Objective:To investigate the role and mechanism of mitochondrial DNA N6-methyladenine (6mA) in the hippocampal neurons in vascular cognitive impairment.Methods:(1) In vivo experiments: SPF male rats were randomly divided into sham-operated group and chronic cerebral hypoperfusion (CCH) group ( n=12). CCH models in the CCH group were established by ligating bilateral carotid arteries, while rats in the sham-operated group were only bilaterally dissected without ligation. Exploratory ability was detected by open field test 50 d after modeling, cognitive function was evaluated by novel object recognition test 51-53 d after modeling, and learning and memory abilities were tested by Morris water maze 54-59 d after modeling. And then, rats were sacrificed; ATP concentration and reactive oxygen species (ROS) level in the hippocampal tissues were detected, and neuron apoptosis in the hippocampal CA1 area was detected by TUNEL. (2) In vitro experiments: HT-22 cells were divided into normal control (NC) group, oxygen-glucose deprivation (OGD) group, OGD+siControl group, and OGD+siMETTL4 group. Cells in the NC group were cultured routinely, cells in the OGD group were subjected to low sugar and low oxygen for 12 h, and cells in the OGD+siControl group and OGD+siMETTL4 group were, respectively, transfected with NC-siRNA or METTL4-siRNA after being subjected to low sugar and low oxygen for 12 h. Mitochondria morphology was observed by transmission electron microscopy, ROS was detected by flow cytometry, mitochondria membrane potential was detected by JC-1 fluorescent staining, and mitochondrial complex I and III activity was detected by kit. (3) In vivo and in vitro experiments: METTL4 and DNA 6mA expressions in neuronal mitochondria of rat hippocampal tissues and mitochondria of HT-22 cells were detected by immunofluorescent staining and Western blotting. Results:(1) CCH rats had cognitive impairment: compared with the sham-operated group, CCH group had significantly increased frequency of entering the central area and reduced time in exploring new objects in open field experiment,and significantly decreased frequency of crossing the platform and prolonged escape latency in water maze experiment ( P<0.05). Compared with rats in the sham-operated group, rats in the CCH group had significantly decreased hippocampal ATP content ([18.820±1.177] nmol/L vs. [10.190±0.519] nmol/L) and increased ROS content ([4 488.00±255.70] AU vs. [11 644.00±530.20] AU, P<0.05). TUNEL results showed that the number of apoptotic neurons in the hippocampal CA1 area of CCH group was obviously increased than that in sham-operated group. Immunofluorescent staining results showed that 6mA and METTL4 mainly distributed in the mitochondria of hippocampal neurons in CCH group, and the 6mA and METTL4 expressions were obviously increased compared with those in the sham-operated group. Western blotting results showed that METTL4 expression in the hippocampal mitochondria of CCH group was significantly higher than that in the sham-operated group (1.729±0.168 vs. 1.000±0.000). (2) In vitro experiment: under transmission electron microscope, compared with the NC group, HT-22 cells in the OGD group showed obvious mitochondrial ridge disappearance, membrane rupture and vacuolation. Compared with the OGD group, the OGD+siMETTL4 group had significantly increased ATP production, decreased mtROS production, increased mitochondrial membrane potential, and increased mitochondrial complex I and III activities ( P<0.05). Immunofluorescent staining results showed that the mtDNA 6mA and METTL4 expressions in the OGD group were obviously higher than those in the NC group, and both mainly expressed in the mitochondria; mtDNA 6mA expression in the OGD+siMETTL4 group was obviously lower than that in OGD group. Western blotting results showed that METTL4 expression in the OGD+siMETTL4 group was significantly higher than that in the OGD group (1.578±0.261 vs. 2.970±0.280). Conclusion:Specific high expression of methylase METTL4 in hippocampal neurons of rats with cognitive impairment after CCH promotes the increased mtDNA 6mA expression and leads to mitochondrial energy metabolism disorders and increased ROS, which is speculated to be one of the mechanisms causing vascular cognitive impairment.

