Objective To explore the optimal method for preparing fresh and fixed skeletal muscle tissues,and to lay an experimental foundation for the rapid diagnosis of and research into the pathogenesis of skeletal muscle diseases.Methods The tibialis anterior muscle was extracted from C57BL/6J mice.Fresh tissue was treated by direct rapid freezing in liquid nitrogen,embedding combined with liquid nitrogen freezing,and foreign body alkane treatment combined with liquid nitrogen freezing.Fixed tissues were pre-treated by direct embedding with embedding agent combined with rapid liquid nitrogen freezing.The frozen sections were stained with hematoxylin and eosin.The cross-sectional areas of ice crystals and muscle fibers were calculated to evaluate the effects of the different pre-treatment method.Results The morphology of the muscle fiber bundles was disrupted and numerous ice crystal vacuoles were observed in fresh tissues after direct liquid nitrogen freezing and foreign body alkane treatment combined with liquid nitrogen freezing.In contrast,the muscle fiber bundles were intact and dense and there were no ice crystals in tissues treated with embedding agent combined with rapid liquid nitrogen freezing,indicating that this pre-treatment method was suitable for preparing fresh skeletal muscle tissue.Fixed tissue treated with embedding agent and liquid nitrogen freezing also showed complete muscle fiber bundles and no ice crystals.Conclusions Treatment of fresh and fixed skeletal muscle tissues with embedding agent combined with rapid liquid nitrogen freezing preserves muscle fiber bundles,with no ice crystals.Tissues prepared by this method are thus suitable for further examinations,such as immunohistochemistry and immunofluorescence.This method will therefore aid the accurate and rapid diagnosis of and research into the pathogenesis of skeletal muscle diseases.

Objective:To investigate the clinical effect and safety of camrelizumab combined with induction chemotherapy followed by concurrent chemoradiotherapy for patients with locally advanced nasopharyngeal carcinoma (NPC).Methods:A total of 24 patients with stage Ⅲ-IV A NPC were recruited prospectively to receive two cycles of camrelizumab combined with induction chemotherapy (docetaxel 75 mg/m 2+ cisplatin 25 mg/m 2 for three consecutive days) followed by concurrent chemoradiotherapy (prescription doses: 6 996 cGy in 33 fractions for PGTV and PGTV nd, 6 006 cGy in 33 fractions for PTV 1, 5 096 cGy in 28 fractions for PTV 2, and concurrent cisplatin chemotherapy with a dose of 75 mg/m 2). The short-term efficacy and adverse reactions were evaluated. Results:After induction therapy, nasopharyngeal lesions showed an objective response rate (ORR) of 91.6%, including 45.8% of complete response (CR) and 45.8% of partial response (PR); cervical lymph nodes showed an ORR of 95.8% (CR: 4.2%; PR: 91.6%). Seventeen patients accepted a reexamination under a nasopharyngoscope, and the biting biopsy result indicated that 13 patients among them had complete pathologic response. After concurrent chemoradiotherapy, nasopharyngeal lesions and cervical lymph nodes showed CR rates of 83.3% and 91.7% and PR rates of 16.7% and 8.3%, respectively. After the induction therapy, 13 patients with stage IV A NPC had ORR (PR) rates of 92.4% and 92.4%, respectively, at nasopharyngeal lesions and cervical lymph nodes. After concurrent chemoradiotherapy, the patients with stage IV A NPC had CR rates of 84.6% and 92.3% and PR rates of 15.4% and 7.7%, respectively, at nasopharyngeal lesions and cervical lymph nodes. Major adverse reactions include leukopenia, granulopenia, anemia, radioactive acute oropharyngeal mucositis and dermatitis, digestive tract reaction, fatigue, hypothyroidism, aminotransferase elevation, and reactive capillary hyperplasia. Conclusions:Camrelizumab combined with induction chemotherapy followed by concurrent chemoradiotherapy can achieve high short-term efficacy for patients with locally advanced nasopharyngeal carcinoma, without increasing the incidence of adverse reactions. Its long-term efficacy deserves further research.

Objective:To investigate imaging characteristics of papulopustular rosacea (PPR) by high-frequency ultrasound combined with color Doppler flow imaging.Methods:From August 2021 to August 2022, 30 patients with PPR were enrolled from the Department of Dermatology, the First Affiliated Hospital of Baotou Medical College in the Inner Mongolia Autonomous Region, and 30 healthy volunteers served as controls. The 22-MHz high-frequency ultrasound combined with color Doppler blood flow imaging was performed to measure the skin thickness, echo and blood flow parameters at the cheek, and the ultrasound results were compared between the two groups. Comparisons between groups were conducted by using t test or chi-square test. The diagnostic value was analyzed using the area under the curve (AUC) in the receiver operating characteristic (ROC) curve. Results:In the case group, there were 12 males and 18 females, and their ages ranged from 22 to 65 years (42.3 ± 12.8 years) ; in the control group, there were 10 males and 20 females, and their ages ranged from 24 to 62 years (41.0 ± 8.4 years) . The epidermal and dermal thicknesses at the cheek were significantly higher in the case group (132.64 ± 12.29 μm, 1 812.29 ± 85.52 μm, respectively) than in the control group (104.34 ± 14.45 μm, 1 671.77 ± 146.55 μm, respectively, both P < 0.05) . High-frequency ultrasound images showed that the case group was mainly characterized by irregular hypoechoic areas in the cheek dermis (80%) , while banded moderately echoic areas were common in the cheek dermis in the control group (90%) ; subepidermal low-echogenic bands and dermal irregular hypoechoic areas were more likely to appear in the case group than in the control group (93.33% vs. 43.33%, 80% vs. 10%, respectively, both P < 0.001) . Compared with the control group, the case group showed a significantly increased proportion of patients with abundant blood flow signals (93.3% vs. 10%, P < 0.05) , and significantly increased blood vessel diameters (1.60 ± 0.42 mm vs. 0.95 ± 0.32 mm, P < 0.05) ; there was no significant difference in peak systolic blood flow velocity and vascular resistance index between the two groups (both P > 0.05) . The AUC of high-frequency ultrasound combined with color Doppler flow imaging quantitative parameters (including epidermal thicknesses, dermal thicknesses, and blood vessel diameters) was 0.989 (95% CI: 0.970 - 1.000) for the diagnosis of PPR, and the sensitivity and specificity were both 96.7%, which were higher than those of single parameter-based diagnostic model. Conclusion:High-frequency ultrasound combined with color Doppler flow imaging can help improve the accuracy of the diagnosis of PPR, by accurately and non-invasively measuring skin thickness and blood flow parameters.

ObjectiveTo systematically analyze the effect of therapeutic exercise on neck function and quality of life in patients with neck pain and forward head posture. MethodsRandomized controlled trials about the effects of exercise training on forward head posture and neck pain were searched from PubMed, Web of science, Embase, Medline, Science Direct, EBSCO, Springlink, CNKI, VIP, and Wanfang Data from database establishment to April, 2022. The literature was screened by two researchers independently. Cochrane bias risk assessment tool and Physiotherapy Evidence Database Scale were used to evaluate the quality of the included articles. Revman 5.4 software was used for meta-analysis. ResultsA total of 416 patients from eleven literatures were included. Level 1a evidence indicated scapula stability training could effectively improve cranial vertebral angle (MD = 3.62, 95%CI 2.41 to 4.83, P < 0.001), and relieve pain (MD = 1.32, 95%CI 0.18 to 2.46, P = 0.02). Level 1b evidence indicated scapula stability training could reduce functional disability (MD = -0.92, 95%CI -1.11 to -0.74, P < 0.001). Level 1b evidence indicated deep cervical flexor training could improve cranial vertebral angle (MD = -0.83, 95%CI -1.56 to -0.10, P = 0.03), relieve pain (MD = 0.93, 95%CI 0.54 to 1.32, P < 0.001), and improve neck functional disability (MD = 2.17, 95%CI 1.39 to 2.95, P < 0.001). ConclusionScapula stability training and deep cervical flexor training can effectively improve cranial vertebral angle, relieve neck pain, and improve neck function.

Objective:To examine the burden of caregivers for elderly Parkinson's patients(PD)and to analyze the mediating effects of social support on primary PD caregivers' burden and their quality of life.Methods:In this cross-sectional study, 281 primary caregivers for elderly Parkinson's patients who visited the Department of Neurology's outpatient clinics of Qilu Hospital of Shandong University from January to May 2021 were surveyed via a convenience sampling method, with 242 valid questionnaires(86.1%). The Chinese versions of the Caregiver Burden Questionnaire, Social Support Rating Scale and 36-Item Short Form Health Survey were used for the investigation.Pearson correlation analysis was used to establish a structural equation model for the influence of social support in the burden and quality of life of caregivers for PD patients.The mediating effects were tested using Process v2.16.3 and re-tested using AMOS.The mediating effects were validated with Bootstrap.Results:Primary caregivers' scores on burden, social support and quality of life were(26.9±23.1), (40.5±8.7)and(593.7±163.3), respectively.Caregiver burden was negatively correlated with social support(r=-0.482, P<0.01), and social support was positively correlated with quality of life(r=0.513, P<0.01). Caregiver burden was negatively correlated with quality of life( r=-0.654, P<0.01), The validated mediating effects showed caregiver burden's direct effects on quality of life(95% CI: -0.38-0.61)and mediating effects of social support(95% CI: -0.08-0.21), not including 0, indicating that the care burden of primary caregivers for elderly PD patients was not only able to predict their quality of life, but also predict their quality of life through the mediating effects of social support(direct effect: -0.50 and mediating effect: -0.14, accounting for 78.0% and 22.0%, respectively, of the total effect of -0.64). Conclusions:Care burden and social support are important factors affecting the quality of life of primary caregivers for elderly PD patients.Reducing care burden can not only directly affect the quality of life, but also indirectly affect the quality of life by increasing social support.

ObjectiveTo explore the effect of exercise on the postural control and muscular function around the ankle in patients with functional ankle instability (FAI) with meta-analysis. MethodsRandomized controlled trials about therapeutic exercise for FAI published before December, 2021 were searched from PubMed, EBSCO, SPORTdiscus, Medline, Science Direct, Springlink, Web of Science, Embase CNKI, VIP and Wanfang Data. The quality and evidence grades of the researches were evaluated by two researchers, and the outcomes were analyzed with RevMan 5.4. ResultsFourteen randomized controlled trials were finally included, involving 434 subjects. Compared with no exercise intervention, therapeutic exercise might significantly improve the movement of center of pressure whether with eye-open or eye-closed (eye-open, SMD = -0.28, 95%CI -0.46 to -0.09, P = 0.003; eye-closed, SMD = -0.24, 95%CI -0.40 to -0.09, P = 0.001); while therapeutic exercise might also enhance the activation of the peroneus longus before dynamic task (SMD = 0.38, 95%CI 0.05 to 0.71, P = 0.03), and activation of the peroneus longus (SMD = 0.53, 95%CI 0.16 to 0.90, P = 0.005) and tibialis anterior (SMD = 0.47, 95%CI 0.10 to 0.84, P = 0.01) after dynamic task. There was neither significant difference in the activation of the tibialis anterior (SMD = 0.48, 95%CI -0.14 to 1.11, P = 0.13), nor the peak torque ratio of eversion to inversion isokinetic strength (SMD = -0.15, 95%CI -0.46 to 0.16, P = 0.340) before dynamic task between the two groups. ConclusionTherapeutic exercise can decrease movement of center of pressure, enable anticipatory contraction of peroneus longus before dynamic tasks and compensatory contraction of peroneus longus and tibialis anterior after tasks, to make it easier to deal with external interference, maintain articular stability and prevent re-injury.

BACKGROUND: RIPA Buffer exhibits different extraction efficiencies of proteins of cells and tissues, which is not appropriate for all samples. OBJECTIVE: To achieve an optimal lysis buffer for skeletal muscle protein extraction in mice of acidosis, and to provide basis for studies on skeletal muscle atrophy. METHEDS: Twenty male healthy C57BL/6 mice, aged 3 months, weighting 25-30 g, were provided by Laboratory Animal Center of Shanxi Medical University. The mice were sacrificed after anesthesia, and the gastrocnemius muscle of lower extremity was isolated. There were two groups: acidosis group was given 10 g of feed mixed with 0.4 mol/L hydrochloric acid (10 mL) , and control group received 10 g of feed mixed with same volume of water, for 7 consecutive days. The effect of RIPA Buffer, Original Buffer and JP Buffer on the skeletal muscle protein extraction in mice of acidosis was compared. The expression levels of AKT, p-AKT (Thr308) , rpS6 and p-rpS6 (Ser235/236) were detected by western blot assay. GLUT4 mRNA expression was examined by RT-qPCR. RESULTS AND CONCLUSION: (1) Different buffers generated different protein-yields. The protein yield was highest in JP Buffer, but the target protein signal was not high. The protein yield was low in RIPA Buffer. Original Buffer could extract sufficient proteins, and had clear band detected by western blot assay. (2) Western blot assay scores in Original Buffer were higher than those of other two buffers. (3) Western blot assay results showed that the extent of phosphorylation in both groups showed no significant changes. (4) GLUT4 mRNA expression level examined by RT-qPCR showed no significant changes in both groups. (5) These results indicate that Original Buffer is optimal lysate of skeletal muscle protein extraction. Inactivated AKT signaling pathway is seen in the short-term hydrochloric acid-induced acidosis group, so whether lengthening acidosis time can activate the signaling pathway. Selecting the optimal lysis buffer for different samples is premise to ensure western blot assay results.

