AIM: To observe the effect of paricalcitol on intestinal ischemia-reperfusion injury, and to explore the relationship with HMGB1/TLR4/NF-κB signaling pathway. METHODS: Twenty-four SPF-grade healthy adult male C57BL/6J mice were divided into 4 groups (n=6) by random number table: sham operation group (S group), paricalcitol pretreatment+sham operation group (SP group), intestinal ischemia-reperfusion group (IR group) and paricalcitol ischemic preconditioning group (P group). S group and SP group were separated the superior mesenteric artery, IR group and P group were clamped the superior mesenteric artery for 45 minutes and then followed by reperfusion for 2 hours to establish the intestinal ischemia-reperfusion model; SP group and P group were intraperitoneally injected with 0.3 μg/kg paricalcitol 24 hours before surgery, and the other two groups were given equal volume of normal saline. The mice were sacrificed at 2 h after reperfusion, and the intestinal tissue was obtained 5 cm from the terminal ileum. The pathological results were observed under light microscope. The intestinal mucosal injury was scored according to the Chiu's scoring standard. The intestinal tissue diamine oxidase (DAO) and tumor were detected by ELISA. Necrosis factor α (TNF-α) and interleukin 6 (IL-6) content; Western blot was used to detect the expression levels of HMGB1, TLR4 and NF-κB p65 protein in small intestine tissues.RESULTS: Compared with S group and SP group, Chiu's score was increased, the expression of Dao, TNF-α and IL-6 were increased, as well as the expression of HMGB1, TLR4 and NF-κB p65 protein increased significantly in IR group (P< 0.05); Compared with IR group, Chiu's score was decreased, the expression of Dao, TNF-α and IL-6 were decreased, as well as the expression of HMGB1, TLR4 and NF-κB p65 protein decreased significantly in P group (P< 0.05). CONCLUSION: Paricalcitol can alleviate intestinal ischemia-reperfusion injury by inhibiting HMGB1/TLR4/NF-κB signaling pathway and playing an anti-inflammatory role.

Objective:To investigate the whole genome characteristics and mutation of imported 2019-nCoV and trace potential source at the genomics level, the viral genomes from the specimens of a cluster of COVID-19 cases which were imported from a freighter were directly sequenced and determined by high-throughput sequencing technology and bioinformatics analysis.Methods:Throat swab specimens from the 8 confirmed cases of COVID-19 from the same freighter were collected to perform the whole genome sequencing for 2019-nCoV using the targeted genome amplification, combined with Ion S5 next-generation sequencing (NGS) technology. Varieties of online virus analysis platforms was used to classify the viruses and analyze mutation sites in the whole genome. Phylogenetic analysis software was used to construct a phylogenetic tree for the viruses, combined with the epidemiological data of the case, to speculate about the source of the viruses.Results:Eight complete genome sequences of 2019-nCoV was successfully obtained. The complete genomes of the viruses were 29, 822-29, 865 bp in length, with the average sequencing depth of 11 928×-33 588× and the coverage of 99.73%-99.87%; the result of Pangolin classification showed that all the eight 2019-nCoV genomes belong to VOC/Delta (B.1.617.2) lineage. The result of whole genome mutation analysis showed that compared with the Wuhan reference strain, the median number of nucleotide mutations in the eight 2019-nCoV genome sequences was 35 (31 to 38), and the median number of amino acid mutations was 26 (24-28); the mutation sites were distributed in 8 gene coding regions (ORF1a, ORF1b, S, ORF3a, M, ORF7a, ORF8, N). Further analysis revealed that eight 2019-nCoV genomes contained 23 characteristic mutation sites belonging to the Delta (B.1.617.2·AY.2) variants of 2019-nCoV. Since the mutation sites among the eight 2019-nCoV genomes were not completely overlapped, and the epidemiological survey report showed that the freighter stopped at multiple ports and had personnel alternation, it was speculated that the clustered COVID-19 cases might have different origins. Phylogenetic analysis showed that all the eight 2019-nCoV genomes were on the AY.2 sub-lineage of the B. 1.617.2 lineage. This result was consistent with the result of Pangolin classification and mutation analysis.Conclusions:In this study, 8 whole genome sequences of Delta variants were obtained through NGS technology from the clustered COVID-19 cases which were imported from a freighter. The sequencing method and analysis result in this study could provide reference for the 2019-nCoV mutation analysis and tracing the source of the COVID-19 cases for the prevention and control of the COVID-19 epidemic.

