Objective To investigate the effectiveness and safety of drug-eluting beads bronchial arterial chemoembolization(DEB-BACE)versus BACE for the treatment of stage Ⅲ-Ⅳ lung squamous cell carcinoma.Methods A total of 104 patients with stage Ⅲ-Ⅳ lung squamous cell carcinoma,who were admitted to the Lishui Municipal Central Hospital of China between January 2013 and August 2021,were enrolled in this study.According to the therapeutic scheme,the patients were divided into DEB-BACE group(n=41)and BACE group(n=63).For patients of DEB-BACE group,Cisplatin at 75 mg/m2 dose and gemcitabine at 1 000 mg/m2 dose(400 mg was used as loaded-drug dose)were injected through a microcatheter,which was followed by embolization with CalliSpheres microspheres loaded with 400 mg of gemcitabine.For patients of BACE group,Cisplatin at 75 mg/m2 and gemcitabatin at 1 000 mg/m2 were injected through a microcatheter,which was followed by arterial embolization with blank microspheres.Three weeks after DEB-BACE or BACE,the patients of both groups were started on intravenous chemotherapy.The primary study endpoint was overall survival(OS).The secondary study endpoints included progression-free survival(PFS),objective response rate(ORR),disease control rate(DCR),adverse reactions,and the remission rate of dyspnea.Results Of the 104 patients,63 received BACE sequential intravenous chemotherapy and 41 received DEB-BACE sequential intravenous chemotherapy.The median OS in DEB-BACE group was 23.0 moths,which was obviously longer than 12.0 months in BACE group(P=0.009).Multivariate Cox regression analysis showed that DEB-BACE treatment was an independent risk factor for OS(HR=0.59,95％ CI:0.38-0.91,Log-rank test P=0.018).Meanwhile,the remission rate of dyspnea in DEB-BACE group was significantly higher than that in BACE group(57.1％ vs 30.6％,P＜0.043).Conclusion Compared with BACE sequential intravenous chemotherapy,DEB-BACE sequential intravenous chemotherapy can significantly prolong the survival time of patients with stage Ⅲ-Ⅳ lung squamous cell carcinoma and significantly improve the symptoms of dyspnea,which has important applications in the treatment of patients with advanced lung squamous cell carcinoma.

Objective To investigate the differences and the immunocompatibility of wild-type (WT), four-gene modified (TKO/hCD55) and six-gene modified (TKO/hCD55/hCD46/hTBM) pig erythrocytes with human serum. Methods The blood samples were collected from 20 volunteers with different blood groups. WT, TKO/hCD55, TKO/hCD55/hCD46/hTBM pig erythrocytes, ABO-compatible (ABO-C) and ABO-incompatible (ABO-I) human erythrocytes were exposed to human serum of different blood groups, respectively. The blood agglutination and antigen-antibody binding levels (IgG, IgM) and complement-dependent cytotoxicity were detected. The immunocompatibility of two types of genetically modified pig erythrocytes with human serum was evaluated. Results No significant blood agglutination was observed in the ABO-C group. The blood agglutination levels in the WT and ABO-I groups were higher than those in the TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups (all P<0.001). The level of erythrocyte lysis in the WT group was higher than those in the ABO-C, TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups. The level of erythrocyte lysis in the ABO-I group was higher than those in the TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups (both P<0.01). The pig erythrocyte binding level with IgM and IgG in the TKO/hCD55 group was lower than those in the WT and ABO-I groups. The pig erythrocyte binding level with IgG and IgM in the TKO/hCD55/hCD46/hTBM group was lower than that in the WT group and pig erythrocyte binding level with IgG was lower than that in the ABO-I group (all P<0.05). Conclusions The immunocompatibility of genetically modified pig erythrocytes is better than that of wild-type pigs and close to that of ABO-C pigs. Humanized pig erythrocytes may be considered as a blood source when blood sources are extremely scarce.

Objective:To study the effects of cathepsin K(CTSK)on the healing process of tooth extraction socket and type H blood vessel angiogenesis in mice.Methods:Ctsk knockout(Ctsk-/-)mice were generated by CRISPR/Cas9 technology,and genotype sequen-cing,general observation,Micro-CT and immunohistochemistry were performed to confirm successful knockout of Ctsk.Then 8 week-old WT and Ctsk-/-mice were used to establish the tooth extraction modle by extracting the left maxillary first molars,and the mice were sac-rificed at the day 7,10,14,21,28 and 35 respectively(n=3)after tooth extraction.Then samples were subjected to stereo microscope and Micro-CT examination.Immunofluorescence staining was used to study the effect of Ctsk knockout on type H blood vessel angiogene-sis.Results:Ctsk knockout did not affect the soft tissue healing of tooth extraction socket,but significantly promoted the bone healing process,and Ctsk deficency significantly enhanced type H blood vessel angiogenesis in the tooth extraction socket.Conclusion:Ctsk knockout can enhance type H vessel angiogenesis,and promote bone healing process of tooth extraction socket in mice.

