OBJECTIVE: To observe the effect of acupuncture on chondrocyte autophagy in rats of knee osteoarthritis (KOA) and explore its underlying mechanisms. METHODS: Forty male SPF-grade SD rats were randomized into a blank group, a model group, a suspension group, an acupuncture group, and a combined therapy group, 8 rats in each one. Except the blank group, KOA model was prepared by the injection with papain. The suspension exercise therapy (10 min each time, three times daily), acupuncture (at "Yanglingquan" [GB34], "Zusanli" [ST36], and "Dubi" [ST35] on the right side, 30 min each intervention, once daily) and the combined therapy (the suspension exercise therapy combined with acupuncture) were delivered in the suspension group, the acupuncture group and the combined therapy group, respectively. The intervention of each group was performed continuously for 6 days, and 4 consecutive weeks, at the interval of 1 day. Before and after intervention, Lequesne MG score was assessed in the rats. After intervention, HE staining was adopted to observe the cartilaginous tissue morphology of the right knee joints, and Mankin score was evaluated; the serum levels of interleukin (IL)-1β, IL-6, and tumor neurosis factor-α (TNF-α) were measured using ELISA; the real-time PCR was provided to determine the mRNA expression of collagen protein type Ⅱ(COL2), collagen protein type Ⅹ (COL10), phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt), mammalian target of rapamycin (mTOR), and autophagy-regulated protein (Beclin-1) in the cartilaginous tissue of the right knee joint; Western blot was employed to detect the protein expression of PI3K, phosphorylated PI3K (p-PI3K), Akt, phosphorylated Akt (p-Akt), mTOR, phosphorylated mTOR (p-mTOR) and Beclin-1 in the cartilaginous tissue of the right knee joint. RESULTS: Compared with the blank group, the rats in the model group showed the higher Lequesne MG score (P<0.01), thinner cartilage of the right knee, reduced chondrocytes and disordered arrangement, and higher Mankin score (P<0.01). Besides, in the model group, the serum levels of IL-1β, IL-6 and TNF-α were elevated (P<0.01), the mRNA expression of COL2 and Beclin-1 and the protein expression of Beclin-1 decreased (P<0.01), the mRNA expression of COL10, PI3K, Akt and mTOR, and the protein expression of p-PI3K, p-Akt and p-mTOR increased (P<0.01) in the cartilaginous tissue of the right knee joint. Compared with the model group, in the suspension group, the acupuncture group and the combined therapy group, the Lequesne MG scores were reduced (P<0.01), the cartilage of the right knee was thickened, the arrangement of chondrocytes was improved, and the Mankin scores were lower (P<0.01). Besides, in these intervention groups, the serum levels of IL-1β, IL-6 and TNF-α were reduced (P<0.01), the mRNA expression of COL2 and Beclin-1 and the protein expression of Beclin-1 increased (P<0.05, P<0.01), the mRNA expression of COL10, PI3K, Akt and mTOR, and the protein expression of p-PI3K, p-Akt and p-mTOR decreased (P<0.05, P<0.01) in the cartilaginous tissue of the right knee joint. When compared with the suspension group and the acupuncture group, in the combined therapy group, the Lequesne MG score was reduced (P<0.01), and the Mankin score was reduced (P<0.05, P<0.01). Besides, the serum levels of IL-1β, IL-6 and TNF-α were reduced (P<0.05), the mRNA expression of COL2 and Beclin-1 and the protein expression of Beclin-1 increased (P<0.05), the mRNA expression of COL10, PI3K, Akt and mTOR, and the protein expression of p-PI3K, p-Akt and p-mTOR decreased (P<0.05, P<0.01) in the cartilaginous tissue of the right knee joint. CONCLUSION Acupuncture can promote cartilage regeneration of knee joint and autophagy in KOA rats, alleviate inflammation, so as to retard cartilage degeneration, which may be possibly associated with the PI3K/Akt/mTOR signaling pathway.
Animals ; Male ; Acupuncture Therapy ; Proto-Oncogene Proteins c-akt/genetics* ; TOR Serine-Threonine Kinases/genetics* ; Osteoarthritis, Knee/physiopathology* ; Phosphatidylinositol 3-Kinases/genetics* ; Rats ; Signal Transduction ; Rats, Sprague-Dawley ; Chondrocytes/metabolism* ; Humans ; Autophagy ; Acupuncture Points