Objective:To investigate the morphological characteristics of skull base in children with hemifacial microsomia (HFM) by three-dimensional reconstruction and measurement of skull base.Methods:Three-dimensional spiral CT data of HFM children with unilateral involvement who had not received any treatment from the First Center of Maxillofacial Plastic Surgery, Plastic Surgery Hospital of the Chinese Academy of Medical Sciences from February 2010 to December 2020 were collected. The patients were divided into four groups according to the Pruzansky-Kaban classification standard: Ⅰ, Ⅱa, Ⅱb and Ⅲ. The three-dimensional reconstruction and measurement of the skull base were performed using Mimics 17.0 software. Linear measurements of the skull base included the distance from tuberculum sellae to optic canal (OT), foramen rotundum (FrT), foramen ovale (FT), internal acoustic meatus (IT), hypoglossal canal (HT), and the distance from anterior clinoid process to the lesser wing of the sphenoid bone (ClS) and the petrous ridge of the temporal bone (ClP). The measurement indexes of skull base angle included anterior cranial angle (ACA), middle cranial angle (MCA), posterior cranial angle (PCA), and petrous ridge angle (PRA). SPSS 26.0 software was used for statistical analysis of the measurement result. Paired t-test was used for comparison within each group, and one-way ANOVA was used for comparison between different types. P<0.05 was statistically significant. Results:A total of 40 children with HFM were collected, with 10 cases in each group. The result of skull base angle measurement showed that the MCA of the healthy side of typeⅠHFM was significantly larger than that of the affected side, and the MCA and PRA of the healthy side of type Ⅲ HFM were larger than those of the affected side ( P<0.05). There was no significant difference in the angle of skull base between the healthy side and the affected side of typeⅡa andⅡb HFM ( P>0.05). In terms of linear distance measurement of skull base, FrT and HT on the healthy side of typeⅠHFM were longer than those on the affected side; FrT, IT, HT and ClP on the healthy side of typeⅡa HFM were significantly longer than those on the affected side; IT, HT and ClP on the healthy side of typeⅡb HFM were longer than those on the affected side; and HT on the healthy side of type Ⅲ HFM was longer than those on the affected side ( P<0.05). The difference between FrT and ClP (affected / healthy) of four types of HFM was statistically significant ( P<0.05). Conclusions:The morphology of HFM is different between the affected side and the healthy side. As for the angle of skull base, MCA was obvious smaller in the affected side than that in the healthy side in the type Ⅰ and type Ⅲ HFM. As for the linear measurement, the distance from the hypoglossal canal to the tuberculum sellae on the affected side is shorter than that on the healthy side in each type HFM, indicating that the growth and development of the cranial base on the affected side is affected in a certain degree.

Objective:To investigate the morphological characteristics of skull base in children with hemifacial microsomia (HFM) by three-dimensional reconstruction and measurement of skull base.Methods:Three-dimensional spiral CT data of HFM children with unilateral involvement who had not received any treatment from the First Center of Maxillofacial Plastic Surgery, Plastic Surgery Hospital of the Chinese Academy of Medical Sciences from February 2010 to December 2020 were collected. The patients were divided into four groups according to the Pruzansky-Kaban classification standard: Ⅰ, Ⅱa, Ⅱb and Ⅲ. The three-dimensional reconstruction and measurement of the skull base were performed using Mimics 17.0 software. Linear measurements of the skull base included the distance from tuberculum sellae to optic canal (OT), foramen rotundum (FrT), foramen ovale (FT), internal acoustic meatus (IT), hypoglossal canal (HT), and the distance from anterior clinoid process to the lesser wing of the sphenoid bone (ClS) and the petrous ridge of the temporal bone (ClP). The measurement indexes of skull base angle included anterior cranial angle (ACA), middle cranial angle (MCA), posterior cranial angle (PCA), and petrous ridge angle (PRA). SPSS 26.0 software was used for statistical analysis of the measurement result. Paired t-test was used for comparison within each group, and one-way ANOVA was used for comparison between different types. P<0.05 was statistically significant. Results:A total of 40 children with HFM were collected, with 10 cases in each group. The result of skull base angle measurement showed that the MCA of the healthy side of typeⅠHFM was significantly larger than that of the affected side, and the MCA and PRA of the healthy side of type Ⅲ HFM were larger than those of the affected side ( P<0.05). There was no significant difference in the angle of skull base between the healthy side and the affected side of typeⅡa andⅡb HFM ( P>0.05). In terms of linear distance measurement of skull base, FrT and HT on the healthy side of typeⅠHFM were longer than those on the affected side; FrT, IT, HT and ClP on the healthy side of typeⅡa HFM were significantly longer than those on the affected side; IT, HT and ClP on the healthy side of typeⅡb HFM were longer than those on the affected side; and HT on the healthy side of type Ⅲ HFM was longer than those on the affected side ( P<0.05). The difference between FrT and ClP (affected / healthy) of four types of HFM was statistically significant ( P<0.05). Conclusions:The morphology of HFM is different between the affected side and the healthy side. As for the angle of skull base, MCA was obvious smaller in the affected side than that in the healthy side in the type Ⅰ and type Ⅲ HFM. As for the linear measurement, the distance from the hypoglossal canal to the tuberculum sellae on the affected side is shorter than that on the healthy side in each type HFM, indicating that the growth and development of the cranial base on the affected side is affected in a certain degree.

