OBJECTIVE To study the chemical constituents in S henfu injection and the anti-inflammatory activities of its polyacetylene compounds. METHODS Shenfu injection was separated and purified by macroporous adsorption resin ，medium pressure liquid chromatography ，preparative thin layer chromatography and reversed-phase semi-preparative high-performance liquid chromatography，and the compound structure was identified according to the physicochemical properties and spectral data. RAW 264.7 cell inflammation model was used to evaluate the anti-inflammatory activities of polyacetylene compounds . The effects of active polyacetylene compounds on the expressions of cyclooxygenase- 2（COX-2）protein were evaluated by Western blot assay. RESULTS Twelves compounds were isolated and identified from Shenfu injection ，including 8 ginsenoside compounds ，i.e. ginsenoside Rg 1（1），ginsenoside Re （2），ginsenoside Rb 1（3），ginsenoside Rk 1（4），20（R）-ginsenoside Rh 1（5），20（S）-ginsenoside Rg3 （6），notoginsenoside R 1（7），panaxatriol（8）；4 polyacetylene compounds ，i.e.（3R，9R，10R）-panaxytriol（9），panaxydol（10）， heptadeca-1，8-dien-4，6-diyne-3，10-diol（11）and panaxynol （12）. Among 4 polyacetylene compounds ，only compound 10 had anti-inflammatory activity. Compound 10 was not toxic to normal RAW 264.7 cells；when the concentration of compound 10 ranged 12.5-50.0 μmol/L，it could significantly reverse the lipopolysaccharide-induced NO content increase in cell supernatant （P＜0.05 or P＜0.01）；when the concentration of co mpound 10 was 50.0 μmol/L，it could significantly reverse the lipopolysaccharide-induced protein expression increase of COX- 2 in cells （P＜0.05）. CONCLUSIONS Compounds 4，7，10-12 are identified and reported in Shenfu injection for the first time ，and panaxydol possesses a certain anti-inflammatory effect.

Objective:To investigate the molecular mechanism of rabbit ear cartilage remodeling induced by 1064 nm Nd∶YAG laser irradiation.Methods:Thirty-three rabbits were divided into 6 groups according to a random number table, and were used in the following experiments respectively: screening the optimal laser energy (6 rabbits), observing the changes of ear cartilage tissue immediately (6 rabbits), 1 week (6 rabbits), 3 weeks (6 rabbits), and 6 weeks after irridiation (6 rabbits), as well as transcriptome sequencing and verification (3 rabbits). The left ears of rabbits were used as untreated self-controls, while the right ears were irradiated. (1) Laser energy screening: the right ear cartilages of 6 rabbits were irradiated with 1064 nm Nd∶YAG laser at different energy densities (50, 60, 70, 80, 90, 100 J/cm 2). Cartilages without skin and perichondrium were cut into the same thickness and size for HE staining, in order to observe the changes of chondrocytes and to determine the optimal laser energy density for shaping (hereinafter referred to as the optimal energy density). (2) Histological observation of rabbit ear cartilage: the right ears of 24 rabbits were irradiated with laser under the optimal energy density. Bilateral cartilage samples were collected immediately after surgery and at 1, 3 and 6 weeks after surgery for histological observation (HE, Masson and Sirius red staining). (3) Transcriptome sequencing and sample verification: the right ears of 3 rabbits were irradiated with laser under the optimal energy density, and the cartilages were harvested and mixed within 6 hours after irradiation, which was set as the laser irradiation group. Cartilages of the same region and size on the left ears were set as blank control group. Transcriptome RNA sequencing was performed, and the expression levels of related genes and proteins were detected by quantitative polymerase chain reaction (qPCR) and Western blotting. Results:(1) After laser irradiation with different energy density of 50-100 J/cm 2, HE staining showed that the chondrocytes could be changed at the laser energy density of 80 J/cm 2, without causing vacuolar deformation and coagulation necrosis, indicating that the damage degree was suitable.(2) After 80 J/cm 2 laser irradiation, histological observation revealed that there was an irradiated zone immediately after the operation. The overall morphology of chondrocytes in the radiation zone was elongated and exhibited spindle cell-like changes, with deep matrix staining and obvious refraction, as well as a relative increase of type Ⅱ collagen. At 1 week after irradiation, the radiation zone became shallower and the cell size recovered. From 3 weeks to 6 weeks, the matrix staining around the radiation zone gradually deepened and the overall cellular morphology was stretched again. (3) Transcriptome RNA sequencing revealed that there were 198 differentially expressed genes (70 up-regulated and 128 down-regulated) in the laser irradiated group compared with the blank control group. Through further screening and study, CREB3L2 gene expression was up-regulated in the laser irradiation group. qPCR results showed that the relative expression level of CREB3L2 mRNA in laser irradiation group was significantly higher than that in the blank control group, the difference was statistically significant( P<0.01). Western blotting showed that the relative expression level of CREB3L2 protein was higher than that of control group in a time-dependent manner, and the difference was statistically significant( P<0.01). Conclusions:After irradiation with long-pulsed 1064 nm Nd∶YAG laser, the expression level of CREB3L2 gene is up-regulated, which then induced chondrocyte rearrangement and proliferation, resulting in morphological and biomechanical changes of ear cartilage.

