AIM:The effect of salvianolic acid B on myocardial ischemia-reperfusion injury(MIRI)and its possible mechanism of action.METHODS:In this experiment,human induced pluripotent stem cell-derived cardiomyo-cytes(hiPSC-CMs)were used to establish a myocardial injury model by hypoxia for 6 hours and reoxygenation for 24 hours.Salvianolic acid B(7.5,15,30,60,120 μg/mL)was selected Study the effects of 5 dose groups on the imped-ance and field potential of hiPSC-CMs and hiPSC-CMs myocardial injury models.Subsequently,animal experiments were conducted and divided into sham surgery group,myocardial ischemia-reperfusion model group,salvianolic acid B low(15 mg/kg),medium(30 mg/kg),high(60 mg/kg)dose group,and positive control propranolol(2.5 mg/kg)group.After 4 days of gastric administration,left anterior descending coronary artery ligation surgery(ischemia for 30 minutes,reperfu-sion for 24 hours)was performed to establish a rat MIRI model,and the ischemic area of the rat heart was observed using TTC staining method.HE staining method was used to observe the pathological morphology of rat myocardial structure.Ex-ploring the protective effect of salvianolic acid B on MIRI by measuring serum central muscle enzyme activity using a fully automated biochemical analyzer.Use a reagent kit to detect the possible antioxidant mechanisms involved in the protection of MIRI by salvianolic acid B.RESULTS:Salvianolic acid B has the effect of enhancing the contractility of hiPSC CMs,reducing myocardial injury,and improving field potential function.And it can effectively reduce the myocardial ischemic area of the rat MIRI model,improve myocardial structural damage,reduce myocardial enzyme activity,and protect the heart.In addition,it can also reduce the content of malondialdehyde(MDA)in the serum of MIRI rats,enhance the activ-ity of superoxide dismutase(SOD)and reduced glutathione(GSH),and play an antioxidant role in improving myocardial injury.CONCLUSION:Salvianolic acid B can effectively reduce MIRI and play a role in improving the physiological function of myocardial cells and protecting the heart.Its mechanism of action may be related to reducing MDA levels,in-creasing SOD activity,and enhancing GSH activity of the serum.

Objective:To investigate the role of coupler perfusion in transrectal ultrasound in the diagnosis of preoperative T staging of rectal cancer.Methods:A retrospective analysis of the preoperative clinical data of 132 patients with rectal cancer in the People′s Hospital of Inner Mongolia Autonomous Region from June 2015 to November 2020. According to whether or not the patients agreed to coupler perfusion before ultrasound examination, they were divided into 2 groups, namely the perfusion group 69 cases and the non-perfusion group of 63 cases, with postoperative pathology as the gold standard, and compared with magnetic resonance imaging(MRI) to evaluate the accuracy of the 2 groups and MRI in the T staging of rectal cancer.Results:The total coincidence rates of the coupling agent perfusion group, non-perfusion group and MRI group for the diagnosis of rectal cancer T staging were 89.9%, 76.2% and 87.9%, respectively, and the difference among the three methods was statistically significant (χ 2=6.096, P=0.047). The diagnostic sensitivity of the coupling agent perfusion group for T1 stage was 96.0%, which was higher than 61.5% of the non-perfusion group and 92.3% of the MRI ( P=0.010). The specificity of the perfusion group for the diagnosis of T2 stage was 95.7%, higher than the non-perfusion group and MRI ( P=0.037), the positive predictive value of the perfusion group for T2 stage was 90.9%, which was higher than the non-perfusion group and MRI ( P=0.035). The diagnostic accuracy of the perfusion group for T2 stage was 94.2%, higher than the non-perfusion group and MRI (χ 2=7.070, P=0.029). There were no statistically significant differences in diagnostic sensitivity, specificity, positive predictive value, negative predictive value and accuracy among the perfusion group and the non-perfusion group and the MRI for T3 and T4 (all P>0.05). Conclusions:Coupled-agent perfusion makes it convenient and fast for intracavity ultrasound to diagnose T staging of rectal cancer, and the diagnostic efficiency is comparable to MRI. In particular, it can be used as a highly reliable imaging method for T1 and T2 rectal cancer.