Objective:To investigate the effect of atorvastatin preconditioning on intestinal ischemia-reperfusion (I/R) injury in mice and the relationship with phosphatidylinositol 3-kinase (PI3K)/serine-threonine kinase (Akt) signaling pathway.Methods:Twenty-four healthy male C57BL/6 mice, aged 6-8 weeks, weighing 18-22 g, were divided into 4 groups ( n=6 each) using a random number table method: sham operation group (S group), I/R group, atorvastatin preconditioning group (A group), atorvastatin plus PI3K inhibitor LY294002 group (AL group). Atorvastatin 10 mg/kg was given by intragastric gavage for 3 consecutive days in A and AL groups, and in addition LY294002 0.3 mg/kg was intraperitoneally injected at 30 min before the last administration of atorvastatin in AL group.Intestinal I/R was produced by occlusion of superior mesenteric artery (SMA) for 45 min followed by 2 h reperfusion in anesthetized mice.The superior mesenteric artery was only isolated but not clamped in S group.The mice were sacrificed at the end of reperfusion, and small intestinal tissues were taken for determination of the pathological changes with a light microscope after HE staining and for determination of wet to dry weight ratio(W/D ratio) and expression of PI3K, phosphorylated Akt (p-Akt), autophagy-related proteins Beclin-1, microtubule-associated protein 1 light chain 3Ⅰ (LC3Ⅰ) and LC3Ⅱ.The intestinal damage was assessed and scored according to Chiu.The ratio of LC3Ⅱ expression to LC3Ⅰ expression (LC3Ⅱ/LC3Ⅰ) was calculated. Results:Compared with S group, Chiu′s scores and W/D ratio were significantly increased, the expression of PI3K and p-Akt was down-regulated, the expression of Beclin-1 was up-regulated, and LC3Ⅱ/LC3Ⅰ ratio was increased in I/R, A and AL groups ( P<0.05). Compared with I/R group, Chiu′s scores and W/D ratio were significantly decreased, the expression of PI3K and p-Akt was up-regulated, the expression of Beclin-1 was down-regulated, and LC3Ⅱ/LC3Ⅰ ratio was decreased in A group ( P<0.05). Compared with A group, Chiu′s scores and W/D ratio were significantly increased, the expression of PI3K and p-Akt was down-regulated, the expression of Beclin-1 was up-regulated, and LC3Ⅱ/LC3Ⅰ ratio was increased in AL group ( P<0.05). Conclusion:Atorvastatin preconditioning can mitigate intestinal I/R injury in mice, and the mechanism is related to activating PI3K/Akt signaling pathway and inhibiting the level of autophagy.