Objective To explore the safety and feasibility of a modified superior mesenteric artery(SMA)approach in totally laparoscopic complete mesocolic excision(CME)and D3 lymphadenectomy for right colon cancer.Methods A retrospective analysis was performed on clinical data of 77 cases of totally laparoscopic radical surgery for right colon cancer from April 2021 to April 2023.Before August 2022,42 cases underwent traditional SMA approach(control group,only marking with the ileocolic vascular pedicle as the tail of SMA),while after August 2022,35 cases underwent modified SMA approach(modified group,marking with the Treitz's ligament and ileocolic vascular pedicle as the head and tail of SMA,respectively).There was no statistically significant difference in general information between the two groups(P>0.05).The intraoperative conditions,postoperative recovery,and postoperative complications were compared between the two groups.Results Compared with the control group,the modified group had a shorter surgical time[(147.3±35.8)min vs.(173.4±29.9)min,t =-3.428,P =0.001].There were no statistically significant differences in the number of lymph node dissection,number of positive lymph nodes,drainage volume,exhaust time,postoperative hospital stay,and incidence of complications between the two groups(P>0.05).Conclusion The modified SMA approach for totally laparoscopic radical resection of right colon cancer shortens the surgical time,reduces the difficulty and risk of surgery,and has high safety and feasibility.

Objective:To evaluate the diagnostic efficacy and practicality of the Jaundice color card (JCard) as a screening tool for neonatal jaundice.Methods:Following the standards for reporting of diagnostic accuracy studies (STARD) statement, a multicenter prospective study was conducted in 9 hospitals in China from October 2019 to September 2021. A total of 845 newborns who were admitted to the hospital or outpatient department for liver function testing due to their own diseases. The inclusion criteria were a gestational age of ≥35 weeks, a birth weight of ≥2 000 g, and an age of ≤28 days. The neonate′s parents used the JCard to measure jaundice at the neonate′s cheek. Within 2 hours of the JCard measurement, transcutaneous bilirubin (TcB) was measured with a JH20-1B device and total serum bilirubin (TSB) was detected. The Pearson′s correlation analysis, Bland-Altman plots and the receiver operating characteristic (ROC) curve were used for statistic analysis.Results:Out of the 854 newborns, 445 were male and 409 were female; 46 were born at 35-36 weeks of gestational age and 808 were born at ≥37 weeks of gestational age. Additionally, 432 cases were aged 0-3 days, 236 cases were aged 4-7 days, and 186 cases were aged 8-28 days. The TSB level was (227.4±89.6) μmol/L, with a range of 23.7-717.0 μmol/L. The JCard level was (221.4±77.0) μmol/L and the TcB level was (252.5±76.0) μmol/L. Both the JCard and TcB values showed good correlation ( r=0.77 and 0.80, respectively) and agreements (96.0% (820/854) and 95.2% (813/854) of samples fell within the 95% limits of agreement, respectively) with TSB. The JCard value of 12 had a sensitivity of 0.93 and specificity of 0.75 for identifying a TSB ≥205.2?μmol/L, and a sensitivity of 1.00 and specificity of 0.35 for identifying a TSB ≥342.0?μmol/L. The TcB value of 205.2?μmol/L had a sensitivity of 0.97 and specificity of 0.60 for identifying TSB levels of 205.2 μmol/L, and a sensitivity of 1.00 and specificity of 0.26 for identifying TSB levels of 342.0 μmol/L. The areas under the ROC curve (AUC) of JCard for identifying TSB levels of 153.9, 205.2, 256.5, and 342.0 μmol/L were 0.96, 0.92, 0.83, and 0.83, respectively. The AUC of TcB were 0.94, 0.91, 0.86, and 0.87, respectively. There were both no significant differences between the AUC of JCard and TcB in identifying TSB levels of 153.9 and 205.2 μmol/L (both P>0.05). However, the AUC of JCard were both lower than those of TcB in identifying TSB levels of 256.5 and 342.0 μmol/L (both P<0.05). Conclusions:JCard can be used to classify different levels of bilirubin, but its diagnostic efficacy decreases with increasing bilirubin levels. When TSB level are ≤205.2 μmol/L, its diagnostic efficacy is equivalent to that of the JH20-1B. To prevent the misdiagnosis of severe jaundice, it is recommended that parents use a low JCard score, such as 12, to identify severe hyperbilirubinemia (TSB ≥342.0 μmol/L).