Objective To investigate the clinical and cytogenetic characteristics of acute myeloid leukemia(AML)patients with NRAS mutations.Methods Newly diagnosed AML patients in our hospital from January 2020 to August 2022 were selected,and their clinical data were retrospectively analyzed.According to NRAS mutations,the patients were divided into NRAS mutation group and NRAS wild group.The clinical characteristics and cytogenetic differences were compared between the two groups.Results A total of 162 newly diagnosed AML patients were included in this study.There were 28 in NRAS mutation group and 134 in NRAS wild group.The peripheral white blood cell count of NRAS mutation group was significantly higher than that of NRAS wild type group(53.10×109/L vs 24.78×109/L,P<0.05).There were no significant differences in the hemoglobin level,platelet count or bone marrow blast cell count between the two groups(P>0.05).The coexisting gene mutation occurred in 25 patients(89.3%,25/28)in NRAS mutation group.The most common coexisting gene mutation was KRAS,with a mutation rate of 28.6%.Compared with NRAS wild group,NRAS mutation group was more likely to obtain KRAS mutations(P<0.05).There was no significant difference in other coexisting mutated genes between the two groups(P>0.05).The proportion of poor prognosis karyotype in the NRAS mutation group was 23.1%,which was significantly higher than that in NRAS wild group(P<0.05).The proportions of favorable and intermediate prognosis karyotypes in NRAS mutation group were 7.7%and 69.2%,respectively,which were not significantly different from those in NRAS wild group(P>0.05).Conclusion The incidence of NRAS mutation is 17.3%in AML patients in this study.Patients with NRAS mutation are more likely to have KRAS mutation and have a higher proportion of poor prognosis karyotype.

Objective To compare the curative effect and safety of drug-eluting beads bronchial artery chemoembolization(DEB-BACE)with those of simple intravenous chemotherapy in treating elderly patients with intermediate to advanced lung cancer.Methods A total of 213 patients aged＞65 years with intermediate to advanced lung cancer,who were admitted to the Lishui Municipal Central Hospital of China between January 2018 and January 2022,were enrolled in this study.According to the therapeutic scheme,the patients were divided into chemotherapy group(n=107)and DEB-BACE group(n=106).After propensity score matching,chemotherapy group and the DEB-BACE group had 42 patients each.The short-term efficacy and the incidence of adverse reactions were compared between the two groups.Survival curve and Log-rank test were used to compare the survival between the two groups.Cox regression analysis was used to analyze the factors influencing the prognostic survival.Results The postoperative one-,3-,and 6-month disease control rate and objective remission rate in DEB-BACE group were better than those in the chemotherapy group.Before propensity score matching,the median progression-free survival(mPFS)time in DEB-BACE group was 7.0 months,which in the chemotherapy group was 6.0 months(P＜0.001).After propensity score matching,the mPFS in DEB-BACE group was 7.0 months,which in the chemotherapy group was 5.0 months(P=0.001).Before propensity score matching,the median overall survival(mOS)time in DEB-BACE group was 23.0 months,which in the chemotherapy group was 20.0 months(P＜0.001).After propensity score matching,the mOS in DEB-BACE group was 24.0 months,which in the chemotherapy group was 18.0 months(P=0.001).Multivariate Cox regression analysis revealed that therapeutic scheme,tumor size,and TNM stage were the influencing factors for OS.In terms of the adverse reactions,the incidences of both the pre-matched and post-matched myelosuppression in DEB-BACE group were lower than those in the chemotherapy group(P＜0.05).Conclusion For the treatment of intermediate to advanced lung cancer in elderly patients,DEB-BACE is superior to simple intravenous chemotherapy in curative efficacy and safety.

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.