Objective:To evaluate the role of nuclear factor NF-E2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathway in atorvastatin-induced reduction of intestinal ischemia-reperfusion (I/R) injury in mice.Methods:Twenty-four healthy male C57BL/6 mice, aged 6-8 weeks, weighing 18-22 g, were divide into 4 groups ( n=6 each) using a random number table method: sham operation group (S group), intestinal I/R group (I/R group), atorvastatin group (ATV group) and atorvastatin+ Nrf2 inhibitor ML385 group (AM group). Intestinal I/R was produced by occlusion of superior mesenteric artery for 45 min followed by reperfusion.In ATV and AM groups, atorvastatin 10 mg/kg was given by gavage for 3 consecutive days daily at 3 day before establishment of the model, while the equal volume of normal saline was given by gavage in S and I/R groups.Nrf2 inhibitor ML385 30 mg/kg was intraperitoneally injected at 1 h before establishment of the model in group AM.The mice were sacrificed at 2 h of reperfusion, and intestine tissues were obtained for examination of the pathological changes of intestinal tissues (with a light microscope) which were scored according to Chiu, for determination of wet/dry weight ratio (W/D ratio), for detection of the activity of superoxide dismutase (SOD) and content of malondialdehyde (MDA) (by xanthine oxidase method and thiobarbituric acid condensation method) and for determination of the expression of Nrf2 and HO-1 (by Western blot). Results:Compared with S group, the Chiu score, W/D ratio and MDA content were significantly increased, the activity of SOD was decreased, and the expression of Nrf2 and HO-1 was up-regulated in the other 3 groups ( P<0.05). Compared with the group I/R, the Chiu score, W/D ratio and MDA content were significantly decreased, the SOD activity was increased, and the expression of Nrf2 and HO-1 was up-regulated ( P<0.05), and the pathological changes were significantly attenuated in group ATV, and no significant change was found in the parameters mentioned above in AM group ( P>0.05). Compared with the group ATV, the Chiu score, W/D ratio and MDA content were significantly increased, the SOD activity was decreased, the expression of Nrf2 and HO-1 was decreased ( P<0.05), and the pathological changes were significantly aggravated in group AM. Conclusion:The mechanism by which atorvastatin reduces intestinal I/R injury is related to activating Nrf2/HO-1 signaling pathway in mice.

Objective:To evaluate the role of peroxidase proliferator-activated receptor γ (PPARγ)/nuclear factor kappa B (NF-κB) signaling pathway in sodium butyrate-induced reduction of intestinal ischemia-reperfusion (I/R) injury in mice.Methods:Thirty-two SPF-grade healthy adult male C57BL/6J mice, aged 7-9 weeks, weighing 20-25 g, were divided into 4 groups ( n=8 each) using a random number table method: sham operation group (Sham group), intestinal I/R group (IIR group), sodium butyrate group (NaB group) and PPARγ inhibitor GW9662 group (GW9662 group). The model of intestinal I/R was established by occlusion of superior mesenteric artery for 45 min followed by 2-h reperfusion in anesthetized animals.GW9662 2 mg/kg was intraperitoneally injected at 1 h before ischemia in GW9662 group, and sodium butyrate 500 mg/kg was intraperitoneally injected at 30 min before ischemia in NaB and GW9662 groups.Blood samples were obtained via cardiac puncture at 2 h of reperfusion, and the animals were then sacrificed.The intestinal tissues were removed for determination of diamine oxidase (DAO), tumor necrosis factorα (TNF-α) and interleukins 6 (IL-6) concentrations in serum (by enzyme-linked immunosorbent assay) and the expression of PPAR and NF-κB p65 (by Western blot). The damage to intestinal mucous membrane was assessed and scored according to Chiu. Results:Compared with group Sham, the Chiu′s score was significantly increased, levels of DAO, TNF-α and IL-6 in serum and intestinal tissues were increased, expression of PPARγ was down-regulated, and expression of NF-κB p65 was up-regulated in group IIR ( P<0.05). Compared with group IIR, the Chiu′s score, levels of DAO, TNF-α and IL-6 in serum and intestinal tissues were decreased, and expression of PPARγ was up-regulated in group NaB, and expression of NF-κB p65 was up-regulated in NaB and GW9662 groups ( P<0.05). Compared with group NaB, the Chiu′s score, levels of DAO, TNF-α and IL-6 in serum and intestinal tissues were increased, and expression of PPARγ was down-regulated, and expression of NF-κB p65 was up-regulated in group GW9662 ( P<0.05). Conclusion:The mechanism by which sodium butyrate reduces intestinal I/R injury may be related to activating PPARγ/NF-κB signaling pathway and inhibiting inflammatory responses in mice.