Objective To collect information from the traditional Chinese medicine(TCM)four diagnostic information of patients with coronary heart disease(CHD)and to observe the characteristics of their Chinese medicine syndrome factor at different ages,to provide an objective basis for the Chinese medicine treatment of patients with CHD at different ages.Methods The TCM Four Diagnostic Information Collection Scale for CHD developed by the group was used to collect the four diagnostic information of patients with coronary heart disease,and the syndrome elements were extracted according to the syndrome differentiation standard to observe the distribution of syndrome elements at different ages.Results CHD is located in the heart and accompanied by liver,kidney,stomach,lung and other organs.Qi deficiency is the most common syndrome factor,followed by phlegm turbidity,yin deficiency,blood stasis and qi stagnation.Among the syndrome elements of disease location,the heart and liver were the most common in the young group,and the heart and kidney were more common in the middle-aged group and the elderly group.Among the syndrome elements of disease nature,qi deficiency was more common in the young group,phlegm turbidity and qi stagnation were more common in the empirical group,and qi deficiency and phlegm turbidity were the most frequent.In the middle-aged group,qi deficiency was more common in deficiency syndrome,phlegm turbidity and blood stasis were more common in excess syndrome,and the frequency of qi deficiency and phlegm turbidity and blood stasis was the highest.In the elderly group,qi deficiency and yin deficiency were more common in deficiency syndrome,phlegm turbidity and blood stasis were more common in excess syndrome,and qi and yin deficiency combined with phlegm turbidity and blood stasis had the highest frequency.Conclusion The combination of syndrome elements in patients with CHD in different age groups has its own characteristics.The young,middle and elderly groups are all visible in the mixture of deficiency and excess,while the young group is the most common with qi deficiency and phlegm turbidity,the middle-aged group is the most common with qi deficiency and phlegm turbidity and blood stasis,and the elderly group is the most common with qi and yin deficiency and phlegm turbidity and blood stasis.With the increase of age,the complexity of the combination of syndrome elements is higher.

ObjectiveTo observe the effect of the Fangfeng Tongshengsan on post-chemoembolization syndrome with primary liver cancer or postoperative liver metastases of colorectal cancer. MethodSeventy-two patients suffered from post-chemoembolization syndrome after transcatheter hepatic arterial chemoembolization were randomly divided into 2 groups， including a Fangfeng Tongshengsan group and a control group， with 36 patients in each group. The patients in Fangfeng Tongshengsan group orally took the decoction for consecutive 7 d. The patients in the control group were physically cooled down with alcohol rub bath and ice pack for consecutive 7 d. Furthermore， the difference of fever， Karnofsky performance status （KPS）， pain in the liver region， nausea vomiting， constipation， and liver function between these two groups were observed. ResultCompared with the control group， Fangfeng Tongshengsan significantly relieved fever， reduced the body temperature （P<0.05）， and shortened the duration of fever （P<0.05）， indicating that Fangfeng Tongshengsan remarkably improved the KPS （P<0.05）. Meanwhile， Fangfeng Tongshengsan obviously alleviated nausea， vomiting， and constipation status and shortened the duration time compared with the control group （P<0.05）. In addition， the parameters of liver function including alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， γ-glutamyl transpeptidase （GGT）， and total bilirubin （TBIL） were significantly decreased in the Fangfeng Tongshengsan group （P<0.05）， which indicated that Fangfeng Tongshengsan alleviated liver dysfunction of patients with post-chemoembolization syndrome. ConclusionFangfeng Tongshengsan can be used to treat post-chemoembolization syndrome with primary liver cancer and postoperative liver metastases of colorectal cancer.