Objective:To assess the value of amide proton transfer weighted (APTw) imaging in the evaluation of pH changes in infarct core (IC) and ischemic penumbra (IP) in subacute cerebral infarction.Methods:The data of twenty-three subacute cerebral infarction patients with unilateral steno-occlusive disease of the middle cerebral artery (subacute infarction group) from April to November 2019 in the First Affiliated Hospital of Dalian Medical University were prospectively analyzed. Fifteen healthy volunteers were enrolled in this study as the control group. All subjects underwent conventional MRI, DWI, 3D-pseudo continuous arterial spin labeling (3D-pCASL) and APTw sequences. Based on DWI images, relative cerebral blood flow (rCBF) and APTw images to determine the region of IC, blood flow penumbra [cerebral blood flow(CBF)-DWI mismatch area, IP CBF] and metabolic penumbra (APTw-DWI mismatched area, IP APT). 3D ROIs were used to semi-automatically measure the APTw signals and the volume of IC and IP CBF of the patients in subacute infarction group. The comparison of APTw signals between the infarct side and the contralateral side in the subacute infarction group, the comparison of bilateral APTw signals in the control group, and the comparison of APTw signals in the IC and IP CBF regions were performed by paired-sample t test or Wilcoxon signed-rank test. The paired-sample t test or Mann-Whitney U test was used to compare the APTw signals between the two groups. The Friedman test was applied to compare the difference of volumes among IP CBF1.5, IP CBF2.5 and IP APT . Results:There was no significant difference of the APTw signals among the IC, the contralateral side in the subacute infarction group and the control group ( P>0.05). The APTw signals of IP CBF and IC of the infarction group were statistically different ( P<0.05). Compared with the contralateral side of IP CBF1.5 (3.7±1.7, -1.84±1.48, 5.57±2.75), the APTwmax (3.07±1.41, t=-3.012, P=0.006), APTw min [-1.30 (-1.74, -0.57), Z=-2.099, P=0.036], and APTwmax-min(4.51±2.58, t=-3.273, P=0.003) signals in the IP CBF1.5 were decreased ( P<0.05). Compared with the contralateral side of IP CBF2.5 [-1.53 (-2.80, -0.91), 5.31±2.61], the APTw min [-1.08 (-1.60, -0.49), Z=-2.616, P=0.009] and APTwmax-min (4.41±2.72, t=-3.228, P=0.004) signals in the IP CBF2.5 were decreased. The volumes of IP CBF1.5 [107.51(50.08, 138.61)mm 3], IP APT [99.00 (53.27, 121.335) mm 3] and IP CBF2.5 [89.91 (51.53, 139.87) mm 3] were successively reduced (χ2=7.913, P=0.019), and the volume of IP CBF2.5 was significantly smaller than that of IP CBF1.5 ( P=0.037). Conclusion:The acid-base metabolism in the IC of subacute cerebral infarction is not obvious, but the blood flow penumbra has local acid-base metabolism imbalance, and the range of metabolic penumbra coincides with the blood flow penumbra.

