Objective:To investigate the safety and therapeutic effect of split liver transplantation (SLT) in clinical application.Methods:This is a retrospective case-series study. The clinical data of 203 consecutive SLT, 79 living donor liver transplantation (LDLT) and 1 298 whole liver transplantation (WLT) performed at the Third Affiliated Hospital of Sun Yat-sen University from July 2014 to July 2023 were retrospectively analyzed. Two hundred and three SLT liver grafts were obtained from 109 donors. One hundred and twenty-seven grafts were generated by in vitro splitting and 76 grafts were generated by in vivo splitting. There were 90 adult recipients and 113 pediatric recipients. According to time, SLT patients were divided into two groups: the early SLT group (40 cases, from July 2014 to December 2017) and the mature SLT technology group (163 cases, from January 2018 to July 2023). The survival of each group was analyzed and the main factors affecting the survival rate of SLT were analyzed. The Kaplan-Meier method and Log-rank test were used for survival analysis.Results:The cumulative survival rates at 1-, 3-, and 5-year were 74.58%, 71.47%, and 71.47% in the early SLT group, and 88.03%, 87.23%, and 87.23% in the mature SLT group, respectively. Survival rates in the mature SLT group were significantly higher than those in the early SLT group ( χ2=5.560, P=0.018). The cumulative survival rates at 1-, 3- and 5-year were 93.41%, 93.41%, 89.95% in the LDLT group and 87.38%, 81.98%, 77.04% in the WLT group, respectively. There was no significant difference among the mature SLT group, the LDLT group and the WLT group ( χ2=4.016, P=0.134). Abdominal hemorrhage, infection, primary liver graft nonfunction,and portal vein thrombosis were the main causes of early postoperative death. Conclusion:SLT can achieve results comparable to those of WLT and LDLT in mature technology liver transplant centers, but it needs to go through a certain time learning curve.

Objective:To investigate the changes of artemin protein expression in diabetic peripheral neuropathy (DPN) and to explore the regulatory effect of human adipose-derived stem cell (ADSC) exosomes on the change of artemin protein expression.Methods:This research was a prospective observational clinical research combined with experimental research. Thirteen DPN patients (9 males and 4 females, aged 32 to 68 years) who were admitted to the First Affiliated Hospital of Air Force Medical University (hereinafter referred to as our hospital) from May 2022 to October 2023 and met the inclusion criteria were selected as DPN group, and 5 non-diabetes patients (4 males and 1 female, aged 29 to 61 years) who were admitted to our hospital in the same period of time and met the inclusion criteria were selected as control group. The toe nerve or sural nerve tissue in the abandoned tissue after debridement or amputation of patients in the two groups was collected. The pathological changes of nerve tissue were observed after hematoxylin-eosin staining; the protein expressions of S100β and artemin in nerve tissue were observed after immunofluorescence staining, and the artemin protein expression was quantified; the protein and mRNA expressions of artemin were detected by Western blotting and real-time fluorescent quantitative reverse transcription polymerase chain reaction, respectively (the sample number in DPN group and control group was 13 and 5, respectively). Twelve male C57BL/6 mice aged 3 to 5 days were collected to isolate Schwann cells, and the cells were divided into conventional culture group cultured routinely, high glucose alone group (cultured with high concentration of glucose solution only), and high glucose+exosome group (cultured with high concentration of glucose solution and extracted human ADSC exosomes). After 24 hours of culture, the cell proliferation activity was detected by cell counting kit 8 ( n=6). After 48 hours of culture, the protein expression of artemin was detected by Western blotting ( n=3). Results:Compared with those in control group, the neural supporting cells decreased and the inflammatory cells increased in the nerve tissue of patients in DPN group, showing typical manifestations of nerve injury. Immunofluorescence staining showed that compared with those in control group, the nuclei was more, and the protein expression of S100β was lower in nerve tissue of patients in DPN group. The protein expression of artemin in nerve tissue of patients in DPN group was 71±31, which was significantly lower than 1 729±62 in control group ( t=76.92, P<0.05). Western blotting detection showed that the protein expression of artemin in nerve tissue of patients in DPN group was 0.74±0.08, which was significantly lower than 0.97±0.06 in control group ( t=5.49, P<0.05). The artemin mRNA expression in nerve tissue of patients in DPN group was significantly lower than that in control group ( t=7.65, P<0.05). After 24 hours of culture, compared with that in conventional culture group, the proliferation activities of Schwann cells in high glucose alone group and high glucose+exosome group were significantly decreased ( P<0.05); compared with that in high glucose alone group, the proliferation activity of Schwann cells in high glucose+exosome group was significantly increased ( P<0.05). After 48 hours of culture, compared with those in conventional culture group, the protein expressions of artemin of Schwann cells in high glucose alone group and high glucose+exosome group were significantly decreased ( P<0.05); compared with that in high glucose alone group, the protein expression of artemin of Schwann cells in high glucose+exosome group was significantly increased ( P<0.05). Conclusions:The protein expression of artemin in nerve tissue of DPN patients is lower than that in normal nerve tissue, which may be related to the reduction of proliferation activity of Schwann cells by high glucose. Human ADSC exosomes may improve the proliferation activity of Schwann cells by increasing artemin protein expression, thereby delaying the progression of DPN.