Objective:To investigate the effects of glycopyrrolate on intestinal spasm and hemodynamics in painless colonoscopy.Methods:A total of 100 patients who were scheduled to undergo painless colonoscopy were selected as the study subjects and randomly divided into two groups by a computerized number method. Ten patients in both groups dropped out because of disruption of the study protocol, and 45 patients from each group were included in the final analysis. Before anesthesia induction, patients in group glycopyrrolate (group G) were injected with 0.2 mg glycopyrrolate, while those in congtrol group (group C) were injected with an equal amount of saline. The heart rate, systolic blood pressure, and diastolic blood pressure were recorded at T 0 (baseline period), T 1 (after anesthesia induction), T 2 (colonoscopy over sigmoid colon), T 3 (colonoscopy over the liver region), T 4 (after the end of examination), and T 5 (at the awakening phase), and the degree of intestinal spasm was assessed intraoperatively using the Likert′s four-point scale. The numerical rating scale (NRS) was used to assess preoperative and postoperative pain. The incidence of adverse events was recorded. Results:The general data at baseline were not statistically different between the two groups ( P>0.05). During the procedure, patients in group G had lower intraoperative intestinal spasm scores than those in group C ( P=0.028). Intraoperative hypotension and bradycardia occurrence were lower in group G than in group C ( P<0.05), and intraoperative norepinephrine use was also lower than in the group C ( P=0.034). Postoperative visual analog scale pain scores were lower in group G ( P=0.047), but patients who used glycopyrrolate had a higher proportion of dry mouth ( P=0.035). Conclusion:During painless colonoscopy, preoperative administration of glycopyrrolate significantly improved intraoperative hemodynamic fluctuations, reduced the incidence of hypotension and bradycardia, and relieved postoperative pain. However, glycopyrrolate use resulted in the risk of dry mouth.

Objective:To study the value of sound touch elastography (STE) linear combined with ultrasound score (US) in the diagnosis of chronic hepatitis B (CHB) liver fibrosis, and to investigate whether their combination can improve the diagnostic efficiency of subdividing the degree of CHB liver fibrosis. Furthermore, a comparison with STE linear combined with the serological model was performed to seek the optimal linear combination model.Methods:A total of 313 subjects were enrolled from September 2018 to December 2021 in Shenzhen Third People′s Hospital Affiliated to Guangdong Medical University, including 259 patients with CHB who had completed liver biopsy and 54 healthy volunteers. CHB patients were divided into liver fibrosis group (F1-F4 group) according to METAVIR classification standard, and healthy volunteers were used as the control group. All subjects underwent liver ultrasound examination, STE and blood biochemical indexes of liver function. The US was performed according to the liver ultrasound examination, and the liver stiffness measurement (LSM) was measured by STE, aspartate aminotransferase and platelet ratio index (APRI) was calculated by blood biochemical index. Fisher discriminant analysis was used to establish the linear combination (LC) diagnostic marker of US and LSM, and the linear combination (LC2) diagnostic marker of LSM and APRI, successively. Spearman rank correlation coefficient was used to analyze the correlations between US, LSM, APRI, LC2, LC and pathological results. The ROC curves of US, LSM, APRI, LC2 and LC for diagnosing CHB liver fibrosis were plotted, and the diagnostic efficiency of above diagnostic markers was evaluated according to the accuracy, sensitivity, specificity and area under the ROC curve (AUC).Results:The formula for the linear combination of US and LSM was LC=0.986 0×US+ 0.166 7×LSM, and LC was highly positively correlated with pathological findings ( rs=0.851, P<0.001), higher than US, LSM, LC2 and APRI ( rs=0.825, 0.775, 0.802, 0.586, all P<0.001). LC showed the best diagnostic efficiency. The AUCs for diagnosing ≥F1, ≥F2, ≥F3 liver fibrosis and =F4 cirrhosis were 0.945, 0.911, 0.954, 0.955, respectively, which superior to the AUCs of US (0.913, 0.879, 0.934 and 0.916, respectively), the AUCs of LSM (0.860, 0.871, 0.934 and 0.952, respectively) and the AUCs of LC2(0.899, 0.883, 0.941, 0.946, respectively). Compared with US, the AUC of LC diagnosis of ≥F1, ≥F2, ≥F3 liver fibrosis and =F4 cirrhosis increased by 3.2%, 3.2%, 2.0% and 3.9%, respectively, with all significant differences ( P<0.05). Compared with LSM, the AUC of LC increased by 8.5%, 4.0%, 2.0% and 0.3%, respectively, with significant difference ( P<0.05) except for stage =F4 cirrhosis.Compared with LC2, the AUC of LC increased by 4.6%, 2.8%, 1.3% and 0.9%, respectively, and there were significant differences in the diagnosis of ≥F1 and ≥F2 liver fibrosis ( P<0.05). Moreover, the overall efficiency of LC2 was not significantly improved than LSM, the difference was not significant ( P>0.05). Conclusions:US, LSM, LC2 and LC can be used to diagnose the degree of CHB liver fibrosis, but LC is better than US or LSM and LC2 alone, especially in the subdivision of mild liver fibrosis, which is a promising new diagnostic marker to subdivide the degree of CHB liver fibrosis.