ObjectiveTo explore the mechanism of Huanglian Ejiaotang in intervening in insomnia based on 5-hydroxytryptamine （5-HT） system and gut microbiota. MethodFifty-five SPF-grade SD rats were randomly divided into normal group， model group， low-， medium-， and high-dose Huanglian Ejiaotang groups （1.925， 3.85， and 7.7 g·kg^-1）， and Estazolam group （0.1 mg·kg^-1）. Except for those in the normal group， the rats in the other five groups were subjected to sleep deprivation on a narrow platform for 12 hours daily for 21 consecutive days. After 14 days of drug intervention， the sleep， exploratory behavior， and depressive-like behavior of the rats were assessed using the pentobarbital sodium sleep synergistic test， the open field test， and the sugar preference test， respectively. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of 5-HT， 5-hydroxyindoleacetic acid （5-HIAA）， tryptophan hydroxylase （TPH）， and monoamine oxidase-A （MAO-A）. Real-time polymerase chain reaction （Real-time PCR） was used to measure the mRNA expression of the 5-HT1A receptor （5-HT1AR） and 5-HT2A receptor （5-HT2AR）. Differences in gut microbiota among the groups were assessed using 16S rRNA sequencing， and the correlation between the 5-HT system and microbiota was revealed using redundancy analysis. ResultCompared with the normal group， the model group showed a prolonged sleep latency （P<0.05）， reduced sleep maintenance （P<0.01）， decreased central area activity time in the open field （P<0.01）， and reduced sugar preference rate （P<0.05）. Moreover， the model group also showed decreased levels of 5-HT， 5-HIAA， TPH， and MAO-A （P<0.01）， decreased 5-HIAA/5-HT ratio （P<0.01）， downregulated mRNA expression of 5-HT1AR （P<0.01）， and upregulated mRNA expression of 5-HT2AR （P<0.05）. The proportion of Firmicutes decreased， while that of Bacteroidetes increased， leading to a decreased Firmicutes/Bacteroidetes （F/B） ratio （P<0.05）. Compared with the model group， the high-dose Huanglian Ejiaotang group exhibited a shortened sleep latency （P<0.01）， and increased sleep maintenance （P<0.01）. The low-dose Huanglian Ejiaotang group showed increased central area activity time （P<0.01） and an increased sugar preference rate （P<0.05）. The high-dose Huanglian Ejiaotang group exhibited increased levels of 5-HT， 5-HIAA， TPH， and MAO-A （P<0.01）， increased 5-HIAA/5-HT ratio （P<0.05）， upregulated mRNA expression of 5-HT1AR （P<0.01）， and downregulated mRNA expression of 5-HT2AR （P<0.05）. The low-dose Huanglian Ejiaotang group displayed an increased proportion of Firmicutes and a decreased proportion of Bacteroidetes， resulting in an increased F/B ratio. At the phylum level， 5-HT， 5-HIAA， and MAO-A were positively correlated with Firmicutes and negatively correlated with Bacteroidetes. At the genus level， 5-HT， 5-HIAA， TPH， and MAO-A were negatively correlated with Prevotella and Lactobacillus and positively correlated with Blautia and Bacteroides. ConclusionHuanglian Ejiaotang can improve sleep deprivation-induced insomnia and depressive-like behavior by regulating the activity of the 5-HT system and the composition of gut microbiota.

Objective:To establish and internally validate a nomogram model for predicting complicated acute appendicitis (CA).Methods:The clinical data from 663 acute appendicitis patients from the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine from October 2015 to October 2022 were retrospectively analyzed. There were 411 males and 252 females, aged ( M (IQR)) 41 (22) years (range: 18 to 84 years). There were 516 cases of CA and 147 cases of uncomplicated acute appendicitis. The minimum absolute contraction and selection operator regression model was used to screen the potential relative factors of CA, and the screened factors were included in the Logistic regression model for multivariate analysis. Software R was used to establish a preoperative CA nomogram prediction model, the receiver operating characteristic curve of the model was drawn, and the value of area under the curve (AUC) was compared to evaluate its identification ability, and the Bootstrap method was used for internal verification. Results:The elderly (age≥60 years) ( OR=2.428, 95% CI: 1.295 to 4.549), abdominal pain time (every rise of 1 hour) ( OR=1.089, 95% CI: 1.072 to 1.107), high fever (body temperature≥39 ℃) ( OR=1.122, 95% CI: 1.078 to 1.168), total bilirubin (every rise of 1 μmol/L) ( OR=2.629, 95% CI: 1.227 to 5.635) were independent relative factors of CA (all P<0.05). The AUC of this model was 0.935 (95% CI: 0.915 to 0.956). After internal verification using the Bootstrap method, the model still had a high discrimination ability (AUC=0.933), and the predicted CA curve was still in good agreement with the actual clinical CA curve. Conclusion:The clinical prediction model based on the elderly (age≥60 years), prolonged abdominal pain time, high fever (body temperature≥39 ℃), and increased total bilirubin can help clinicians effectively identify CA.