Objective:To investigate the safety and therapeutic effect of split liver transplantation (SLT) in clinical application.Methods:This is a retrospective case-series study. The clinical data of 203 consecutive SLT, 79 living donor liver transplantation (LDLT) and 1 298 whole liver transplantation (WLT) performed at the Third Affiliated Hospital of Sun Yat-sen University from July 2014 to July 2023 were retrospectively analyzed. Two hundred and three SLT liver grafts were obtained from 109 donors. One hundred and twenty-seven grafts were generated by in vitro splitting and 76 grafts were generated by in vivo splitting. There were 90 adult recipients and 113 pediatric recipients. According to time, SLT patients were divided into two groups: the early SLT group (40 cases, from July 2014 to December 2017) and the mature SLT technology group (163 cases, from January 2018 to July 2023). The survival of each group was analyzed and the main factors affecting the survival rate of SLT were analyzed. The Kaplan-Meier method and Log-rank test were used for survival analysis.Results:The cumulative survival rates at 1-, 3-, and 5-year were 74.58%, 71.47%, and 71.47% in the early SLT group, and 88.03%, 87.23%, and 87.23% in the mature SLT group, respectively. Survival rates in the mature SLT group were significantly higher than those in the early SLT group ( χ2=5.560, P=0.018). The cumulative survival rates at 1-, 3- and 5-year were 93.41%, 93.41%, 89.95% in the LDLT group and 87.38%, 81.98%, 77.04% in the WLT group, respectively. There was no significant difference among the mature SLT group, the LDLT group and the WLT group ( χ2=4.016, P=0.134). Abdominal hemorrhage, infection, primary liver graft nonfunction,and portal vein thrombosis were the main causes of early postoperative death. Conclusion:SLT can achieve results comparable to those of WLT and LDLT in mature technology liver transplant centers, but it needs to go through a certain time learning curve.

Objective To observe the clinical efficacy of Jiangshabanxia nano-paste on nausea and vomiting in end-stage patients and its effect on the quality-of-life (QOL) in cancer patients. Methods 120 end-stage patients with nausea and vomiting symptoms above grade III were randomly divided into observation group and control group. They were treated with Jiangshabanxia nano-paste and placebo paste respectively. The paste patch was changed every 24 hours and used continuously for 7 days. The nausea and vomiting symptom score, the quality-of-life measurement score and KPS score of cancer patients in the two groups were observed to evaluate the curative effect. Results After 7 days of treatment, the symptom scores of nausea and vomiting in the observation group decreased significantly, the KPS score of the observation group increased, and the effective rate was higher than that in the control group. The score of QOL measurement showed that after treatment, the score of main symptom areas and other symptom areas (except external dyspnea, diarrhea and economic difficulties) in the observation group decreased, and the score of overall health area increased. After treatment, the score of main symptom areas and other symptom areas (except external dyspnea, diarrhea and economic difficulties) in the observation group was lower than that in the control group, and the scores of overall health area in the observation group were higher than those in the control group. Conclusion Jiangshabanxia nano-paste has a good clinical efficacy nausea and vomiting in end-stage patients, it also can improve the quality of life end-stage cancer patients.