Objective:To investigate the regulatory effects and mechanism of mannose-binding lectin(MBL) on autophagy during the differentiation of 3T3-L1 adipocytes, and provide the feasibility for targeting autophagy to prevent obesity and related pathological conditions in natural immunity.Methods:3T3-L1 preadipocytes were cultured in vitro and induced to differentiation. Cell differentiation and lipid accumulation were analyzed by oil red O staining and CCK-8 was used to detect the effect of different concentrations of MBL (0, 1, 5, 10 μg/ml) on cell proliferation ability at different differentiation stages. Western blot was used to analyze the expression of MBL(10 μg/ml) on the key autophagy factors LC3B, Beclin1 and p62 protein at different stages of differentiation, and the changes of lipid droplet accumulation under the intervention of MBL were observed by oil red O staining. The protein and mRNA expression of autophagy key factors under the intervention of different concentrations of MBL were detected by Western blot and qRT-PCR. And autophagy flow analysis based on autophagic degradation was used to further illustrate the autophagic activity. The expression and phosphorylation of adenosine monophosphate-activated protein kinase(AMPK)/mammalian target of rapamycin(mTOR) signaling molecules were analyzed by Western blot. Results:The results of oil red O staining showed that 3T3-L1 preadipocytes could achieve complete differentiation after 10 days of induction. CCK-8 showed that the concentration of MBL (1-10 μg/ml) in the experimental group had no effect on cell proliferation at different differentiation stages. During the differentiation of 3T3-L1 preadipocytes, Western blot and qRT-PCR showed that the expression of autophagy-related proteins and mRNA levels was enhanced in the MBL treated group, and presented a concentration-dependent relationship. Oil red O staining showed that the lipid droplets in adipocytes at different stages of differentiation are reduced to varying degrees under the intervention of MBL. Fluorescence microscopy results further confirmed that MBL enhanced the autophagy activity of adipocytes by increasing the synthesis of autophagosomes. Moreover, under the intervention of MBL, the phosphorylation level of AMPK was significantly up-regulated, while the phosphorylation level of mTOR was significantly down-regulated, also showing a concentration-dependent relationship.Conclusions:MBL accelerates the autophagy process during the differentiation of 3T3-L1 adipocytes through AMPK/mTOR signaling pathway, reduces lipid accumulation, providing a possible functional pathway for the treatment of obesity and related metabolic diseases.

Objective:To investigate the regulatory effects and mechanism of mannan-binding lectin (MBL) on adipogenic differentiation of 3T3-L1 preadipocytes.Methods:3T3-L1 preadipocytes were induced to differentiate into adipocytes in vitro, and stimulated with different concentrations of MBL (0, 1, 10, 20 μg/ml). Firstly, changes in cell proliferation ability were detected by CCK-8. Then lipid accumulation was analyzed by Oil red O staining and intracellular triglyceride content determination. Further, the expression of adipogenic differentiation-related factors PPARγ and C/EBPα at protein and mRNA levels were detected by Western blot and qRT-PCR, respectively. Finally, Western blot was used to analyze the expression and phosphorylation of Akt, a signal molecule related to adipogenic differentiation. Results:MBL at the concentrations of 0, 1, 10 and 20 μg/ml had no effect on the proliferation of 3T3-L1 preadipocytes. The level of triglyceride in MBL treatment groups decreased in a dose-dependent manner on 3 d after 3T3-L1 preadipocyte differentiation. Results of Oil red O staining showed that the number of lipid droplets in MBL treatment groups reduced significantly, and the absorbance values also decreased significantly in a concentration-dependent manner. Western blot and qRT-PCR results showed that the expression of PPARγ and C/EBPα at both protein and mRNA levels in MBL treatment groups decreased significantly in a dose-dependent manner, and the phosphorylation level of Akt was significantly down-regulated as well.Conclusions:MBL regulates the adipogenic differentiation of 3T3-L1 preadipocytes via Akt signaling pathway.

Objective: To summarize the clinical characteristics of a patient with cerebellar ataxia, areflexia, pes cavus, optic atrophy and sensorineural hearing loss (CAPOS) syndrome, followed by relative literature review. Methods: The medical history, physical examination and results of relative auxiliary examinations were collected from a CAPOS syndrome patient, who was definitely diagnosed by gene detection. Results: The patient was a 20-year-old male, complaining of poor coordination for 19 years, impaired vision for 15 years and hearing loss for 13 years. When he was eleven months old, weakness of four limbs happened after diarrhea but recovered spontaneously a few days later. Then his poor coordination was discovered. His vision has decreased progressively since the age of five and he began to suffer from bilateral hearing loss after fever at the age of seven. Anti-infectious and immunoregulatory treatment was ineffective at that time. Physical examination showed that bilateral visual acuity decreased. Transient horizontal gaze-evoked nystagmus and bilateral hearing loss were detected. Obvious shaking was observed with closed eyes and toes together. Finger-to-nose, finger tracking, heel-knee-tibia and alternate motion tests were slightly inaccurate. Deep tendon reflexes disappeared and no pes cavus was observed. Pure tone audiometry revealed bilateral sensorineural hearing loss. Cranial magnetic resonance imaging indicated bilateral optic atrophy. ATP1A3 gene detection in the patient showed c. 2452G>A (p. Glu818Lys) heterozygous mutation while his parents were detected no such mutation in the same locus. Conclusions As for young patients who suffer from acute cerebellar ataxia after fever, disappeared tendon reflexes, atrophy of optic nerves or sensorineural hearing loss, they should be alerted to CAPOS syndrome when immunomodulating or anti-inflammatory therapy has been proved to be useless. Positive family history and ATP1A3 gene mutation would be beneficial to definite diagnosis.

