bearing mice. Conclusion:Human primary colon cancer cells used microcarrier 6 as a carrier to form tumors in normal immunized mice, and successfully established a new model of human colon cancer transplantation tumor in normal immune mice.

Brain ; pathology ; China ; Humans ; Organ Preservation ; standards ; Tissue Banks ; ethics ; standards

Objective Most common infected bacteria were found to analyze their effects on clinical characteristics and 3-year outcome of patients with diabetic foot ulcer ( DFU ) . Methods Materials of cases with positive bacterial culture were selected from DFU patients. 203 cases were infected with the most common 6 strains of mono-bacteria, and 62 cases were infected with multi-bacteria. Data were grouped according to the most common 6 infected bacteria. The outcomes of healing, recurrence, amputation, cardiac events, cerebrovascular events and death were calculated of 3 years after hospitalization. Clinical characteristics of mono-and multi-infected groups and these 6 mono-bacterial infection groups, and risk factors to outcome were analyzed. Results No significant difference was found in baseline clinical characteristics, cardiac and cerebrovascular events, and death during follow-up between mono-and multi-infected groups. The most common 6 infected bacteria were staphylococcus aureus, pseudomonas aeruginosa, proteus, enterococcus faecalis, escherichia coli and klebsiella pneumoniae. Among these groups, there were no significant differences of baseline clinical characteristics and recurrence, cardiac and cerebrovascular events, and death except for the foot ulcer and foot ulcer related prognosis. In staphylococcus aureus infected group, severe lower extremity arterial disease (8.5％), Wagner grade 3-5 (48.9％), moderate and severe infection rate (34.0％) were significantly lower than other groups, and the healing rate ( 93. 6％) was higher than other groups ( all P<0.05). Severe lower extremity arterial disease, cardiac function grading over 3(NYHA), eGFR<60 ml·min-1· (1.73 m2)-1, duration of DFU over 30 days were the main risk factors for ulcers′healing. Wagner grade over 3 was main risk factor for minor amputation. Severe lower extremity arterial disease, Hb<90g/L were the main risk factors for major amputation. Cardiac function grading over 3 ( NYHA ) was main risk factor for cardiac events, and also for death. ALB<30 g/L was main risk factor for death (all P<0.05). Conclusion DFU patients infected with different strains of bacteria were significantly different in foot ulcer and healing rate, while not in cardiac and cerebrovascular events and death.

Objective Secondary infection with pseudomonas aeruginosa( PA) in diabetic foot ulcer( DFU) was analyzed to investigate the related risk factor, antibiotic resistance, and prognoses of the infection. Methods Pathogen cultures were carried out in 966 DFU patients with their clinical data collected. All of the patients were followed-up for two years to observe the outcomes, including ulcer healing, amputation, recurrence of ulcers, non-fatal cardiovascular events, and death. The antibiotic susceptibility, risk factors and associated outcome of secondary PA infection were analyzed. Results Total incidence of PA infection was 13. 0％ in DFU patients, of which 38. 1％was secondary. The susceptibility rates of secondary infected PA to tobramycin, meropenem, eftazidime, levofloxacin, cefepime, and cefepime were similar to those in primary infected PA. However, the susceptibility rates of secondary infected PA to piperacillin, piperacillin/tazobactam, ciprofloxacin, imipenen, gentamicin, aztreonam, and amikacin decreased by 12％ to 22％ as compared with primary infected PA. The healing rate was much lower in patients with secondary PA infection compared with those with primary PA infection, and the accumulated healing rates at2yearswere44.44％ and70.4％ (P=0.01) respectively. Theriskofulcerhealingfailurewithintwoyears increased by 3 folds in patients with secondary PA infection. After adjusting for age, sex, Wagner grade, infection grade, and duration of DFU, plasma albumin level was an independent risk factor for secondary PA infection in patients with DFU(P=0. 001). Conclusions The antibiotics susceptibility rates of secondary infected PA were lower than those of primary infected PA. Secondary PA infection in DFU was less likely to be healed. Plasma albumin level was a risk factor for secondary PA infection.