Objective: To assess the value of cyclin D1 immunocytochemistry combined with a small panel molecular analysis in indeterminate cytological diagnosis of Bethesda category Ⅲ-Ⅴ. Methods: A consecutive cohort of 96 thyroid FNA specimens with indeterminate diagnosis (TBSRTC category Ⅲ-Ⅴ) and available histopathologic follow-up data were collected between December 2018 and December 2021 in Department of Pathology, Beijing Hospital. The cases were evaluated by cyclin D1 immunocytochemistry and molecular testing of BRAF^V600E or a small panel of markers (BRAF, N-RAS, H-RAS, K-RAS and TERT) in the FNA specimens. The identification of the optimal cut-off point of cyclin D1 for the diagnosis of malignancy was evaluated using the receiver operating characteristic (ROC) curves and the assessment of the area under the ROC curve (AUC). The specificity, sensitivity, positive predictive value (PPV) and negative predictive value (NPV) of all these markers were evaluated with the crosstabs and significance was calculated. Results: Ninty-six patients with 96 thyroid nodules were enrolled, including 42 cases of TBSRTC-III, 10 cases of TBSRTC-IV and 44 cases of TBSRTC-V. There were 79 females and 17 males with a median age of 47 years (range, 25 to 75 years). A 7.5% cut-off value for positive cyclin D1 nuclear immunostaining in thyroid cells demonstrated 100% PPV, 57.1% NPV, 81.0% sensitivity and 100% specificity for thyroid malignancy diagnosis. The sensitivity of the BRAF^V600E mutation test or combined with a small panel test alone for thyroid malignancy diagnosis were 65.5% and 69.0% respectively. The sensitivity for thyroid malignancy diagnosis increased to 94.0% and 95.2% respectively when combining the cyclin D1 immunocytochemistry with the molecular test, and the specificities remained 100% and 91.7% respectively.The accuracy of cyclin D1 immunocytochemistry combined with a small panel of molecular test in detecting thyroid malignancy increased to 94.8% compared to using these markers alone. Conclusions: The addition of cyclin D1 immunocytochemistry and a small panel of molecular testing to FNA cytology can increase the sensitivity and NPV of cytology in indeterminate categories, and this supplementary approach provides a simple, accurate and convenient diagnostic method for reducing unnecessary thyroidectomies.
Adult ; Aged ; Humans ; Middle Aged ; Biopsy, Fine-Needle ; Cyclin D1/genetics* ; Molecular Diagnostic Techniques ; Thyroid Nodule/genetics* ; Male ; Female

【Objective】 To investigate the profile of Rh blood group antigen in pregnant women and hemolytic disease of the newborn (HDN) in Qingdao area. 【Methods】 10 597 pregnant women admitted in our hospital during October 2016 to February 2020 were selected and the ABO, Rh blood group system antigen (D, C, c, E, e) and the irregular antibody were detected, and positive antibody was further identified. The irregular antibody of Rh blood group in pregnant women was statistically analyzed according to the history of blood transfusion and pregnancy. Twelve HDN cases were studied, and the results of ABO, Rh blood group antigen and irregular antibody, antibody property identification, HDN test and blood routine test were retrospectively analyzed. 【Results】 Among 10 513 cases of Rh-positive pregnant women, the common phenotype was CCee>CcEe>Ccee>ccEE>ccEe; among 84 cases of Rh-negative pregnant wome, the common phenotype was ccee>Ccee> CCee> ccEE>ccEe. The positive rate of irregular antibody was 1.06% (112/10597) in 10 597 pregnant women, of which the Rh antibody was the highest, rated at 56.25% (63/112). For 64 pregnant women with positive antibodies, antibodies against Rh system were different from those against other systems when stratified by the history of blood transfusion (P<0.05) and pregnancy (P<0.05). Twelve neonates were diagnosed with Rh-HDN, with IgG anti-E in 6 cases, IgG anti-D 3, IgG anti-cE 1, IgG anti-C 1and IgG anti-c 1. Among them, 3 were seriously ill and treated with blood exchange. 【Conclusion】 As two-child policy was implemented, the incidence of Rh HDN had increased. ABO, RhD, C, c, E and e matched transfusion should be administered for women at childbearing age. Meanwhile, clinical termination of delivery was recommended for pregnant women, who probably develop Rh-HDN and are with critical situation. Rh phenotype matched fresh blood should be prepared, which has great clinical significance for rescuing newborns.

【Objective】 To investigate the distribution characteristics of Rh blood group antigen phenotypes, haplotypes and irregular antibodies between patients in our hospital and local blood donors, so as to ensure safe and effective blood transfusion and improve the rationality and scientificity of clinical blood transfusion. 【Methods】 A total of 113 326 blood samples, from hospitalized patients in our hospital and local blood donors from October 2015 to March 2020, were subjected to Rh antigen typing and irregular antibody detection. The frequency of Rh phenotypes, haplotypes, and irregular antibodies were retrospectively analyzed and calculated. Chi square test was used to compare the data among different population groups. Rh antigen typing and irregular antibody detection were completed using the automatic blood group analyzer. 【Results】 The prevalence of negative RhD was 0.36% (408/113 326). The most prevalent Rh phenotype was DCCee [40.69%(46 112/ 113 326)] followed by DCcEe [36.82%(41 727/ 113 326)]. Anti-E was the most common irregular antibody, accounting for [0.26%(295/ 113 326)], and DCe [62.51%(70 840/ 113 326)] was the most common haplotype. The most common Rh phenotypes and haplotypes in Caucasians in Germany, North Indian and North African were DCcee, DCCee and Dccee, while DCe, DCe and Dce, respectively. 【Conclusion】 The distribution characteristics of Rh phenotypes, haplotypes and irregular antibodies of patients in our hospital and local blood donors were in line with the distribution characteristics of the population in northern China. Corresponding plans concerning blood storage and collection, as well as the establishment of Rh blood type registry should be carried to effectively ensure the safety, rationality and accuracy of clinical blood transfusion.

Objective To evaluate the effect of dexmedetomidine on the postoperative pulmonary function in patients undergoing laparoscopic operation.Methods Eighty American Society of Anesthesiologists physical status Ⅰ or Ⅱ patients,aged 45-64 yr,weighing 45-70 kg,undergoing laparoscopic radical resection of rectal cancer,were divided into 2 groups (n =40 each) using a random number table:control group (group C) and dexmedetomidine group (group Dex).General anesthesia combined with epidural anesthesia was used.In group Dex,dexmedetonidine was intravenously infused as a bolus of 0.3 μg/kg over 10 min after epidural catheterization and before induction of general anesthesia,followed by an infusion of O.4 μg · kg-1 · h-1 starting from the end of tracheal intubation until 30 min before the end of operation.The equal volume of normal saline was given instead in group C.After epidural catheterization and before infusion of dexmedetomidine (T0),immediately before termination of pneumoperitoneum (T1) and at 1,6 and 24 h after operation (T2-4),blood samples were taken from the peripheral vein for determination of concentrations of serum interleukin-6 (IL-6),IL-10,tumor necrosis factor-alpha and malondialdehyde,and arterial blood gas analysis was performed simultaneously.Respiratory index and oxygenation index (OI) were calculated,and the occurrence of OI ≤ 300 mmHg was recorded.Results Compared with group C,the serum IL-6 and tumor necrosis factor-alpha concentrations and respiratory index were significantly decreased,and IL-1O concentrations and OI were increased at T1-4,and malondialdehyde concentrations were decreased at T1-3,and the incidence of OI ≤ 300 mmHg was decreased in group Dex (P＜0.05).Conclusion Dexmedetomidine can inhibit inflammatory responses and improve the postoperative pulmonary function in patients undergoing laparoscopic operation.