Female ; Humans ; Hypoglycemic Agents/therapeutic use* ; Insulin Resistance ; Metformin/therapeutic use* ; Phosphatidate Phosphatase ; Pioglitazone/therapeutic use* ; Polycystic Ovary Syndrome/genetics*

Objective To validate whether the expression of human cluster of differentiation 55 (hCD55) protein in porcine islet cells could inhibit the activation of complement components in human serum. Methods Four adult pigs with WT (wild type), GTKO [α-1, 3-galactosyltransferase (GGTA1) knockout], GTKO/hCD55 and hCD55 genotypes were selected. Islet cells were isolated from WT, GTKO and GTKO/hCD55 pigs, and the purity and insulin secretion function were detected. The expression of hCD55 at the DNA, RNA and protein levels was analyzed by agarose gel electrophoresis, reverse transcription polymerase chain reaction (RT-PCR) and flow cytometry, respectively. Complement-dependent cytotoxicity assay and complement deposition assay were performed under the incubation conditions with fresh human serum. Results The purity of isolated porcine islet cells from three genotype pigs was > 75%, and the glycemic index was > 1. The expression of hCD55 messenger RNA(mRNA) and protein in GTKO/hCD55 porcine islet cells decreased the deposition of human complement component C3c and membrane-attacking complex C5b-9, and reduced the cytotoxicity. Conclusions The expression of hCD55 protein in porcine islet cells could inhibit the activation of human complement and reduce complement-mediated killing effect, indicating that hCD55 protein could exert complement protection effect on porcine islet cells. These findings provide theoretical basis for the application of hCD55 in islet xenotransplantation.

Objective:To explore the changes in the expression of Tau protein and phosphorylated Tau (p-Tau) protein in neurons after spinal cord ischemia-reperfusion injury (SCII).Methods:Ninety-six healthy adult SD rats were randomly divided into a sham operation group ( n=48) and a SCII group ( n=48). Based on the reperfusion time of 3 h, 6 h, 12 h, 24 h, 48 h and 72 h, the SCII group was divided into 6 subgroups ( n=8 per subgroup). Immunohistochemical staining was used to observe the apoptosis of spinal cord neurons in the L 4-L 5 segments and the expression of Tau protein and p-Tau protein. Results:In the sham operation group, the neuron cells were intact, mainly concentrated in the gray matter. Tau protein was seen in a small number of neuron cells, and a small amount of filamentous p-Tau protein in the pernucleus and cytoplasm. There was no significant difference between Tau protein and p-Tau protein expression in neurons at each time point ( P>0.05). In the SCII group, scattered Tau protein was seen in the apoptotic cells while there was a strong positive expression of Tau protein in the non-apoptotic cells. The expression of Tau protein in the SCII group gradually increased after injury, reaching a peak at 48h and plateauing at 72 h, and was significantly different between any 2 time points (except for 72 h) ( P<0.05). In the SCII group, the positive expression of p-Tau protein was observed in the cytoplasm of the apoptotic cells in strips and sheets. It increased rapidly within 6 h but did not change significantly after 6 h, showing no significant difference between any 2 time points afterwards ( P>0.05). There was a statistically significant difference in the expression of Tau protein and p-Tau protein between the SCII group and the sham operation group at each time point ( P<0.05). Conclusion:It is hopeful to reduce the severity of spinal cord injury by regulating the expression of Tau protein and p-Tau protein within 6 to 48 hours after SCII.

Objective: To explore and clarify the correlation between short-term aggressive intrasegmental recurrence (AIR) and functional magnetic resonance imaging after radiofrequency ablation of hepatocellular carcinoma (HCC). Methods: A retrospective analysis of 1 262 patients with HCC who underwent radiofrequency ablation (RFA) in our hospital from January 2012 to June 2018, all patients were confirmed by pathology as HCC, of which 30 patients were found to have AIR during radiographic follow-up within 3 months after surgery, another 35 patients with disease progression who were controlled in a short period of time were randomly selected as the control group. All the enrolled patients underwent dynamic enhanced magnetic resonance imaging (DCE-MRI) and diffusion weighted imaging (DWI) scanning before surgery, and the differences in clinical data, lesion location, and functional magnetic resonance parameters between the two groups were compared, and their correlation with AIR after RFA was analyzed. Chi-square test, t test and Pearson test were used. Results: The lesions in the AIR group were significantly more in the Ⅰand Ⅳsegments than that in the control group (P<0.05), and there was no significant difference in other liver segments (P>0.05). There was no significant difference in the lesion morphology between the two groups (P>0.05), and there was a significant difference in the early enhancement pattern of the lesions (P<0.05), and the early arterial enhancement rate and apparent diffusion coefficient (ADC) value of the AIR group were significantly lower than the control group (P<0.05). Further correlation analysis found that early enhancement of the arterial artery, early arterial enhancement rate, ADC value and lesion location were associated with AIR and were positively correlated, r values were 0.455, 0.633, 0.518, 0.375 and 0.287 (P<0.05). Conclusion The short-term AIR and functional imaging parameters (arterial early enhancement, early arterial enhancement rate, ADC value) and the liver segment (Ⅰ and Ⅳ) were highly correlated with radiofrequency ablation.