Objective:To establish and internally validate a nomogram model for predicting complicated acute appendicitis (CA).Methods:The clinical data from 663 acute appendicitis patients from the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine from October 2015 to October 2022 were retrospectively analyzed. There were 411 males and 252 females, aged ( M (IQR)) 41 (22) years (range: 18 to 84 years). There were 516 cases of CA and 147 cases of uncomplicated acute appendicitis. The minimum absolute contraction and selection operator regression model was used to screen the potential relative factors of CA, and the screened factors were included in the Logistic regression model for multivariate analysis. Software R was used to establish a preoperative CA nomogram prediction model, the receiver operating characteristic curve of the model was drawn, and the value of area under the curve (AUC) was compared to evaluate its identification ability, and the Bootstrap method was used for internal verification. Results:The elderly (age≥60 years) ( OR=2.428, 95% CI: 1.295 to 4.549), abdominal pain time (every rise of 1 hour) ( OR=1.089, 95% CI: 1.072 to 1.107), high fever (body temperature≥39 ℃) ( OR=1.122, 95% CI: 1.078 to 1.168), total bilirubin (every rise of 1 μmol/L) ( OR=2.629, 95% CI: 1.227 to 5.635) were independent relative factors of CA (all P<0.05). The AUC of this model was 0.935 (95% CI: 0.915 to 0.956). After internal verification using the Bootstrap method, the model still had a high discrimination ability (AUC=0.933), and the predicted CA curve was still in good agreement with the actual clinical CA curve. Conclusion:The clinical prediction model based on the elderly (age≥60 years), prolonged abdominal pain time, high fever (body temperature≥39 ℃), and increased total bilirubin can help clinicians effectively identify CA.

Objective:To investigate the protective effect of carbon monoxide releasing molecule 2 (CORM-2) on intestinal barrier by regulating the differentiation of T lymphocytes in rats with hemorrhagic shock.Methods:Healthy male Sprague-Dawley rats ( n=56) were randomly (random number) divided into the sham operation group, shock group, dimethyl sulfoxide (DMSO) control group, inactivated carbon monoxide release molecule-2 (iCORM-2) pretreatment group and three pretreatment CORM-2 with the doses of 2, 4 and 6 mg/kg separately. The hemorrhagic shock was induced with the use of a modified Wiggers model. Rats in the CORM-2 group and iCORM-2 group were intraperitoneally injected with CORM-2 with the doses of 2, 4 and 6 mg/kg and 6 mg/kg iCORM-2 instantly before shock induction. Rats in the DMSO group received intraperitoneal administration of 2% DMSO with the same volume of iCORM-2. Rats in the shock group and sham operation group were not pretreated before inducing shock. Mean arterial pressure of each rat was recorded at different phases after catheterization or shock. Twenty-three hours after shock induction, the permeability of intestinal barrier was measured by FITC-dextran flux, and then ileum tissues were harvested to observe histopathologic changes. Immunohistochemistry was used to detect the expression of transcription factors T-bet and Foxp3 of intestinal mucosa in rats, and the expression of interferon-γ (IFN-γ), interleukin-10 (IL-10) and transforming growth factor-β (TGF-β) in intestinal mucosa was measured by Western blot. One-way analysis of variance or Kruskal Wallis rank sum test was used to compare the difference among groups for normal or non-normal distributed data. Results:Compared with the sham operation group, serum concentrations of FITC-dextran were significantly increased in the shock group, DMSO group and iCORM-2 group (all P<0.05). The concentrations of FITC-dextran in serum of three CORM-2 pretreatment groups pretreatment were significantly decreased compared with other groups undergoing shock (all P<0.05). Rats in the shock group, DMSO group and iCORM-2 group showed severe ileum injury. CORM-2 intervention resulted in alleviation of intestinal mucosal injury in rats with shock, and