From September to October 2018, a questionnaire survey was conducted on the status of diabetes management, basic equipment allocation, complication screening and follow-up, basic drug supply and health education among 136 grassroots institutions in Nanjing. Doctors in the 136 grassroots institutions who had received diabetes specialist training accounted for 18.2% (358/1 968) of total general practitioners. The management rate of diabetes patients was 98.67% (196 352/199 000) , and the standard management rate was 88.63%(174 024/196 352).The screening and follow-up rates of diabetic complications were 89.0%(121/136) and 61.2%(74/121) for glycosylated hemoglobin; 64.7%(88/136), 58.0%(51/88) for urine microalbumin;51.5% (70/136), 62.9%(44/70) for diabetic foot; 44.9%(61/136), 42.6%(26/61) carotid ultrasonography; 32.4%(44/136) and 59.1%(26/44) for fundus examination; 17.6%(24/136) and 33.3%(8/24) for ankle brachial index. The rates of screening for glycosylated hemoglobin, urinary microalbumin, diabetic foot, carotid B ultrasound, fundus and ankle brachial index in urban areas were significantly higher than those in the suburbs (both P<0.05). The diabetes health education was regularly conducted in 92.0%(126/136) of grassroots institutions. At present, the grassroots institutions in Nanjing have established health archives for most diabetic patients and conducted health education regularly, but the professional training and the screening of diabetic complications need to be strengthened.

Objective: To investigate the spatial relationship between intraparotid facial nerve and parotid ducts in different types of parotid neoplasms with 3D improved motion-sensitized driven-equilibrium pulse (3D-iMSDE) MR neurography, and to guide the operation plan. Methods: By means of prospective research methods, the sequences of 3D-iMSDE and 3D-T₂-fast field echo (3D-T2-FFE) were obtained in 30 healthy volunteers and 40 patients with parotid gland tumors, respectively. All patients with parotid mass were found by physical examination, by maxillofacial surgeons or by ultrasonography or other imaging examinations. Facial nerve trunk with its first branches and parotid ducts were evaluated independently on images of iMSDE and T₂-FFE sequence for volunteers. The signal intensity ratio (SIR) of both facial nerve and parotid ducts were compared respectively in two sequences The relationship between intraparotid facial nerve and parotid ducts with the lesion were analyzed on images of iMSDE and T₂-FFE sequences for 40 patients. The results were compared with intra-operative anatomy and post-operative pathological findings.Wilcoxon symbolic rank test, t test and chi-square test were used Results: The image scores of intra-parotid facial nerve and parotid ducts in iMSDE sequence were higher than that of T₂-FFE sequence in all 30 healthy volunteers. The difference was statistically significant (Z=-6.197,P<0.05). Both the facial nerve and parotid duct on images of iMSDE sequence had higher SIR than that of T2-FFE. The differences were statistically significant (t=10.772,11.586, respectively;P<0.05). Forty patients with parotid gland tumors had a total of 41 lesions. Compared with the intraoperative anatomy, the accuracy of iMSDE and T₂-FFE sequences in showing the relationship between the facial nerve and its primary branches with the mass were 36/41 (87.8%), and 22/41 (53.7%) respectively, and the accuracy of the two sequences in showing the relationship between the dominant duct of the parotid gland and the mass were 38/41 (92.7%) and 30/41 (73.2%), respectively.The accuracy rate in showing the relationship between facial nerve and its primary branch and the mass on images of iMSDE sequence were higher than that of T₂-FFE sequence, and also in showing the relationship of parotid duct and the mass. The statistical differences were significant (χ²=9.725, 5.513;P<0.05,respectively). In the iMSDE sequence, the relationship between the facial nerve and its first branches (temporofacial and cervicofacial division) with the tumor was correctly demonstrated in 36 cases. 38 cases of parotid ducts were satisfactorily displayed on image of iMSDE sequence. Conclusions The 3D-iMSDE MRN sequence can correctly show the relationship between intraparotid facial nerve and parotid duct and the parotid gland neoplasms.

