OBJECTIVE To explore suitable storage and transportation conditions for “internet plus drug delivery” under high- temperature conditions. METHODS A survey on high-temperature conditions in summer in Beijing was conducted； a retrospective analysis was conducted on “internet plus drug delivery” orders in our hospital from July 2021 to June 2022， summarizing the proportion and delivery range of drugs under different storage and transportation conditions. Additionally， simulation and validation experiments were performed to investigate optimal drug storage and transportation devices for “internet plus drug delivery” in Beijing under high-temperature conditions in summer. RESULTS The monthly average temperature in Beijing from June to August consistently exceeded 25.0 ℃ between 1991 and 2022. From July 2021 to June 2022， a total of 104 drugs were required to be stored below 25.0 ℃， accounting for 31.23% of the 333 drugs listed in our hospital’s “internet plus drug delivery” catalog in Beijing. These drugs were delivered 1 058 times， accounting for 19.63% of the total deliveries. Simulation and validation experiments demonstrated that the average maximum temperature during the next-day delivery process of “carton + foam box + composite aluminum film pearl cotton + 500 g ice bag×2 + gas column bag” was 9.6 ℃， the average minimum temperature was 2.7 ℃， and all the temperatures remained below 15.0 ℃， which could effectively ensure the quality of drugs. CONCLUSIONS Under the high-temperature conditions in summer in Beijing， the storage and transportation device of “carton + foam box + composite aluminum film pearl cotton + 500 g ice bag×2 + gas column bag” can meet the temperature requirements specified in the drug storage instructions for Beijing intra-city drug delivery.

Objective To evaluate the role of preoperative serological indexes in predicting long-term survival and tumor recurrence of hepatocellular carcinoma (HCC) patients after liver transplantation, aiming to explore its significance in expanding the Milan criteria. Methods Clinical data of 669 recipients undergoing liver transplantation for HCC were retrospectively analyzed. The optimal cut-off value was calculated by the receiver operating characteristic (ROC) curve. The risk factors affecting the overall survival and recurrence-free survival rates of HCC patients after liver transplantation were identified by univariate and multivariate regression analyses. The correlation between preoperative serum liver enzymes and pathological characteristics in HCC patients was analyzed. The predictive values of alpha-fetoprotein (AFP) combined with γ -glutamyl transferase (GGT) and different liver transplant criteria for the survival and recurrence of HCC patients after liver transplantation were compared. Results Exceeded Milan criteria, total tumor diameter (TTD) > 8 cm, AFP > 200 ng/mL and GGT > 84 U/L were the independent risk factors for the overall survival and recurrence-free survival rates of HCC patients after liver transplantation (all P < 0.05). Correlation analysis showed that preoperative serum GGT level was correlated with TTD, number of tumor, venous invasion, microsatellite lesions, capsular invasion, tumor, node, metastasis (TNM) stage, Child-Pugh score and exceeded Milan criteria (all P < 0.05). Milan-AFP-GGT-TTD (M-AGT) criteria were proposed by combining Milan criteria, TTD with serum liver enzyme indexes (AFP and GGT). The 5-year overall survival and recurrence-free survival rates of HCC recipients who met the M-AGT criteria (111 cases of exceeded Milan criteria) were significantly higher than those who met Hangzhou criteria (both P < 0.05), whereas had no significant difference from their counterparts who met the University of California at San Francisco (UCSF) criteria (both P > 0.05). Conclusions Preoperative serological indexes of AFP and GGT could effectively predict the long-term survival and tumor recurrence of HCC patients after liver transplantation. Establishing the M-AGT criteria based on serological indexes contributes to expanding the Milan criteria, which is convenient and feasible.

