Objective To investigate the establishment of a six-gene-edited pig-to-non-human primate kidney xenotransplantation model. Methods The kidney of humanized genetically-edited pig (GTKO/β4GalNT2KO/CMAHKO/hCD55/hCD46/hTBM) was transplanted into a cynomolgus monkey. The survival of the recipient and kidney condition after blood perfusion were observed. The parenchymal echo, blood flow changes, and size of the kidney were monitored on a regular basis. Routine blood test, kidney function test and electrolyte assessment were carried out. Dynamic changes of urine, feces and body mass were monitored. At the end of life, the transplant kidney, heart, liver, spleen, lung, and cecum were collected for pathological examination. Results The recipient died at postoperative 7 d. After blood flow was restored, the kidney was properly perfused, the organ was soft and the color was normal. At the end of the recipient's life, a slight amount of purulent secretion was attached to the ventral side of the kidney, with evident congestion and swelling, showing the appearance of "red kidney". Postoperatively, the echo of renal parenchyma was increased, blood flow was decreased, the cortex was gradually thickened, and a slight amount of effusion surrounded the kidney and abdominal cavity over time. In the recipient, the amount of peripheral red blood cells, hemoglobin, albumin, and platelets was progressively decreased, and serum creatinine level was increased to 308 μmol/L at postoperative 7 d, whereas the K⁺ concentration did not significantly change. Light yellow urine was discharged immediately after surgery, diet and drinking water were resumed within postoperative 3 h, and light yellow and normal-shape stool was discharged. The reddish urine was gradually restored to normal color within postoperative 1 d, which were consistent with the results of the routine urine test. A large amount of brown bloody stool was discharged twice in the morning of 2 d after surgery. Omeprazole was given for acid suppression, and the stool returned to normal at postoperative 4 d. The β₂-microglobulin level was increased to 0.75 mg/L at postoperative 7 d. The body mass was increased by 1.7 kg. Autopsy pathological examination showed interstitial edema and bleeding of the transplant kidney, a large amount of infiltration of lymphocytes and macrophages, infiltration of lymphocytes in the arteriole wall and arterial cavity, accompanied by arteritis changes, lymphocyte infiltration in the cecal stroma and congestion in the spleen tissues. No significant abnormal changes were observed in other organs. Conclusions The humanized genetically-edited pig-to-non-human primate kidney xenotransplantation model is successfully established, and postoperative survival of the recipient is 1 week.

Objective:To investigate the relationship between spinal Mas-related gene receptor C (MrgC) and pathophysiological mechanism of bone cancer pain in mice.Methods:Forty male C3H/HeNCrlVr mice, aged 5-7 weeks, weighing 20-25 g, were selected and divided into 4 groups ( n=10 each) using a random number table method: sham operation group (group S), bone cancer pain group (group P), bone cancer pain + MrgC agonist group (group P-agonist) and bone cancer pain + Mrg C antagonist group (group P-Ab). Preparation of the bone cancer pain model: mouse fibrosarcoma cells (NCTC2472) were injected into the upper tibia of mice in P, P-agonist and P-Ab groups, and the equal volume of D-Hanks balanced salt solution was given instead in S group. Fourteen days later cerebrospinal fluid was intrathecally injected in S and P groups, and MrgC agonist and MrgC antibody were intrathecally injected in P-agonist and P-Ab groups. The mechanical paw withdrawal threshold (MWT) to von Frey stimuli was measured before developing the model (T 0), at 7 days after developing the model (T 1), at 14 days after developing the model (before intrathecal injection, T 2), and at 4, 8 and 12 h after intrathecal injection (T 3-5). Results:Compared with group S, no significant change was found in the MWT at T 0 ( P>0.05), and the MWT was significantly decreased at T 1-T 5 in the other groups ( P<0.05). Compared with group P, the MWT was significantly increased at T 3-T 5 in group P-agonist, and the MWT was significantly decreased at T 3-T 5 in group P-Ab ( P<0.05). Conclusions:Spinal MrgC plays an endogenous protective role in the pathophysiological mechanism of bone cancer pain to a certain extent in mice.