rats in groups pretreatment CORM-2 at the doses of 4 and 6 mg/kg exhibited integrity of anatomic ileac structure. Compared with the sham operation group, T-bet levels of intestinal intraepithelial lymphocytes were increased in shock group and DMSO group (all P<0.05). Compared with the shock group, levels of T-bet were decreased in intestinal epithelium of three groups pretreatment with CORM-2 at the doses of 2, 4 and 6 mg/kg (all P<0.05). Foxp3 levels in intestinal intraepithelial lymphocytes of the iCORM-2 group and two groups pretreatment with CORM-2 at the doses of 4 and 6 mg/kg were increased compared with the shock group and DMSO group (all P<0.05), but there was no significant difference among the shock group, DMSO group and group pretreatment with CORM-2 at 6 mg/kg (all P>0.05). The shock group showed increased expression of IFN-γ (all P<0.05), but unchangeable expression of IL-10 and TGF-β (all P>0.05) compared with the sham operation group. Compared with the shock group, the expression levels of IL-10 in three groups pretreatment with CORM-2 at the doses of 2, 4 and 6 mg/kg were significantly increased (all P<0.05), and the expression levels of TGF-β were increased in two groups pretreatment with CORM-2 at the doses of 4 and 6 mg/kg (all P<0.05). The expression of IFN-γ in group pretreatment with CORM-2 at 6 mg/kg was significantly decreased compared with the shock group ( P<0.05). Conclusions:CORM-2 inhibited the activation of type 1 helper T cells to decrease the expression of proinflammatory factors and upregulated the expression of anti-inflammatory factors. Thus, CORM-2 reduced gut inflammation and protected the intestinal barrier.

Objective To investigate the influencing factors for the prognosis of adult patients with chronic drug-induced liver injury (DILI). Methods A total of 255 patients who were diagnosed with chronic DILI by liver biopsy in The Fifth Medical Center of Chinese PLA General Hospital from January 2014 to December 2018 were enrolled, and according to the liver function after 2 years, they were divided into non-recovery group and recovery group. The two groups were analyzed in terms of the clinical data including age, sex, body mass index, types of drugs used, type of DILI injury, severity of DILI injury, underlying diseases, laboratory markers, liver histology, and 2-year prognosis. The t -test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test was used for comparison of categorical data between two groups. Univariate and multivariate logistic regression analyses were used to investigate the independent risk factors for the prognosis of chronic DILI. Results After 2 years of follow-up, 195 patients (76.5%) achieved the recovery of liver function, while 60 patients (23.5%) did not achieve such recovery. There were significant differences between the two groups in the type of DILI injury ( P =0.028), the proportion of patients with diabetes ( P =0.048), and the degree of liver fibrosis ( P < 0.001), and compared with the recovery group, the non-recovery group had significantly higher levels of baseline white blood cell count, platelet count (PLT), alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transpeptidase, and total bile acid and a significantly lower level of cholinesterase (ChE) (all P < 0.05). The baseline characteristics were included in the univariate logistic regression analysis, and the results showed that PLT, ALT, AST, ChE, and fibrosis degree were significantly associated with the prognosis of chronic DILI (all P < 0.05). The multivariate logistic regression analysis of the above variables showed that PLT < 100×10 9 /L (odds ratio [ OR ]=3.592, 95% confidence interval [ CI ]: 1.128-11.438, P =0.003) and ALT > 2×upper limit of normal (ULN) ( OR =3.080, 95% CI : 1.331-7.127, P =0.009) were independent risk factors for the prognosis of chronic DILI. Conclusion When patients meet the diagnostic criteria for chronic DILI, the independent risk factors PLT < 100×10 9 /L and ALT > 2×ULN may be used to screen out the patients who are more likely to have poor prognosis.