Mesenchymal stem cell (MSC)-based therapy has emerged as a promising treatment for spinal cord injury (SCI), but improving the neurogenic potential of MSCs remains a challenge. Mixed lineage leukemia 1 (MLL1), an H3K4me3 methyltransferases, plays a critical role in regulating lineage-specific gene expression and influences neurogenesis. In this study, we investigated the role and mechanism of MLL1 in the neurogenesis of stem cells from apical papilla (SCAPs). We examined the expression of neural markers, and the nerve repair and regeneration ability of SCAPs using dynamic changes in neuron-like cells, immunofluorescence staining, and a SCI model. We employed a coimmunoprecipitation (Co-IP) assay, real-time RT-PCR, microarray analysis, and chromatin immunoprecipitation (ChIP) assay to investigate the molecular mechanism. The results showed that MLL1 knock-down increased the expression of neural markers, including neurogenic differentiation factor (NeuroD), neural cell adhesion molecule (NCAM), tyrosine hydroxylase (TH), βIII-tubulin and Nestin, and promoted neuron-like cell formation in SCAPs. In vivo, a transplantation experiment showed that depletion of MLL 1 in SCAPs can restore motor function in a rat SCI model. MLL1 can combine with WD repeat domain 5 (WDR5) and WDR5 inhibit the expression of neural markers in SCAPs. MLL1 regulates Hairy and enhancer of split 1 (HES1) expression by directly binds to HES1 promoters via regulating H3K4me3 methylation by interacting with WDR5. Additionally, HES1 enhances the expression of neural markers in SCAPs. Our findings demonstrate that MLL1 inhibits the neurogenic potential of SCAPs by interacting with WDR5 and repressing HES1. These results provide a potential therapeutic target for promoting the recovery of motor function in SCI patients.
Animals ; Humans ; Rats ; Cell Differentiation ; Intracellular Signaling Peptides and Proteins/therapeutic use* ; Leukemia/metabolism* ; Mesenchymal Stem Cells ; Neurogenesis ; Stem Cells ; Transcription Factor HES-1/metabolism*

Cholangiocarcinoma (CCA) is a category of highly heterogeneous and aggressive malignancy mainly originating from bile duct epithelial cells. The median survival time of untreated CCA patients is approximately 12?24 months, and the effectiveness and durability of surgical resection and neoadjuvant chemotherapy are limited. Results of the next-generation sequencing show that dysregulation of the immune system plays an important role in the pathogenesis of CCA. It has opened up new possibilities for the study of therapies targeting the natural course of aggressive CCA, such as immune checkpoint inhibitors, adoptive cell therapy, and tumor vaccines. Based on the current status of immunotherapy for CCA, the authors review the efficacy and dilemmas of current CCA immunotherapy strategies and look forward to the future treatment prospects of CCA.

Objective To investigate the effect of BIS-guided closed-loop target-controlled infusion on perioperative Th1/Th2 balance in elderly patients undergoing laparoscopic radical gastrectomy under different anesthesia and sedation depths. Methods We applied random number table method to divide 73 elderly patients undergoing elective laparoscopic radical gastrectomy into BIS closed-loop target-controlled infusion group with BIS value of 55(group H, n=36) and BIS value of 45(group L, n=37).Intravenous blood samples were collected immediately before surgery (T1), 2h after surgery (T2), 24h after surgery (T3) and 72h after surgery (T4).IL-2, IL-4, IL-6, IL-10, TNF-α and IFN-γ were determined by flow cytometry microsphere array.We compared operation duration, postoperative PACU stay time and postoperative hospitalization time between two groups. Results Compared with T1, IL-6 and IL-10 concentration in group H at T2, T3 and T4 significantly increased (P < 0.05), IL-4 and TNF-α in group H at T4 were significantly increased (P < 0.05), IL-2 and IFN-γ in group H at T2 and T4 were significantly increased (P < 0.05), the concentration of IL-6 and IL-10 in group L at T2, T3 and T4 were increased (P < 0.05), IL-2 in group L at T4 was decreased (P < 0.05), and IFN-γ/IL-6 in two groups were decreased at T2, T3 and T4(P < 0.05).Compared with group L, IL-6 and IL-10 in group H were significantly increased at T2(P < 0.05), IFN-γ/IL-6 in group H was significantly decreased (P < 0.05), IL-2 and IL-10 in group H were significantly increased at T4(P < 0.05).Operation duration, postoperative PACU stay time and postoperative hospitalization time had no statistical significance between two groups (P > 0.05). Conclusion The anesthesia and sedation depth of BIS-guided closed-loop target-controlled infusion set at 45 is better than 55 in maintaining Th1/Th2 balance in elderly patients undergoing laparoscopic radical gastrectomy, but it has no obvious effect on long-term prognosis.