Objective:To evaluate the test-retest reliability of MRI criteria in the 2019 Bosniak classification of cystic renal masses (CRMs) and to analyze the impact of lesions′ property, size and readers′ experience on the test-retest reliability.Methods:From January 2009 to June 2019, 207 patients with 207 CRMs were included in this retrospective study. All of them underwent renal MRI and surgical-pathologic examination. According to Bosniak classification, version 2019, all CRMs were independently classified twice by eight radiologists with different levels of experience. All radiologists were blinded to the pathology of the lesions. By using intraclass correlation coefficient (ICC), test-retest reliability was evaluated for all CRMs and for subgroups with different pathological properties (benign and malignant) and different sizes (≤40 mm and>40 mm). The test-retest reliability of 4 senior readers (≥10 years of experience) and 4 junior readers (<10 years of experience) were evaluated respectively. The comparison of ICC was performed using Z test. Results:The 207 CRMs included 111 benign lesions (83 benign cysts, 28 benign tumors) and 96 malignant tumors. There were 87 lesions with maximum diameter ≤40 mm and 120 with maximum diameter>40 mm. The test-retest reliability (ICC) of each reader for all lesions was 0.776-0.888, the overall ICC was 0.848 (95%CI 0.821-0.872). The ICCs of senior and junior readers were 0.853 (95%CI 0.824-0.880) and 0.843 (95%CI 0.811-0.871) respectively, without significant difference between the two groups ( Z=0.85, P=0.374). The ICC of all readers was 0.827 for benign lesions and 0.654 for malignant lesions, showing significant difference ( Z=2.80, P=0.005). The ICC was 0.770 for lesions ≤40 mm and 0.876 for lesions>40 mm, which was significantly different ( Z=-2.36, P=0.018). For CRM subgroups with different pathological properties and different sizes, there was no significant difference in test-retest reliability between senior and junior readers (all P>0.05). Conclusion:The test-retest reliability of MRI criteria in the 2019 Bosniak classification of CRMs is excellent and unaffected by readers′ experience. The reliabilities are not consistent among CRMs of different pathological properties and different sizes, but all reached the level of good and above.

Clinical data of 85 patients with breast abscess during lactation treated in our hospital from January 2014 to October 2017 were retrospectively analyzed.Among 85 cases,30 patients received traditional abscess incision drainage,35 received needle aspiration,and 20 received negative pressure drainage with small incision.The pus culture findings,average hospitalization expenses,average treatment time,pain score,postoperative breast appearance were evaluated.The pus culture found 24 cases of staphylococcus aureus infection in 85 cases (28％),including 40 cases of Methicillin resistant Staphylococcus aureus (47％).The proportion of central abscess (14/20) in patients with small incision drainage was significantly higher than that of patients with needle aspiration treatment (4/35) (P＜0.05).The average hospitalization expense of the puncture group was significantly lower than that of other two groups (P＜0.01).The average treatment time of abscess incision drainage group was significantly higher than that of other two groups (P＜0.01),and the recovery time was all longer 1 month.The score of pain in the abscess incision drainage group was significantly higher than that in other two groups (P＜0.01),and all patients had moderate to severe pain.The breast shape satisfaction of abscess incision drainage group was significantly lower than that of other two groups (P＜0.01).The analysis indicates that treatment of breast abscess during lactation with needle aspiration and negative pressure drainage has short treatment time,no scars and less pain;and it is suggested that central abscess may be treated early with negative pressure drainage.

Objective To investigate the MRI features of renal oncocytoma(RO). Methods We retrospectively analyzed the data of 26 patients by histologically confirmed with RO in Chinese PLA General Hospital from September 2006 to May 2017 and performed pre-operative MRI and dynamic contrast-enhanced MRI.Lesions were divided into two groups based on the diameter:large group with large than 3 cm(16 cases,16 lesions)and small group with less than 3 cm(10 cases,11 lesions).Features of each lesion were analyzed, including location, pseudocapsules, lipids and other 11 indicators. Tumor imaging features were compared between two groups by univariate and multivariate logistic regression analysis. Results Of 27 RO lesions, 12 were located in the left kidney and 15 in the right kidney. Twenty five lesions appeared exophytic(92.6%,25/27),10 lesions showed exophytic angular interface(37.0%,10/27), 25 lesions showed pseudocapsule(92.6%,25/27),4 lesions appeared lipid(14.8%,4/27),2 lesions showed cystic degeneration or necrosis(7.4%,2/27),1 lesion showed hemorrhage(3.7%,1/27),13 lesions appeared fibrous scar(48.1%,13/27),19 lesions with moderate or intense enhancement in the corticomedullary phase (70.4%, 19/27), 9 lesions with wheel-spoke-like enhancement (33.3%, 9/27), and 2 lesions showed segmental enhancement inversion (7.4%, 2/27). Univariate logistic regression exhibited statistically significant correlation between exophytic angular interface with renal parenchyma,fibrous scar,moderate or intense enhancement in the corticomedullary phase,and wheel-spoke-like enhancement of the tumors both in small and large RO groups with OR value of 0.054, 9.898, 8.400 and 10.000, respectively. In the multivariate logistic regression analysis, exophytic angular interface with renal parenchyma and intense enhancement in the corticomedullary phase were found to be high risk factors with OR value of 0.033 and 15.381,respectively.Conclusions The main manifestation of RO on MRI is that both kidneys can occur, with many exogenesis, pseudocapsules, but less lipids, cystic degeneration, necrosis, hemorrhage and segmental enhanced reversal;smaller lesions(diameter<3 cm)tend to conical interface,while larger lesions (diameter≥3 cm)may have fibrous scars,spoke-shaped enhancement,moderate and significantly enhanced cortical phase characteristics.