Objective To investigate the association of serum C peptide concentration with the severity and the outcome of diabetic foot ulcer (DFU). Methods The clinical data of 257 inpatients with DFU were collected, including fasting and postprandial 2h C peptide levels and C peptide area under curve (AUCCP ). The patients were followed up on the outcomes of ulcers and death. The associations of serum C peptide concentration with the Wagner degree, infection severity, and healing rate were analyzed. Results The medians of fasting and 2h postprandial serum C peptide as well as AUCCP were 1. 37(0. 02 ~ 9. 00) nmol/ L, 3. 22(0. 02 ~ 29. 61) nmol/ L, and 511. 65 (3. 60 ~ 2 691. 30)nmol·min-1 ·L-1 respectively, which were lower than general levels. The time of follow-up in our study was 2. 8 (1. 0 ~ 5. 1) years. By the end of study, the wound of 75. 88% patients was healed, 3. 5%undergone major amputation, and 23. 74% died. After adjusting for relative factors, there were no significant associations of serum fasting and postprandial C peptide levels and AUCCP with Wagner degree and infection severity (P>0. 05). Cox regression analysis showed that the fasting plasma C peptide and hemoglobin were the independent protective factors for the healing of ulcers; old age, male, higher infection degree, and diabetes family history were their independent risk factors ( all P < 0. 05). Conclusions The lower plasma fasting C peptide concentration in patients with DFU is not correlated with Wagner degree and infection severity, but closely related with healing rate.

Objective To compare whether there is a difference in medical cost of intensive care unit(ICU)pa-tients with catheter-associated infection (CAI)between before and after targeted intervention.Methods CAI in ICU patients in 2010(pre-intervention group)and 2013 (post-intervention group)were investigated by retrospective survey,hospitalization cost of two groups of patients before and after intervention was compared.Results The morbidity and mortality in patients with CAI both decreased significantly after intervention,morbidity of healthcare-associated infection(HAI)decreased from 13.47% in 2010 to 4.41 % in 2013,mortality decreased from 10.36% in 2010 to 2.2% in 2103.Total hospitalization cost,blood transfusion cost,and cost of special material before and af-ter the implementation of targeted intervention all significantly different (all P <0.05),the difference of procalcito-nin and antimicrobial agents cost were also significant(all P <0.05).Conclusion Medical cost in ICU patients with CAI decreased after intervention.

Objective Toinvestigatetheoperativeeffectandsafetyofendovascularstentingfor thetreatmentofsymptomaticvertebralarteryostialstenosis.Methods Fortypatientswithsymptomatic vertebral artery ostial stenosis and stenosis rate ≥70% were admitted to the Department of Neurology, China-Japan Friendship Hospital from November 2010 to January 2013 were enrolled retrospectively. All patients received endovascular stenting therapy,15 of them were implanted bare metal stents,and 25 were implanted drug eluting stents. The technical successful rate of operation,perioperation complications,and symptom remission rate of the patients were analyzed. At the same time,stroke and death incident as well as the related ischemic symptoms of the stent vascular feeding area in the follow-up period (13 to 36 months)wereobservedandtherestenosisratewasdocumented.Results Atotalof42stentswereimplanted in 40 patients,and the technical success rate was 100. 0%. The preoperative stenosis rate of vertebral artery ostial stenosis was 75% to 99%(mean 85 ± 7%);the postoperative stenosis rate was 0% to 20%(mean 6 ± 4%). There was no perioperative complication. The clinical symptoms of 19 patients disappeared completely,16 were improved significantly within the follow-up period,and the symptom remission rate was 87. 5%. No stent vascular feeding area related stroke and death occurred. Four patients had transient ischemic attack in posterior circulation,13 had restenosis after procedure (10 of them with bare mental stents and 3 with drug eluting stents). There was significant difference in restenosis rate between the bare mental stents andthedrugelutingstents(10/15vs3/25,P=0.001).Conclusion Endovascularstentingforthe treatment of the severe symptomatic vertebral artery ostial stenosis is a safe and efficient method. Although its restenosis rate is high,but it can improve the symptom of posterior circulation ischemia effectively.