Objective:To explore the enhancement effect of different concentrations of contrast media on blood vessels and hollow organs in CT enterography.Methods:Sixty patients with CT enterography were enrolled from January to August 2019 at Shanghai Ninth People's Hospital, and were prospectively randomly divided into three groups (group A: 90 ml 400 mg/ml contrast media, group B: 90 ml 350 mg/ml contrast media, and group C: 79 ml 400 mg/ml contrast media). Evaluation parameters included CT value, signal noise ratio (SNR) and contrast noise ratio (CNR) of main abdominal vessels (abdominal aorta, superior and inferior mesenteric atery, jejunal artery, ileum artery, superior and inferior mesenteric vein), jejunum, ileum and inflammatory bowel disease. The overall image quality and direct jejunum ileum artery quality scores were evaluated. One-way ANOVA was used to compare the parameters among the three groups.Results:There was no statistical difference in CT value, SNR and CNR of each measurement index among the three groups in plain scan ( P>0.05). The CT value, SNR and CNR of abdominal aorta, superior mesenteric artery, inferior mesenteric artery, jejunal artery and ileal artery in group A and C were higher than those in group B in arterial phase ( P<0.05), but there was no significant difference between group A and group C ( P>0.05). The CT value, SNR and CNR of superior mesenteric vein, inferior mesenteric vein, normal jejunum, ileum and inflammatory lesions in group A were higher than those in group B and C in venous phase ( P<0.05), but there was no significant difference between group B and group C ( P>0.05). The overall image quality scores of group A, B and C were (3.8±0.7), (3.4±0.6), (3.4±0.6), respectively, with no significant difference ( F=3.075, P=0.054). The direct jejunum ileum artery scores of the three groups were (3.5±0.5), (3.1±0.5), (3.4±0.5), respectively, the difference was statistically significant ( F=3.684, P=0.031). Conclusion:At the same injection rate and scanning parameters, contrast media at 400 mg/ml can provide better vessel and hollow organ enhancement.

Objective: To learn the population,virus status and viral types of hantavirus（HV）hosts in Tiantai County of Zhejiang Province from 2011 to 2018,and to provide evidence for hemorrhagic fever with renal syndrome（HFRS）control. Methods: Rodents in Tiantai County were captured by night trapping method. After the species and age of rodents were identified,the composition of rodent species,dominant species and density of rodents were analyzed. The lungs and blood of rodents were sampled to detect the antigen and antibody of HV by immunofluorescence method. The HV antigen-positive lung samples were detected by RT-PCR with specific primers of HV S fragment,then HV was isolated and identified by inoculating Vero-E6 cells. Results: The average rodent density in Tiantai County from 2011 to 2018 was 4.44%. The rodent density in the field and residential areas were 4.94% and 2.23%,respectively. Ten species of rodents were identified,with Apodemus agrarius dominant in the field and Rattus norvegicus in the residential areas. Sixty-seven lung samples were HV antigen positive（4.13%）,one from Rattus norvegicus and the other sixty-six from Apodemus agrarius. Seventy-nine blood samples were HV antibody positive（4.86%）,all from Apodemus agrarius. Thirty-four HV antigen-positive lung samples were positive（50.75%）after RT-PCR amplification. Twenty-two strains of virus were isolated and all of them were from Apodemus agrarius,including twenty-one strains of Hantaan type（HTN）and one strain of Seoul type（SEO）. Conclusion In Tiantai County,Apodemus agrarius is the main source of HFRS infection;the main epidemic type of HV is HTN and SEO is first found in Apodemus agrarius.

Objective: By investigating the genotype and evolutionary variation of hantavirus (HV) in Tiantai county, a national surveillance site for hemorrhagic fever with renal syndrome (HFRS) was set in Zhejiang province, from 2011 to 2018, to reveal the molecular epidemiological characteristics of hantavirus (HV) in Tiantai. Methods: Total RNA was extracted from ultrasound treated HV antigen- positive rat lung samples in Tiantai from 2011 to 2018. After cDNA was prepared, nested PCR was used to amplify partial sequence of M fragments by using specific primers of HV. The sequences of HV in Tiantai from 2011 to 2018 were compared with other known HV sequences in order to identify the genotype and analyze the evolution and variation of the virus. Results: In 67 HV antigen-positive lung specimens, 31 were positive in nested PCR amplification with type-specific primers, including 30 Hantaan virus (HTNV) positive samples, 1 Seoul virus (SEOV) positive sample, and all the 31 samples were from Apodemus agrarius. The phylogenetic tree based on partial M segment was divided into monophyletic group, 30 strains were distributed in HTNV group and 1 was in SEOV group. The HTNV strain Tiantai T2018-130 was independently in one branch, sharing 84.8%-87.9% homology with other strains both at home and abroad, including 29 strains in HTNV group in Tiantai. The other 29 HTNV strains in Tiantai showed closer relationship. The SEOV strain T2016-31 from Apodemus agrarius showed closer relationship with previous strains of SEOV, Tiantai ZT71, ZT10 and Z37 strains of Wenzhou, Zhejiang province. Conclusions HTNV, the main genotype of HV in Tiantai of Zhejiang province, showed obvious geographic clustering, but the strain T2018-130 was distinct from the others in Tiantai. Meanwhile, by sequence analysis, we confirmed that The SEOV strain T2016-31 existed in in Apodemus agrarius, indicating there was a phenomenon of "spillover" between virus and host in SEOV evolution.