Objective To investigate the protective effect and mechanism of environmental enrichment(EE)on radiation induced cognitive dysfunction in mice.Methods A total of 45 female Kunming mice(2-month old)were randomly divided into control group,irradiation group and irradiation plus EE group with 15 in each group.Irradiation group and irradiation plus EE group were treated with a single dose of 4 Gy whole body irradiation,irradiation plus EE group were housed in EE condition for 35 d after irradiation.The object recognition task was used to evaluate the cognitive function of mice.The expression of microglial marker IBA-1 in hippocampus was determined by immunohistochemical staining.The expressions of CD68 and synaptophysin(SYP)proteins in hippocampus were assayed by Western blot.Results Compared with control group,the irradiation group had a low discrimination ratio in object recognition task and had a remarkable low level of SYP expression in hippocampus(t＝3.66,6.84,P＜0.05).In addition,radiation activated microglia in hippocampus by increasing the number of IBA-1-positive cells and enhancing the expression of CD 68(t ＝6.83,5.79,P ＜0.05).Compared with irradiation group,irradiation plus EE group increased the discrimination ratio and the expression of SYP in hippocampus(t＝3.56,4.06,P＜0.05),while the number of IBA-1-positive cells and the expression of CD68 were significantly reduced(t＝7.69,4.59,P＜0.05).Conclusions A single dose of 4 Gy whole body irradiation leads to cognitive dysfunction in mice,while EE could effectively improve the animals′cognitive behavior possibly by inhibiting microglial activation and preventing synapse loss in hippocampus.

Objective To investigate the role of CaM/CaMK-Ⅱ signaling pathways in inflammatory pain in mice.Methods Sixty male C57BL6 mice,weighing 25-27 g,were randomly divided into 3 groups (n =20):control group (group C),complete freunds adjuvant (CFA) group (group F) and KN-93+CFA group (group KF).Saline 50 μl were injected into the right side of the claw in group C.CFA 50 μl were injected into the right claw foot for the preparation of inflammatory pain models in group F.KN-93 45 nmol was injected i.c.v.30 min before CFA injection in group KF.The thermal withdrawal latency (TWL) were measured 30 min before injection,1 h and 4 h after injection.The protein expressions of CaMK-Ⅱ,c-fos and CREB in the spinal cord were measured at above time by Western blot.Results Compared with group C,TWL were lower in groups F and KF 1 h and 4 h after injection (P＜0.05).Compared with groups F,TWL in group KF were higher 1 h and 4 h after injection (P＜0.05).Compared with group C,the protein expressions of p-CaMK-Ⅱ,p-CREB,e-fos and mRNA expression of CaMK-Ⅱ,CREB,c-fos were higher in group F and KF 1 h and 4 h after injection (P＜0.05).Compared with group F,the protein expression of p-CaMK-Ⅱ,p-CREB,c-fos and mRNA expressions of CaMK-Ⅱ,CREB,c-fos in group KF were lower 1 h and 4 h after injection (P＜0.05).Conclusion CaM/CaMK-Ⅱ signaling pathways involved in inflammatory pain in mice.

Objective To evaluate the role of spinal CX3C chemokine receptor 1 (CX3CR1) in inflammatory pain and the relationship with calmodulin (CaM)-calmodulin-dependent protein kinaseⅡ(CaMKⅡ) signaling pathways in mice.Methods Ninety-six pathogen-free healthy male C57BL6 mice,weighing 25-27 g,were divided into 3 groups using a random number table:control group (group C,n=30),inflammatory pain group (group IP,n=36) and CX3CR1 antagonist group (group CA,n=30).Inflammatory pain was induced by injecting complete Freund′s adjuvant (CFA) 50 μl into the plantar surface of right hind paws in IP and CA groups,while the equal volume of normal saline was given instead in group C.In group CA,CX3CR1 antagonist (diluted to 1 μg/5 μl in phosphate buffer solution) was intrathecally injected at 1 h before CFA injection.The thermal paw withdrawal latency (TWL) was measured at 30 min before CFA injection (T0) and 30 min,1 h,2 h and 4 h after CFA injection (T2-4).The animals were then sacrificed,and the spinal cord was removed for determination of the expression of phosphorylated CaMKⅡ (p-CaMKⅡ),phosphorylated cyclic adenosine monophosphate response element-binding protein (p-CREB) and c-fos (by Western blot) and expression of CaMKⅡ,CREB and c-fos mRNA (using real-time polymerase chain reaction).Immunofluorescence was used to determine that p-CAMKⅡ was expressed in microglia.Results Compared with group C,the TWL was significantly shortened at T2-4,and the expression of p-CaMKⅡ,p-CREB and c-fos protein and mRNA was up-regulated at T1-4 in IP and CA groups (P<0.05).Compared with group IP,the TWL was significantly prolonged at T2-4,and the expression of p-CaMKⅡ,p-CREB and c-fos protein and mRNA was down-regulated at T1-4 in group CA (P<0.05).p-CaMKⅡ was co-expressed with the microglial specific biomarker.Conclusion CX3CR1 is involved in the development and maintenance of inflammatory pain through activating CaM-CaMKⅡsignaling pathways in mice.