AIM: To evaluate the effect of SLC7A11 in dexmedetomidine pretreatment induced reduction of ferroptosis caused by intestinal ischemia-reperfusion (II/R) injury in mice. METHODS: Twenty-four healthy meal SPF C57BL/6J mice, aged 8 weeks, weighing 22-25 g, were randomly divided into Sham operation group (S group), intestinal I/R group (II/R group), dexmedetomidine group (DEX group) and dexmedetomidine plus SLC7A11 inhibitior group (DIKE group), with 6 mice in each group. Intestinal ischemia was induced by occluding the superior mesenteric artery for 45 min followed by 30 min of reperfusion to establish the model of II/R injury. In DEX and DIKE groups, Dexmedetomidine 25 μg/kg was intraperitoneally injected at 30 min before clamping the superior mesenteric artery. The same amount of normal saline was injected in the S group and the II/R group. In DIKE group, SLC7A11 inhibitior Imidazole ketone erastin 50 mg/kg was intraperitoneally injected at 90 min before ischemia. Mice were sacrificed 30 min after reperfusion, and small intestinal tissues in length 5 cm away from the ileocecal valvum were obtained for microscopic examination of pathological changes of intestinal mucosa and for determination of contents of Fe

Objective To investigate the diagnostic value of the ratio of % Micro to % Hypo in the diagnosis of three kinds of common thalassemia and iron‐deficiency anemia (IDA ) .Methods Forty‐nine cases of IDA ,24 cases of mildα‐thalassemia ,24 cases of mildβ‐thalassemia ,24 cases of silent α‐thalassemia and 120 individuals undergoing healthy physical examination were selected as the research subjects and divided into 6 groups:normal group ,IDA group ,mildα‐thalassemia group ,mildβ‐thalassemia group ,silentα‐thalassemia and mild thalassemia group(in duding mild α‐thalassemia group and mild β‐thalassemia group) .The % Micro and %Hypo were deteced in each group and their ratio was calculated .The results were performed the analysis and comparison .Results The % Micro/% Hypo ratio had statistical difference between the mild thalassemia group and IDA group ,between the mild thalasse‐mia and IDA group with the normal group(P<0 .01) .The% Micro/% Hypo ratio had no statistical difference between the silent α‐thalassemia group and normal group ,and between the mild α‐thalassemia group and mildβ‐thalassemia group (P>0 .05) .With the% Micro/% Hypo ratio of 0 .9 as the discriminant value to diagnose mild thalassemia and IDA ,its sensitivity ,specificity and accuracy for diagnosing mild thalassemia were 91 .67% ,91 .89% and 91 .72% respectively ,the sensitivity ,specificity and accuracy in diagno‐sing IDA were 91 .94% ,91 .25% and 91 .18% respectively .Conclusion The ratio of % Micro/% Hypo has good differential diagno‐sis and assisted diagnosis screening value ,but has little value for diagnosing silent α‐thalassemia .

OBJECTIVETo observe the clinical efficacy on cognitive impairment after traumatic brain injury (TBD treated with acupuncture and cognitive training.

METHODSSixty patients were randomized into an observation group and a control group, 30 cases in each one, and 5 cases of them were dropped out due to the earlier discharge. Finally, there were 28 cases in the observation group and 27 cases in the control group. In the control group, the cognitive training and conventional treatment were applied. In the observation group, on the basis of the treatment as the control group, acupuncture was applied to Baihui (GV 20), Fengchi (GB 20), Geshu (BL 17) and Fenglong (ST 40), once a day, for 4 weeks totally. The mini-mental state examination (MMSE) and Montreal cognitive assessment (MoCA) were adopted to evaluate the cognitive function in the patients of post-TBI cognitive impairment.

RESULTS(1) After treatment, the total score in MMSE and the score of each item were increased significantly as compared with those before treatment in the two groups (all P<0. 05). Except for the score of immediate recall, the score in MMSE and the score of each of the other items were increased significantly in the observation group as compared with those in the control group after treatment (all P<0. 05). (2)After treatment, the total score in MoCA and the score of each item were increased significantly as compared with those before treatment in the two groups (all P<. 05). Except for the score of nomenclature item, the total score in MoCA and the score of each of the other items were increased significantly in the observation group as compared with those in the control group after treatment (all P<0. 05).

CONCLUSIONBoth the simple cognitive training and the combined therapy of acupuncture and cognitive training improve MMSE and MoCA scores and relieve the cognitive impairment induced by TBI. But the combined therapy achieves the much better efficacy.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Brain Injuries ; psychology ; therapy ; Cognition ; Cognition Disorders ; psychology ; therapy ; Cognitive Therapy ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome