Objective:To evaluate the immunogenicity to prototype strain of 2019-nCoV and cross immune response to its variants in children aged 3-11 years after completing the primary immunization of a 2019-nCoV inactivated vaccine for 28-42 days.Methods:From January to July 2022, children aged 3-11 years who had been vaccinated with 2019-nCoV inactivated vaccine according to the (0, 28) day immunization schedule were recruited in Rushan, Shandong Province. On 28-42 days after the primary immunization, venous blood samples (3 ml) were collected from them for the detections of the levels of neutralizing antibody to prototype strain, Beta variant, Gamma variant and Delta variant and the neutralizing antibody positive rates and GMTs were calculated.Results:A total of 395 children were included in the immunogenicity analysis, including 212 children aged 3-5 years and 183 children aged 6-11 years. The positive rates of neutralizing antibody to prototype strain, Beta, Gamma, and Delta variants on 28-42 days after the primary immunization were 100.00%, 74.68%, 99.24%, and 97.22%, respectively, there were no significant differences between two age groups ( P>0.05). On 28-42 days after the primary immunization, the GMTs of neutralizing antibody to prototype strain, Beta , Gamma and Delta variants were 168.19, 10.51, 53.65, and 31.10, respectively, the differences between two age groups were not significant ( P>0.05). Conclusion:The immunization with 2 doses of 2019-nCoV inactivated vaccine in 3-11 years old children showed good immunogenicity and produced certain cross protection to 2019-nCoV variants.

Objective:To study the roles of hepatoma-derived growth factor (HDGF), vascular endothelial growth factor (VEGF) and mucin 5B (MUC5B) in hepatolithiasis associated with intrahepatic cholangiocarcinoma (HICC).Methods:The clinical data and tissue specimens of 116 patients who underwent hepatectomy at the Third Affiliated Hospital of Wenzhou Medical University from October 1999 to October 2019 were retrospectively analyzed. There were 41 patients with HICC (the HICC group), 38 patients with intrahepatic cholangiolithiasis (the intrahepatic cholangiolithiasis group), and 37 patients with benign liver tumor who underwent hepatectomy (the control group). There were 47 males and 69 females, with age of (66.1±3.2) years old. The positive expressions of HDGF, VEGF and MUC5B in the three groups were compared. In 41 patients with HICC, the correlation between positive expressions of HDGF, VEGF and MUC5B with patients’ clinical characteristics were studied.Results:Compared with the control group, the positive expression rates of HDGF, VEGF and MUC5B in the HICC group and the intrahepatic cholangiolithiasis group were significantly increased, ( P<0.05). Compared with the intrahepatic cholangiolithiasis group, the positive expression rates of HDGF, VEGF and MUC5B in the HICC group were significantly increased ( P<0.05). Positive expression of VEGF in HICC patients was correlated with tumor differentiation, tumor local invasion, tumor length, tumor stage, tumor carbohydrate antigen CA19-9 level and lymph node metastasis ( P<0.05). Spearman correlation analysis showed that HDGF was positively correlated with VEGF expression in HICC tissues specimens ( r=0.935, P<0.01). Survival analysis showed that the cumulative survival rates (36.7%, 17.1%, 7.3%) of patients with positive expression of VEGF were significantly lower than that of patients with negative expression (51.2%, 26.8%, 19.5%) at 1, 3, 5 years after surgery ( P<0.01). The cumulative survival rate (34.1%, 17.1%, 4.9%) of patients with MUC5B positive expression were significantly lower than that of patients with negative expression (53.7%, 31.7%, 17.1%) at 1, 3, 5 years after surgery ( P<0.01). Conclusion:HDGF can be used as a reference indicator for early assessment of HICC. Overexpressions of VEGF and MUC5B can be used as important indicators for HICC in evaluating disease progression and prognosis.

Objective:To explore the expression and clinical significance of mucin 5B in patients with primary intrahepatic bile duct stones (PHL) after hepatectomy.Methods:Collected the bile duct mucosa of 48 patients with intrahepatic bile duct stones (PHL group) and 16 patients with non-calculous benign liver disease (control group) who underwent hepatectomy in the Third Affiliated Hospital of Wenzhou Medical University from January 2014 to January 2019, Bile duct wall, bile and venous blood. The preoperative bile and serum indexes of the two groups were compared. Immunohistochemical staining was used to examine the expression of mucin 1 and mucin 5B in the bile duct wall, and the bile duct wall was examined pathologically by HE routine staining. With mucin 1 as a positive control and β-actin as an internal reference gene, real-time PCR was used to examine the mRNA expression levels of mucin 1 and mucin 5B in the bile duct mucosa. Pearson correlation analysis was used to analyze the correlation of variables within the PHL group.Results:The preoperative serum lipid indexes in the PHL group were higher than those in the control group, while the total bile acid concentration [(181.5±18.2) mmol/L vs. (192.1±22.5) mmol/L] and the molar percentage of bile acid [(80.7±1.6)% vs. (89.7±1.0)%] is lower than the control group, the difference is statistically significant ( P<0.05). The expression of mucin 1 mRNA in the PHL group was higher than that in the control group, but the difference was not statistically significant ( P>0.05). The expression of mucin 5B mRNA in the PHL group was significantly higher than that in the control group [(0.94±0.12) vs. (0.73±0.24)], the difference was statistically significant ( P<0.05); The increased expression of bile duct mucin 5B mRNA was negatively correlated with the level of total bile acids in bile ( r=-0.4, P<0.05). Conclusions:The increased expression of mucin 5B is closely related to PHL, which may be related to the promotion of bile acid absorption by the bile duct mucosal epithelium, which causes mucin to secrete into the bile in large quantities, leading to the formation of stone-causing bile.

Objective To investigate the relationship between Pst-Ⅰ polymorphism of mucin 4 gene and primary hepatolithiasis in the Chinese population.Methods Venous blood of 96 healthy controls and 56 patients with hepatolithiasis were collected,and DNA was extracted.Polymerase chain reaction combined with restriction enzyme (PCR-RFLP) digestion was used to detect Pst-Ⅰ polymorphism of mucin 4 gene in the two groups.The genotype and gene frequency between the two groups were then compared.Results The genotype frequencies of GC,GT,TT in the control and the hepatolithiasis groups were 21.3％,12.7％,55.6％ and 53.2％,41.2％,19.8％,respectively.The alleles C and T gene frequencies in the control and the hepatolithiasis groups were 21.5％,72.7％ and 66.3％,30.2％,respectively.There were significant differences in Pst-Ⅰ polymorphism of mucin 4 genotype frequency and gene frequency between the two groups.Conclusion The data showed Pst-Ⅰ polymorphism of mucin 4 gene was associated with primary hepatolithiasis in Chinese patients.

Objective To investigate the function of mucin 2,mucin 4 on formation of lithogenic bile in patients with calculus of intrahepatic duct.Methods Bile duct mucosa,bile duct wall,bile and plasma were collected from 56 patients with calculus of intrahepatic duct (CID group) and 17 individuals without calculus of intrahepatic duct (control group).The bile duct wall was stained with mucin 2 (MUC2) and mucin 4 (MUC4).Reverse-transcription polymerase chain reaction (RT-PCR) was applied to study the mRNA expressions of MUC2 and MUC4 in the bile duct mucosa.The correlation of the bile duct and serum lipid index was analyzed.Results Serum lipid index in the CID group was significantly higher than control group (all P＜0.05).Biliary total bile acids and bile acids mol percentage were lower in the CID group than control group (both P＜0.05).The expressions of MUC2 was not increased significantly in CID group than the control group (all P＞0.05).The expressions of MUC4 were more significantly increased in CID group than the control group (P＜0.05).The mRNA of MUC4 in the CID group was also more significantly increased than in control group (P＜0.01).There were no correlations between MUC4 expression and the level of biliary total bile acid in the CID group (r=-0.374,P＜0.05).Conclusion The expression of MUC4 in patients with CID was enhanced,which promoted the absorption of bile acid by the mucosal epithelium of the bile duct,and caused a large amount of mucin to be secreted into bile,which may be related to the formation of stony bile.

Objective To evaluate the role of the angiotensin Ⅱ type 2 receptor (AT2R) in repeated propofol anesthesia-induced neuroapoptosis in the basal ganglia of newborn rats.Methods Fiftyfour pathogen-free Sprague-Dawley rats,aged 7 days,weighing 10-15 g,were divided into 3 groups (n=18 each) using a random number table:control group (group C),repeated propofol anesthesia group (group P) and AT2R agonist CGP42112A group (group G).In group C,0.9％ sodium chloride injection 3 ml/kg was intraperitoneally injected,and half of the initial dose 1.5 ml/kg was given every 20 min for 5 times in total,lasting for 3 consecutive days.In group P,propofol 30 mg/kg was intraperitoneally injected,and half of the initial dose 15 mg/kg was given every 20 min for 5 times in total,lasting for 3 consecutive days.In group G,CGP42112A 1 mg/kg was intraperitoneally injected,propofol 30 mg/kg was intraperitoneally injected 5 min later,and half of the initial dose of propofol 15 mg/kg was given every 20 min for 5 times in total,lasting for 3 consecutive days.Six rats were sacrificed at 2 h after emergence from anesthesia,and brains were removed for detection of neuroapoptosis in the basal ganglia by TUNEL assay.The apoptosis index was calculated.Another 6 rats were sacrificed,and the basal ganglia were isolated from brains to detect the expression of activated caspase-3,AT2R and peroxisome proliferator-activated receptor gamma (PPARγ) (by Western blot) and the expression of AT2R and PPARγ mRNA (by real-time polymerase chain reaction).The other 6 rats were fed until 28 days old,and the cognitive function was then assessed using Morris water maze test.Results Compared with group C,the escape latency was significantly prolonged,the time of staying at the target quadrant was shortened,the frequency of crossing the platform was decreased,the apoptosis index of the basal ganglia was increased,the expression of activated caspase-3 was up-regulated,and the expression of AT2R and PPARγprotein and mRNA was down-regulated in group P (P＜0.05),and no significant change was found in the parameters mentioned above in group G (P＞0.05).Compared with group P,the escape latency was significantly shortened,the time of staying at the target quadrant was prolonged,the frequency of crossing the platform was increased,the apoptosis index of the basal ganglia was decreased,the expression of activated caspase-3 was down-regulated,and the expression of AT2R and PPARγ protein and mRNA was up-regulated in group G (P＜0.05).Conclusion Inhibited activation of AT2R is involved in repeated propofol anesthesia-induced neuroapoptosis in the basal ganglia of newborn rats.

Objective To evaluate the effect of hydromorphone postconditioning on the electrophysiological stability during ischemia-reperfusion (I/R) in isolated rat hearts.Methods Healthy adult male Sprague-Dawley rats,aged 2-3 months,weighing 280-360 g,were heparinized and anesthetized with pentobarbital sodium.Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95％ O2-5％ CO2 at 37 ℃.Twenty-four Langendorff-perfused hearts were divided into 3 groups (n =8 each) using a random number table:control group (group C),I/R group and hydromorphone postconditioning group (group HM).The isolated hearts were subjected to 60 min ischemia followed by 60 min reperfusion to establish the model of isolated heart I/R injury.The isolated hearts were perfused with K-H solution containing 4.1 ng/ml hydromorphone for 10 min starting from onset of reperfusion in group HM.Heart rate,electrocardiogram,coronary flow,and monophasic action potential amplitude,in the left ventricular endocardium,mid-myocardium and epicardium the maximal increase rate (Vmax) at the 0 phase,and monophasic action potential duration at 50％ and 90％ repolarization (MAPD50 and MAPD90,respectively) were recorded at 20 min of stabilization (T0) and 10,25,40 and 60 min of reperfusion (T1-4).The transmural dispersion of repolarization (TDR) was calculated,and the time for restoratiou of spontaneous heart beat was recorded.Results There was no significant difference in the heart rate,coronary flow or monophasic action potential amplitude between the three groups (P＞0.05).Compared with group C,V in the epieardium was significantly decreased,MAPD50 and MAPD90 in the three transmural layers were prolonged,and TDR was prolonged in group I/R (P＜0.05).Compared with group I/R,the time for restoration of spontaneous heart beat,MAPD50 in the endoeardium and mid-myocardium,and MAPD90 and TDR in the endocardium were significantly shortened (P＜0.05),and no significant change was found in V in group HM (P＞0.05).Conclusion Hydromorphone postconditioning is helpful in maintaining the electrophysiological stability during I/R in isolated rat hearts.

Objective To evaluate the role of δ-opioid receptors in hydromorphone postcondition-ing-induced maintenance of electrophysiological stability during ischemia-reperfusion(I∕R)in isolated rat hearts. Methods Healthy male Sprague-Dawley rats, aged 2-3 months, weighing 280-360 g, were used in this study. The animals were anesthetized with intraperitoneal pentobarbital 60 mg∕kg. Their hearts were immediately removed and perfused in a Langendorff apparatus. Thirty-two isolated hearts were divided into 4 groups after successful preparation of Langendorff perfusion model(n=8 each)using a random number ta-ble: control group(group C), group I∕R, hydromorphone postconditioning group(group HP)and hydro-morphone plus δ-opioid receptor antagonist naltridole postconditioning group(group HNP). In HP and HNP groups, the hearts were perfused for 10 min with K-H solution containing 41 ng∕ml hydromorphone and 41 ng∕ml hydromorphone plus 5 μmol∕L naltridole, respectively, and then with K-H solution for 50 min. At 20 min of stabilization(T0)and 10, 25 and 60 min of reperfusion(T1-2), heart rate(HR), monophasic action potential(MAP)duration at 90% repolarization(MAPD90)of the two layers(endocar-dium, epicardium)of the anterior left ventricular wall were recorded. Transmural dispersion of repolariza-tion(TDR)was calculated. The development of arrhythmia, time for restoration of spontaneous heart beat and duration of arrhythmia were recorded during the period of reperfusion. Results Compared with group C, MAPD90of endocardium at T1-2and MAPD90of epicardium at T1were significantly prolonged in I∕R and HP groups, HR was significantly decreased at T2-3, MAPD90of endocardium and epicardium was prolonged at T1-3in group HNP, TDR was significantly enlarged at T1in group I∕R and at T2in group HNP, and TDR was decreased at T3in group HP(P<005). Compared with group I∕R, no significant change was found in arrhythmia score(P>005), the time for restoration of spontaneous heart beat was significantly shortened, and TDR was decreased at T1in HP and HNP groups, duration of arrhythmia was significantly shortened, and MAPD90of endocardium was shortened at T1in group HP, and HR was significantly decreased at T2-3, MAPD90of endocardium and epicardium was prolonged at T1-3, and TDR was decreased at T2-3in group HNP(P<005). Compared with group HP, no significant change was found in time for restoration of spon-taneous heart beat, duration of arrhythmia or arrhythmia score(P>005), HR was significantly decreased at T2-3, MAPD90of endocardium and epicardium was prolonged at T1-3, and TDR was increased at T3in group HNP(P<005). Conclusion The mechanism underlying hydromorphone postconditioning-induced maintenance of electrophysiological stability during I∕R is related to activating δ-opioid receptors in isolated rat hearts.

Objective To evaluate the role of the angiotensin Ⅱ type 2 receptor (AT2R) in repeated propofol anesthesia-induced neuroapoptosis in the hippocampus of newborn rats.Methods Fiftyfour pathogen-free Sprague-Dawley rats,aged 7 days,weighing 10-15 g,were divided into 3 groups (n=18 each) using a random number table:control group (group C),repeated propofol anesthesia group (group P) and AT2R agouist CGP42112A group (group G).In group C,0.9％ sodium chloride injection 3 ml/kg was intraperitoneally injected,and half of the initial dose 1.5 ml/kg was given every 20 min for 5 times in total,lasting for 3 consecutive days.In group P,propofol 30 mg/kg was intraperitoneally injected,and half of the initial dose 15 mg/kg was given every 20 min for 5 times in total,lasting for 3 consecutive days.In group G,a single bolus of CGP42112A 1 mg/kg was intraperitoneally injected,propofol 30 mg/kg was intraperitoneally injected 5 min later,and half of the initial dose of propofol 15 mg/kg was given every 20 min for 5 times in total,lasting for 3 consecutive days.At 2 h after emergence from anesthesia,6 rats were sacrificed and brains were removed for detection of neuroapoptosis in the hippocampus by TUNEL assay.The apoptosis index was calculated.Another 6 rats were sacrificed,brains were removed and hippocampi were isolated for determination of the expression of activated caspase-3,AT2R and peroxisome proliferator-activated receptor gamma (PPARγ) in hippocampal tissues by Western blot.The other 6 rats were fed until 28 days old,and the cognitive function was then assessed using Morris water maze test.Results Compared with group C,the escape latency was significantly prolonged,the time of staying at the target quadrant was shortened,the frequency of crossing the platform was decreased,the apoptosis index was increased,the expression of activated caspase-3 was up-regulated,and the expression of AT2R and PPARγ was down-regulated in group P (P＜0.05),and no significant change was found in the parameters mentioned above in group G (P＞0.05).Compared with group P,the escape latency was significantly shortened.the time of staying at the target quadrant was prolonged,the frequency of crossing the platform was increased,the apoptosis index was decreased,the expression of activated caspase-3 was down-regulated,and the expression of AT2R and PPARγ was up-regulated in group G (P＜0.05).Conclusion Inhibited activity of AT2R is involved in repeated propofol anesthesia-induced neuroapoptosis in the hippocampus of newborn rats.

OBJECTIVETo compare the effects between plum-blossom needle tapping at Jiaji (EX-B 2) combined with rehabilitation and simple rehabilitation for ataxia of cerebellar apoplexy.

METHODSSixty patients with ataxia of cerebellar apoplexy were randomly divided into an observation group,and a control group,30 cases in each one. Conventional rehabilitation training was applied in the control group. While based on the treatment as that of the control group, plum-blossom needle tapping at Jiaji (EX-B 2) was used in the observation group. All the treatment was given for 4 weeks. International cooperative ataxia rating scale (ICARS), Berg balance scale (BBS) and Barthel index (BD were observed before treatment,after 2-week treatment and 4-week treatment. Also, clinical effects After treating for 2 weeks and for 4 weeks, the results of ICARS,BBS and BI were obvi were compared.

RESULTSAfter treating for 2 weeksand for 4 weeks, the results of ICARS, BBS and BI were obviously better than those before treatment in the two groups (all P<0.05). After 2-week treatment,there was no sta tistical significance between the two groups (all P>0. 05). After 4-week treatment,all the scores of the observation group were apparently superior to those of the control group (all P<0. 05). The total effective rate was 96. 7% (29/30) in the observation group, better than 80. 0% (24/30) in the control group after 4-week treatment (P<0. 05). CCONCLUSION:Plum-blossom needle tapping at Jiaji(EX-B 2) combined with rehabilitation training markedly improve the coordination and balance functions and activities of daily life, which are better than simple rehabilitation.

Acupuncture Points ; Acupuncture Therapy ; methods ; Aged ; Ataxia ; etiology ; rehabilitation ; therapy ; Combined Modality Therapy ; Female ; Humans ; Male ; Middle Aged ; Needles ; Stroke ; complications ; Treatment Outcome