Objective To investigate the effect of tripterygium glycosides combined with recombinant human tumor necrosis factor receptor Ⅱ antibody fusion protein for injection on serum vascular endothelial growth factor (VEGF),rheumatoid factor (RF),receptor activator of nuclear factor κ B ligand (RANKL) levels in patients with rheumatoid arthritis.To provide reference for rational clinical application.Methods From December 2014 to January 2016,132 patients with rheumatoid arthritis were divided into observation group and control group by random number table method,,with 66 cases in each group,The control group was treated with tripterygium glycosides alone (10 mg each time,3 times daily,orally) for 3 months.The observation group was treated with a combination of tripterygium glycosides and recombinant human tumor necrosis factor receptor Ⅱ (0.4 mg/kg,once weekly,hypodermic injection) for 3 months.The clinical efficacy,and serum VEGF,RF and RANKL levels were compared between 2 groups.Results The effective rate of the observation group was 92.4％ (61/66),which was significantly higher than that in the control group (80.3％,53/66).There was a significant difference between the two groups (x2=4.117,P＜0.05).There was no significant difference in the levels of serum VEGF,RF and RANKL between the two groups (t =0.174,0.103,0.359,all P＞0.05).After treatment,the levels of serum VEGF,RF and RANKL in the observation group and the control group were (20.8± 11.5) ng/L and (27.3 ±13.1) ng/L,(258.4±54.5) U/L and (298.1 ±49.5) U/L,(0.083±0.021) pmol/L and (0.197 ± ±0.064),respectively.There were significant differences between the two groups (t =3.029,4.381,13.750,all P＜0.05).The incidence rate of adverse reactions in the control group was 10.5％ (7/66),which was significantly higher than that in the observation group (1.5％,1/66).There was a significant difference between the two groups(x2 =4.790,P＜0.05).The one-year recurrence rate was 25.0％ (13/52) in the control group and that was 6.7％ (4/60) in the observation group,respectively,and there was a significant difference between the two groups (x2 =7.272,P＜ 0.05).Conclusion Tripterygium glycosidescombined with recombinant human tumor necrosis factor receptor Ⅱ antibody fusion protein for injection is effective in the treatment of rheumatoid arthritis,which reduces the levels of serum VEGF,RF and RANKL,and has a low incidence of adverse reactions and recurrence.

Objective To explore the effects of dietary approaches to stop hypertension (DASH) on the quality of life and re-hospitalization rate in patients with chronic heart failure (CHF). Methods A total of 163 patients with CHF receiving standard medical therapy in the Cardiology Department of the First Affiliated Hospital of Zhengzhou University from February to September 2014 were selected and divided into experimental group (n=85) and control group (n=78) randomly. Low salt, low fat diet was applied in the control group, while DASH was adopted in the experimental group. Patients' body mess index (BMI), blood pressure, blood glucose, blood lipid, ejection fraction, cardiac function, the score of Minnesota Living with Heart Failure Questionnaire (MLHFQ) and re-hospitalization rate were compared before and after intervention. Results There were no statistical differences in BMI, blood pressure, blood glucose, blood lipid, cardiac function and the score of MLHFQ before intervention (P> 0.05). Patients' BMI, systolic pressure, cholesterol and triglyceride of the experimental group paitents were all lower than those of the control group 12 months after intervention, with statistical significance (P< 0.05). 12 months after intervention, patients' quality of life of the experimental group was higher than that of the control group (P<0.05), and the re-hospitalization rate was lower than that of the control group (P<0.05). Conclusions DASH can improve the quality of life in patients with CHF, reduce readmission rate, thus improving the prognosis of heart failure.

OBJECTIVETo monitor human cytomegalovirus (HCMV) drug resistance in recipients of hematopoietic stem cell transplantation by phenotypic and genotypic methods.

METHODSHCMV clinical isolates was isolated from the urine of hematopoietic stem cell transplantation recipients treated with GCV. Tissue cell infection median dose (TCID50) of the isolates was calculated using Reed-Muench method, and their drug susceptibility was determined by plaque reduction assay. We amplified the UL97 DNA fragment of the virus by nested PCR followed by automated DNA sequencing.

RESULTSHCMV clinical strain isolated from the urine samples of the recipients using a human fibroblast cell line showed a TCID50 value of 10(-4.618)/0.1 ml and a 50% inhibitory concentration (IC50) to GCV of 5.847 µmol/L, suggesting its sensitivity to GCV. Alignment with the AD169 DNA reference sequence identified 4 point mutations of the virus at 1509 (T-C), 1575 (C-T), 1794 (T-C), and 1815 (C-G), and only the last mutation resulted in one amino acid mutation to D605E. No gene mutation was found in relation to GCV resistance.

CONCLUSIONSPhenotypic and genotypic assays were established to examine antiviral drug resistance of HCMV in recipients of hematopoietic stem cell transplantation. We did not find any drug resistance of the clinical HCMV isolate.

Antiviral Agents ; pharmacology ; Cell Line ; Cytomegalovirus ; drug effects ; genetics ; isolation & purification ; Drug Resistance, Viral ; genetics ; Ganciclovir ; pharmacology ; Genes, Viral ; Genotype ; Hematopoietic Stem Cell Transplantation ; Humans ; Mutation ; Phosphotransferases (Alcohol Group Acceptor) ; genetics