Objective To explore the protective effect and mechanism of hydrogen-rich water (HRW) on ethanol-induced acute gastric injury.Methods The experimental study was conducted.Forty kunming mice were divided into the 4 groups by random number table method:normal control group [0.01 mL/g normal saline (NS)+ 0.03 mL/g NS],HRW group (0.01 mL/g NS +0.03 mL/g HRW),ethanol model group (0.01 mL/g 56°alcoholic drinks +0.03 mL/g NS),HRW treated group (0.01 mL/g 56°alcoholic drinks +0.03 mL/g HRW).Ten mice in each group were administrated twice a day for 7 days.Testing indicators:(1) gastric ulcer index was measured,(2) pathological examination of gastric tissues,(3) activity of serum superoxide dismutase (SOD) and expressions of malondialdehyde (MDA),interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α) were measured,(4) expressions of SOD and MDA in gastric tissues were measured,(5) protein expressions of apoptosis related factors (caspase-3,bax,caspase-9,fas and caspase-8) in gastric tissues were measured by immunohistochemistry,(6) relative expressions of mRNA of apoptosis related factors (caspase-3,bax,caspase-9,fas and caspase-8) in gastric tissues were measured by realtime polymerase chain reaction (RT-PCR).Measurement data with normal distriburion were presented as (x)±s.Comparisons among groups were done using the one-way ANOVA and comparison between groups was done using the LSD-t test.Results (1) Gastric ulcer index was measured:gastric ulcer index of mice in the normal control group,HRW group,ethanol model group and HRW treated group were respectively 0,0,10.40± 1.64 and 3.92 ± 0.23,with statistically significant differences (F=175.050,P＜0.05).There was a statistically significant difference between the ethanol model group and normal control group or HRW treated group (t =19.835,12.352,P＜ 0.05).(2) Pathological examination pathological examination of gastric tissues:① Macropathology of gastric tissues:the surface of the gastric mucosa was normal and smooth in the normal control group and the HRW group,without ulcer,erosion and inflammation.The partial gastric mucosal erosion and ulcer in the ethanol model group was large and very severe.Compared with the ethanol model group,the area of gastric mucosal lesion was reduced in the HRW treated group.② Results of pathological examination of gastric tissues:gastric mucosa in the normal control group and HRW group were integrity.Compared with the normal control group,the partial gastric surface epithelium was degenerate and impaired in the ethanol model group.Compared with the ethanol model group,the gastric mucosal erosion and inflammatory cell infiltration were alleviated in the HRW treated group.(3) Expressions of serum SOD,MDA,IL-6 and TNF-α:expressions of serum SOD,MDA,SOD/MDA and IL-6 were respectively (70±6)U/mL,(7.52±0.23) μmol/L,9.40 ± 1.07,(6.3 ± 1.8) ng/L in the normal control group and (74 ± 4) U/mL,(7.61 ±0.91) μmol/L,9.91 ± 1.55,(5.1 ± 1.6)ng/ L in the HRW group and (101 ± 4) U/mL,(16.95 ± 0.66) μmol/L,5.99±0.17,(19.2±4.9) ng/L in the ethanol model group and (115±5) U/mL,(14.02±0.58) μmol/L,8.23±0.32,(7.1±1.8)ng/L in the HRW treated group,with statistically significant differences among the 4 groups (F=97.405,269.950,16.486,25.663,P＜0.05).The serum TNF-α levels were respectively (53± 14) ng/L,(67± 17) ng/L,(52± 13) ng/L,(58±21) ng/L in the above 4 groups,with no significant difference (F=0.862,P＞0.05).(4) Expressions of SOD and MDA in gastric tissues were measured:expressions of SOD and MDA and SOD/MDA were respectively (93 ± 18) U/mL,(7.90± 1.72) μmol/L,12.48±4.54 in the normal control group and (93±13) U/mL,(6.96± 1.49) μmol/L,13.83±3.40 in the HRW group and (121±31) U/mL,(17.10±4.88) μmoL/L,7.88± 3.70 in the ethanol model group and (143 ± 26) U/mL,(7.31 ± 1.58) μmoL/L,20.00±4.68 in the HRW treated group,with statistically significant differences among the 4 groups (F=5.463,15.051,7.388,P＜ 0.05).(5) The expressions of apoptosis related factors in gastric tissues:the results of immunohistochemistry showed that the levels of caspase-3,bax and fas were repectively 0.065 5± 0.003 7,0.065 7±0.003 0,0.225 4±0.024 3 in the normal control group and 0.065 7±0.002 7,0.064 9±0.003 0,0.246 0±0.022 3 in the HRW group and 0.330 7±0.017 3,0.335 4±0.033 3,0.397 0±0.028 5 in the ethanol model group and 0.096 7±0.003 0,0.084 8±0.001 7,0.375 0±0.035 6 in the HRW treated group,showing statistically significant differences among the 4 groups (F=1 004.222,309.171,48.555,P＜0.05).The levels of caspase-9 and caspase-8 were respectively 0.049 2±0.000 4,0.151 5±0.010 2 in the normal control group and 0.047 9±0.002 0,0.154 00.013 5 in the HRW group and 0.047 0±0.003 7,0.157 2±0.006 2 in the ethanol model group and 0.048 7±0.000 8,0.153 9±0.006 3 in the HRW treated group,with no statistically significant difference among the 4 groups (F=0.998,0.297,P＞0.05).(6) The mRNA expressions of apoptosis related factors in gastric tissues:resutls of RT-PCR showed that relative expressions of mRNA of caspase-3,bax,caspase-9 and fas were respectively 1.00±0.00,1.00±0.00,1.00±0.00,1.00±0.00 in the normal control group and 0.72±0.43,0.66±0.26,1.57±0.31,0.50±0.19 in the HRW group and 3.19±0.87,1.58±0.76,3.04± 1.15,2.84±0.98 in the ethanol model group and 0.49±0.16,0.69±0.25,2.98±0.85,0.53±0.24 in the HRW treated group,showing statistically significant differences among the 4 groups (F=32.106,5.038,9.706,23.387,P＜0.05).The mRNA levels of caspase-8 were respectively 1.00±0.00,1.50±0.60,1.36±0.34,1.32±0.43 in normal control group,HRW group,ethanol model group and HRW treated group,with no significant difference among the 4 groups (F=1.337,P＞0.05).Conclusions Hydrogen-rich water could alleviate ethanolinduced acute gastric injury by antioxidant,anti-inflammatory and anti-apoptosis.Hydrogen-rich water is safe and reliable,without toxic and side effects on the body.