Objective To construct a human renal epithelial cell line HEK293T by CRISPR-Cas9-based site-directed knock-in of vascular endothelial growth factor 165 (VEGF165) gene, and avoid the off-target effect caused by lentivirus infection. Methods The VEGF165 expression vector with homologous arm (pUCm-T-VEGF165 plasmid) and the sgRNA expression vector [pSpCas9(BB)-2A-Puro-sgRNA plasmid] were designed and constructed based on the DNA sequence of the EZH2 gene, and then co-transfected into HEK293T cells. The expression of VEGF165 mRNA was detected by qPCR and the expressions of VEGF165 proteins were detected by Western Blot. Results The qPCR and Western Blot results showed that, comparing with the control, the pUCm-T-VEGF165 plasmid and pSpCas9(BB)-2A-Puro-sgRNA plasmid, the expression of the co-transfection plasmid were significantly increased, i.e. 3.42±0.30 vs. 1.02±0.21, 1.13±0.16 and 0.98±0.18 for the VEGF165 mRNA level (all P<0.01), and 1.13±0.16 vs. 1.02±0.06, 0.88±0.03 and 0.80±0.05 for the VEGF165 protein level (all P<0.01), respectively. Besides, the expression of EZH2 was significantly down-regulated, i.e. 0.14±0.06 vs. 1.08±0.11, 1.02±0.12 and 1.13±0.16 for the EZH2 mRNA level (all P<0.01), and 0.23±0.03 vs. 1.05±0.13, 0.91±0.04 and 0.81±0.06 for the EZH2 protein level (all P<0.01), respectively. This result showed that the VEGF165 was successfully inserted into the EZH2 genome, interfering the EZH2 expression. Conclusions VEGF165 gene can be successfully knocked into HEK293T cells by CRISPR/Cas9 system.

Objective To evaluate the clinical efficacy and adverse reactions of S-1 chemotherapy plus high-low oxygen radiotherapy synchronously in treatment of patients with locally advanced pancreatic cancer. Methods Sixty-four patients with locally advanced pancreatic cancer in Rizhao People's Hospital and the Affiliated Hospital of Qingdao University from January 2013 to October 2015 were randomly divided into study group and control group by using envelope method, in which the study group was treated with oral administration of S-1 plus high-low oxygen radiation synchronously and the control group with intravenous gemcitabine chemotherapy. The efficiency, disease control rate, clinical benefit rate, distant metastasis rate, survival rate and adverse reactions of the two groups were compared. Results The effective rates of the study group and the control group were 70.4 %(19/27)and 32.1 %(9/28),the difference was statistically significant (χ2=8.04,P<0.01), and the disease control rates were 88.9 % (24/27) and 67.9 % (19/28) (χ2= 3.56, P >0.05). The clinical benefit rates of the study group and the control group were 77.8 % (21/27) and 57.1 % (16/28) (χ2=2.66,P >0.05), and the distant metastasis rates were 63.0 %(17/27) and 71.4 %(20/28) (χ2=0.45, P > 0.05). The 1-year survival rates of the study group and the control group were 63.0 % (17/27) and 32.1 % (9/28), and the 2-year survival rates were 37.0 % (10/27) and 10.7 % (3/28), the differences were statistically significant (χ2= 5.24, P < 0.05; χ2= 5.28, P< 0.05). While there were no significant difference in the incidence of serious adverse reactions between the two groups (P> 0.05). Conclusion The treatment of locally advanced pancreatic cancer with S-1 plus high-low oxygen radiotherapy synchronously is better than that with intravenous gemcitabine chemotherapy in terms of effective rate, 1-year survival rate and 2-year survival rate,with no increase of adverse reactions.