Diabetic peripheral neuropathy (DPN) is one of the common chronic complications of diabetes, resulting in neuropathy of spinal nerve, cranial nerve, and vegetative nerve. Diabetic distal symmetric multiple neuropathy is the most representative lesion of DPN, including symptoms of bilateral limbs pain, numbness, and paresthesia, etc. DPN is one of the main reasons causing diabetic foot ulcer (DFU). Schwann cells (SCs) are the primary glia cells of the peripheral nervous system, which play very important role in repairing after nerve injury. As the target cells of chronic hyperglycemia, SCs' functions, including the formation of myelin sheath, the secretion of neurotrophic factors, energy supplying for the axon, and the guidance of axon regeneration, etc., are damaged under the action of high glucose. The destroyed functions of SCs can inhibit the repair of damaged nerves and accelerate the progress of DPN. Therefore, if the damage of high glucose to SCs can be effectively reduced, it will provide a new way for the treatment of DPN and DFU and reduce the morbidity of DFU. This review focuses on the function of SCs and its relationship with DPN.

Objective: To analyze the characteristics of individuals positive for SARS-CoV-2 nucleic acid in a centralized isolation site for people entering China in Huzhou City of Zhejiang Province from December 18, 2021 to January 12, 2022, so as to provide insights into the prevention and control of overseas imported COVID-19. Methods: The basic characteristics, nucleic acid detection and epidemiological investigations were collected from individuals positive for SARS-CoV-2 nucleic acid in a centralized isolation site for people entering China from December 18, 2021 to January 12, 2022, and the temporal distribution, population distribution, source of importation, and virus typing were descriptively analyzed. Results : From December 18, 2021 to January 12, 2022, a total of 2 974 individuals in 19 flights were recorded in this centralized isolation site, and 33 cases were tested positive for SARS-CoV-2 nucleic acid, including 21 confirmed cases with common type, 9 confirmed cases with mild type, and 3 cases with asymptomatic infections. There were 11 cases with Omicron infections ( 33.33% ), 5 cases with Delta infections ( 15.15% ), and 17 cases with infection of unidentified types ( 51.52% ). The median interval ( interquartile range ) from the time of entry to the time of a positive test was 4.0 ( 7.0 ) days among all positive cases, 0 ( 4.0 ) day among cases with Omicron infections and 4.5 ( 8.5 ) days among cases with infections of Delta and unidentified types. The positive cases had a mean age of ( 36.97±8.58 ) years, and included 27 men (81.82%). There were 30 cases ( 90.91% ) receiving two and more doses of COVID-19 vaccines, and 7 cases ( 21.21% ) with a previous history of SARS-CoV-2 infections. There were 19 cases ( 57.58% ) from African countries, and 7 of 11 cases with Omicron infections were imported from African countries. Conclusion Omicron infection was predominant among individuals positive for SARS-CoV-2 nucleic acid in this centralized isolation site for people entering China from December 18, 2021 to January 12, 2022, with no severe cases detected, and most positive cases were imported from African countries.