Objective To evaluate the clinical efficacy of limited open reduction combined with percutaneous medial locking plate in treatment of Rüedi-Allg(o)wer type Ⅱ and Ⅲ closed tibial pilon fractures.Methods A retrospective case-control analysis was made on 45 cases of closed tibial pilon fractures treated surgically between June 2008 and December 2015.There were 33 males and 12 females,aged from 26-66 years (mean,44.6 years).All cases were unilateral tibial pilon fractures,among which 18 were on the left while 27 were on the right.Thirty-four cases were combined with fibular fractures.There were 14 cases of type Ⅱ fractures and 31 type Ⅲ fractures according to the Rüedi-Allg(o)wer classification.Using the Tscheme-Gotzen system to evaluate soft tissue injury,two patients were in grade 1,29 patients in grade 2,and five patients in grade 3.On the basis of surgical methods,the cases were divided into Group A,limited open reduction with percutaneous medial locking plate and Group B,conventional open reduction.The operation time,reduction quality,fracture healing time,American Orthopedic Foot and Ankle Society (AOFAS) ankle-hindfoot scale at final follow-up visit and complications were recorded and compared between the two groups.Results The operation time of Group A was shorter than that of Group B (P ＜ 0.05).All patients had been followed up for 12-24 months,among which Group A was 22.5 months and Group B was 20.0 months (P ＞ 0.05).Compared with Group B,Group A was superior in fracture healing time (P ＜ 0.05) and AOFAS ankle-hindfoot scale(P ＜0.05),but was inferior in reduction quality (P ＜ 0.05).Poor wound healing was observed in two cases in each group,yet there was no nonunion in all cases.Conclusion Compared with conventional open reduction,the limited open reduction combined with percutaneous medial locking plate has more advantages in operation time and fracture healing,which can achieve better ankle functions for closed tibial pilon fractures.