ObjectiveTo investigate the effect of dexmedetomidine on postoperative intracranial pressure (ICP) in patients with severe brain injury.MethodsNinety ASA Ⅱ or Ⅲ patients with severe brain injury,aged 19-64 yr,with Glasgow coma scale 3-7,undergoing emergency craniotomy,were randomly divided into 3 groups (n =30 each):control group (group C) and 2 different doses of dexmedetomidine groups (groups D1,D2 ).Anesthesia was induced with propofol 1.5-2.0 mg/kg,fentanyl 4 μg/kg and vecuronium 0.1 mg/kg and maintained with iv infusion of propofol and remifentanil and intermittent iv boluses of vecuronium.The patients were mechanically ventilated after tracheal intubation.Dexmedetomidine 0.3 and 0.7 μg· kg- 1· h- 1 were infused after tracheal intubation in groups D1 and D2 respectively and the infusion was maintained for 48 h.ICP was monitored after operation and maintained ＜ 30 mm Hg by iv injection of hexadecadrol,mannitol and glycerol fructose within 2 d after operation.Venous blood samples were obtained from peripheral vein after anesthesia induction and at 6,12 and 24 h after operation (T0-3) for determination of sertum IL-1β and TNF-α concentrations by ELISA.The clinical results were evaluated using Glasgow prognosis score at 3 months after operation.The amount of hexadecadrol,mannitol and glycerol fructose consumed during 2 d after operation was recorded.ResultsCompared with T0,serum IL-1βand TNF-α concentrations were significantly higher at T1-3 in the three groups ( P ＜ 0.05).The serum IL-1β and TNF-α concentrations at T1-3 and the amount of hexadecadrol,mannitol and glycerol fructose consumed were significantly lower,and the clinical results were significant better in groups D1 and D2 than in group C,and in group D2 than in group D1 ( P ＜ 0.05).ConclusionDexmedetomidine can reduce postoperative ICP in patients with severe brain injury in a dose-dependent manner which is helpful for improving prognosis.The decrease in IL-1β and TNF-α levels and inhibition of inflammatory response may be involved in the mechanism.

Objective To investigate the effect of preemptive analgesia with parecoxib sodium on postoperative cognitive junction in the elderly patients.Methods Sixty ASA ⅠorⅡ patients aged 69-83 yr weighing 47-73 kg undergoing elective operation on the femoral head or hip joint were randomly divided into 2 groups (n=30 each):control group (group C) and parecoxib group (group P).Their preoperative mini-mental state examination scores were more than 23.Parecoxib sodium 40 mg in 5 ml normal saline was injected iv before induction of anesthesia in group P. Patient-controlled intravenous analgesia with fentanyl and tramadol was used in all patients. VAS score was maintained ≤3. If VAS score was more than 3, a bolus of fentanyl 0.2 μg/kg was given iv.Venous blood samples were taken at 1 day before and 1, 4, 24 and 72 h after operation for determination of plasma concentrations of cortisol.Cognitive function was assessed by mini-mental state examination. The total amount of fentanyl and tramadol consumed was recorded. Results Plasma cortisol concentrations and incidence of postoperative cognitive dysfunction were significantly lower and the total amount of fentanyl and tramadol consumed was smaller in group P than in group C.Conclusion Preemptive analgesia with parecoxib sodium 40 mg can reduce the incidence of postoperative cognitive dysfunction in the elderly patients.

Objective To evaluate the incidence of postanesthetic shivering in patients undergoing radical operation for lung cancer under dexmedetomidine-assisted anesthesia.Methods Eighty ASA Ⅰ or Ⅱ patients,aged 50-75 yr,height 158-180 cm,weighing 48-79 kg,scheduled for radical operation for lung cancer under general anesthesia combined with epidural anesthesia,were randomly divided into 2 groups ( n =40 each): normal saline control group (group C) and dexmedetomidine group(group D).Anesthesia was induced with midazolam,fentanyl,propofol and rocuronium.The patients were tracheal intubated and mechanically ventilated.Anesthesia was maintained with sevoflurane inhalation and intermittent epidural ropivacaine.BIS was maintained at 40-55.Group D received dexmedetomidine intravenous infusion at 0.5 μg · kg- 1· h- 1 from immediately after tracheal intubation until 1 h before the end of operation,and group C received the equal volume of normal saline.The shivering within 1 h after operation was scored and recorded.Results The incidence rate of postanesthetic shivering was 50％ in group C and 10％ in group D.Compared with group C,the incidence rate and degree of shivering were decreased in group D( P ＜ 0.05).Conclusion Dexmedetomidine-assisted anesthesia can reduce the incidence of postanesthetic shivering in patients undergoing radical operation for lung cancer.