Objective:To evaluate the effect of botulinum neurotoxin A (BoNT/A) on prevention of chronic migraine (CM) in mice and explore the potential mechanism.Methods:Twenty-four male C57BL/6 mice were randomly divided into control group, nitroglycerin (NTG) group, and BoNT/A+NTG group ( n=8). Mice in the latter two groups were intraperitoneally injected with 10 mg/kg NTG on the 1 st, 3 rd, 5 th, 7 th and 9 th d of experiments to establish CM models. Mice in the BoNT/A+NTG group were injected with 0.18 U/100 μL BoNT/A one h before the first injection of NTG. Mice in the control group were injected with the same dose of normal saline. Basal mechanical withdrawal threshold (MWT) and evoked MWT 2 h after NTG in the facial and hindpaw regions on the 1 st, 3 rd, 5 th, 7 th and 9 th d of experiments were evaluated by von Frey filament test. The motor function of mice 2 h after NTG injection was tested by rotarod test on the 1 st, 3 rd, 5 th, 7 th and 9 th d of experiments. On 9 th d of experiments, the mice were sacrified; the calcitonin gene-related peptide (CGRP), synaptosomal-associated protein 25 (SNAP25), glial fibrillary acidic protein (GFAP) and TRP channel protein expressions in the trigeminal ganglia (TG) and trigeminal nucleus caudalis (TNC), and NOD-like receptor protein 3 (NLRP3) inflammatory factor pathway-related protein expressions in TNC were detected by Western blotting; real-time quantitative PCR (RT-qPCR) was used to detect the NLRP3 inflammatory factor pathway-related mRNA expressions in TNC. The CGRP expression in TNC was detected by immunofluorescent staining. Results:(1) As compared with the control group, the NTG group had significantly decreased basal facial MWT on the 7 th and 9 th d of experiments ( P<0.05); as compared with the NTG group, the BoNT/A+NTG group had significantly increased basal facial MWT on the 7 th and 9 th d of experiments ( P<0.05). As compared with the control group, the NTG group had significantly decreased evoked facial MWT on the 5 th and 9 th d of experiments ( P<0.05); as compared with the NTG group, the BoNT/A+NTG group had significantly increased evoked facial MWT on the 5 th and 9 th d of experiments ( P<0.05). As compared with the control group, the NTG group had significantly decreased basal and evoked MWT in the hindpaw regions on the 3 rd, 5 th, 7 th and 9 th d of experiments ( P<0.05); as compared with the NTG group, the BoNT/A+NTG group had significantly increased basal and evoked MWT in the hindpaw regions on the 3 rd, 5 th, 7 th and 9 th d of experiments ( P<0.05). (2) There was no significant difference in running time on rotarod among the three groups ( P>0.05). (3)Western blotting results showed that as compared with those in the control group, the CGRP and SNAP25 protein expressions were significantly increased in TG of the NTG group ( P<0.05); and those in the BoNT/A+NTG group were significantly decreased as compared with those in the NTG group ( P<0.05). As compared with those in the control group, the CGRP and NLRP3 protein expressions were significantly increased in TNC of NTG group ( P<0.05); and those in the BoNT/A+NTG group were significantly decreased as compared with those in the NTG group ( P<0.05). (4)RT-qPCR results showed that as compared with that in the control group, the IL-1β mRNA expression in TNC of the NTG group was significantly increased ( P<0.05), and that in the BoNT/A prevention group was statistically decreased as compared with that in the NTG group ( P<0.05). (5) Immunofluorescent staining results showed that as compared with that in the control group, the CGRP expression in TNC of the NTG group was significantly increased, and that in the BoNT/A+NTG group was significantly decreased as compared with that in the NTG group ( P<0.05). Conclusion:BoNT/A can reduce the SNAP25 expression in TG, reduce the CGRP release in TG and TNC, and prevent CM onset; BoNT/A can regulate NLRP3 level in TNC.

Squalene is widely used in pharmaceutical, nutraceutical, cosmetics and other fields because of its strong antioxidative, antibacterial and anti-tumor activities. In order to produce squalene, a gene ispA encoding farnesyl pyrophosphate synthase was overexpressed in a previously engineered Escherichia coli strain capable of efficiently producing terpenoids, resulting in a chassis strain that efficiently synthesizes triterpenoids. Through phylogenetic analysis, screening, cloning and expression of squalene synthase derived from different prokaryotes, engineered E. coli strains capable of efficiently producing squalene were obtained. Among them, squalene produced by strains harboring squalene synthase derived from Thermosynechococcus elongatus and Synechococcus lividus reached (16.5±1.4) mg/g DCW ((167.1±14.3) mg/L broth) and (12.0±1.9) mg/g DCW ((121.8±19.5) mg/L broth), respectively. Compared with the first-generation strains harboring the human-derived squalene synthase, the squalene synthase derived from T. elongatus and S. lividus remarkably increased the squalene production by 3.3 times and 2.4 times, respectively, making progress toward the cost-effective heterologous production of squalene.
Cloning, Molecular ; Escherichia coli/genetics* ; Humans ; Phylogeny ; Squalene ; Synechococcus