Objective To investigate the role of calcineurin(CaN)in inflammatory pain in rats.Methods Seventy-five male Harlan-Sprague-Dawley rats,weighting of 200-300 g were randomly divided into 3 groups (n=25): group control (group C),group CFA (complete Freunds adjuvant) (group F) and group CaN+CFA (group NF).100 μl CFA were injected on the right hind claw preparaing for inflammatory pain models in groups F and NF,100 μl saline were injected on the right hind claw in group C.CaN 10 U was intracerebroventricular injected 1 d before CFA injection in group NF.Paw withdrawal thermal latency (PWTL) were measured in 30 min prior to (T0),0.5 h (T1),1 h (T2),2 h (T3) and 4 h (T4) after injection.The expression of CaN and nuclear factor kappa B (NF-κB),IL-1β,TNF-α and IL-10 in spinal cord were measured at each time point.Results The PWTL was significantly shorter at T2-T4 in group F,at T3,T4 in group NF than that at T0and in group C (P<0.05);The PWTL at T2-T4 in group NF was significantly longer than that in group F (P<0.05).CaN protein expression in spinal cord at T1-T4 in group F,at T2-T4 in group NF was significantly lower than that of T0 and in the group C,NF-κB p65 protein expression was significantly higher than that of T0 and in the group C (P<0.05).CaN gene and IL-10 protein content at T2-T4 in groups F and NF were significantly lower than that of group C and at T0,NF-κB gene and IL-1β,TNF-α protein content was significantly higher than that of group C and at T0 (P<0.05).CaN protein and CaN gene expression,IL-10 protein content in spinal cord tissue at T1-T4in group NF was significantly higher than that of group F,NF-κB p65 protein and NF-κB gene expression and contents of IL-1β,TNF-α protein were significantly lower than that of group F (P<0.05).Conclusion CaN adjusts pro-inflammatory and anti-inflammatory cytokines by reducing NF-κB and inhibiting the process of inflammatory pain in rats.

Objective: To study the different clinicopathological characteristics between classic and epithelioid renal angiomyolipoma, and the relationships between clinicopathological characteristics and biological behaviors as basis for clinical treatment. Methods: The clinicopathological and follow-up data for the patients diagnosed with renal angiomyolipoma between 2004 and 2011 were retrospectively reviewed and analyzed. Results: There were 414 cases of renal angiomyolipoma diagnosed over 8 years ago, accounting for 8.1% (414/5 287) of all renal parenchymal tumors. The patients included 122 male and 292 female (male-to-female ratio of 1.0∶2.4), mean age 44.0 years (range 15-74 years). Of these, 195(47.1%) tumors occurred in the left kidney, 212(51.2%) in the right kidney and seven (1.7%) were bilateral. Clinically, some cases presented with hypochondrial pain, hematuria or palpable masses. Histologically, 394(95.2%) were classic angiomyolipoma, 20(4.8%) were epithelioid angiomyolipoma; 54, 23 and 7 cases had hemorrhage, necrosis and cystic degeneration, respectively; and 5, 30 and 14 cases had perirenal fat invasion, atypical cells and polymorphic/giant tumor cell. There was a positive correlation between atypical cells and epithelioid tumor type, respectively. The other clinicopathological parameters did not correlate with histological type. Follow-up data was available in 360 patients, with follow-up period of 3 to 99 months. One case died from other causes. The remaining patients were free of disease. Conclusions Angiomyolipoma is a common renal parenchyma tumor. Clinically, it is usually biologically benign. Histologically, it can be either classic or epithelioid types. The epithelioid type should be differentiated from the classic renal cell carcinoma, Mit family translocation renal tumor and renal hemangioblastoma. Atypical cells, more commonly found in the epithelioid angiomyolipoma, do not affect the clinical prognosis of patients.