Objective:To investigate the role of NIMA-related kinase-7 (NEK7) in breast cancer (BC) and its potential molecular mechanism.Methods:Quantitative real-time reverse-transcription (RT-qPCR) was used to detect the expression of NEK7 in BC tissue and cell lines. The effect of NEK7 on BC cell proliferation was examined by CCK-8. Proteins interacted with NEK7 were screened in Biological database. The effect of overexpression of NOD-like receptor protein 3 (NLRP3) on BC cell proliferation was evaluated. Western blot was used to detect NLRP3 protein expression, and ELISA was employed to evaluate IL-1β and IL-18 expression level.Result:NEK7 was upregulated in BC tissues and cells, and enforced-expression of NEK7 promoted BC cell proliferation[NEK7 over-expression group: 24 h: (0.33±0.02) , 48 h: (0.59±0.02) , 72 h: (0.76±0.02) ; Blank group: 24 h: (0.30±0.02) , 48 h: (0.45±0.02) , 72 h: (0.62±0.03) ; NEK7 empty vector group: 24 h: (0.32±0.02) , 48 h: (0.46±0.02) , 72 h: (0.63±0.03) ]. There was a positive correlation between NEK7 and NLRP3 ( R=0.13) . Overexpression of NLRP3 increased the proliferation ability of BC cell[NLRP3 over-expression group: 24 h: (0.35±0.02) , 48 h: (0.65±0.02) , 72 h: (0.80±0.03) ; Blank group: 24 h: (0.33±0.02) , 48 h: (0.51±0.02) , 72 h: (0.66±0.03) ; NLRP3 empty vector group: 24 h: (0.34±0.02) , 48 h: (0.52±0.03) , 72 h: (0.66±0.03) ]. NEK7 could positively regulate NLRP3 protein and up-regulate IL-1β (NEK7 over-expression group: 129.96±7.62 pg/ml, Blank group: 19.80±2.42pg/ml, NEK7 empty vector group: 21.30±1.77 pg/ml) and IL-18 (NEK7 over-expression group: 144.08±17.20 pg/ml, Blank group: 16.84±2.34pg/ml, NEK7 empty vector group: 17.64±1.94 pg/ml) expression. Conclusion:The upregulation of NEK7 was involved in the process of BC progression by inducing NLRP3 inflammasome activation, suggesting that NEK7 might be a promising therapeutic target for BC.

Objective: To evaluate the outcome of long-term cognitive function after liver transplantation in children and the role of age factor. Methods: Ninety-five pediatric patients, aged 2 yr and 6 months to 6 yr and 11 months at test, at least 1 yr after liver transplantation, were selected.The children′s cognitive function was assessed using Chinese Wechsler Intelligence Scale for Children.The patients were divided into 2 groups according to the age at transplantation: ≤1 yr group (L1 group, n=65) and > 1 yr group (M1 group, n=22). Results: Compared with the normal value, the scores of verbal comprehension and total intelligence quotient (IQ) were significantly decreased, and the proportion of children who had above-average IQ was decreased 1 yr after liver transplantation, the scores of verbal comprehension were decreased, and the proportion of children who had above-average IQ was decreased in group L1, and the scores of verbal comprehension, visual space and total IQ were significantly decreased, the proportion of children who had above-average IQ was reduced, and the proportion of children who had below-average IQ was increased in group M1(P<0.05). Compared with L1 group, the total IQ score was significantly decreased, the proportion of children who had above-average IQ was reduced, and the proportion of children who had below-average IQ was increased in group M1 (P<0.05). Conclusion The long-term cognitive function of children after liver transplantation is lower than that of normal children, and the long-term cognitive function of children ≤1 yr is better than that of children >1 yr.