Objective:To investigate the molecular mechanism of rabbit ear cartilage remodeling induced by 1064 nm Nd∶YAG laser irradiation.Methods:Thirty-three rabbits were divided into 6 groups according to a random number table, and were used in the following experiments respectively: screening the optimal laser energy (6 rabbits), observing the changes of ear cartilage tissue immediately (6 rabbits), 1 week (6 rabbits), 3 weeks (6 rabbits), and 6 weeks after irridiation (6 rabbits), as well as transcriptome sequencing and verification (3 rabbits). The left ears of rabbits were used as untreated self-controls, while the right ears were irradiated. (1) Laser energy screening: the right ear cartilages of 6 rabbits were irradiated with 1064 nm Nd∶YAG laser at different energy densities (50, 60, 70, 80, 90, 100 J/cm 2). Cartilages without skin and perichondrium were cut into the same thickness and size for HE staining, in order to observe the changes of chondrocytes and to determine the optimal laser energy density for shaping (hereinafter referred to as the optimal energy density). (2) Histological observation of rabbit ear cartilage: the right ears of 24 rabbits were irradiated with laser under the optimal energy density. Bilateral cartilage samples were collected immediately after surgery and at 1, 3 and 6 weeks after surgery for histological observation (HE, Masson and Sirius red staining). (3) Transcriptome sequencing and sample verification: the right ears of 3 rabbits were irradiated with laser under the optimal energy density, and the cartilages were harvested and mixed within 6 hours after irradiation, which was set as the laser irradiation group. Cartilages of the same region and size on the left ears were set as blank control group. Transcriptome RNA sequencing was performed, and the expression levels of related genes and proteins were detected by quantitative polymerase chain reaction (qPCR) and Western blotting. Results:(1) After laser irradiation with different energy density of 50-100 J/cm 2, HE staining showed that the chondrocytes could be changed at the laser energy density of 80 J/cm 2, without causing vacuolar deformation and coagulation necrosis, indicating that the damage degree was suitable.(2) After 80 J/cm 2 laser irradiation, histological observation revealed that there was an irradiated zone immediately after the operation. The overall morphology of chondrocytes in the radiation zone was elongated and exhibited spindle cell-like changes, with deep matrix staining and obvious refraction, as well as a relative increase of type Ⅱ collagen. At 1 week after irradiation, the radiation zone became shallower and the cell size recovered. From 3 weeks to 6 weeks, the matrix staining around the radiation zone gradually deepened and the overall cellular morphology was stretched again. (3) Transcriptome RNA sequencing revealed that there were 198 differentially expressed genes (70 up-regulated and 128 down-regulated) in the laser irradiated group compared with the blank control group. Through further screening and study, CREB3L2 gene expression was up-regulated in the laser irradiation group. qPCR results showed that the relative expression level of CREB3L2 mRNA in laser irradiation group was significantly higher than that in the blank control group, the difference was statistically significant( P<0.01). Western blotting showed that the relative expression level of CREB3L2 protein was higher than that of control group in a time-dependent manner, and the difference was statistically significant( P<0.01). Conclusions:After irradiation with long-pulsed 1064 nm Nd∶YAG laser, the expression level of CREB3L2 gene is up-regulated, which then induced chondrocyte rearrangement and proliferation, resulting in morphological and biomechanical changes of ear cartilage.

Objective:To investigate the effects of perioperative whole course thermal insulation strategy on extubation, recovery time, coagulation index and the incidence of complications in patients undergoing thoracoscopic surgery.Methods:A total of 121 patients who underwent thoracoscopic surgery who received treatment in Zhoushan Hospital from October 2016 to February 2018 were included in this study. Among them, 59 patients who underwent thoracoscopic surgery from October 2016 to February 2017 were included in the simple thermal insulation group, and 62 patients who underwent thoracoscopic surgery from October 2017 to February 2018 were included in the whole process thermal insulation group. Before and 24 hours and 72 hours after surgery, platelet count, prothrombin time, activated partial thromboplastin time and thrombin time were compared between the two groups. Postoperative extubation time and recovery time were recorded. The incidence of shivering, restlessness and other complications was analyzed.Results:Postoperative extubation time and recovery time in the whole process thermal insulation group were significantly shorter than those in the simple thermal insulation group [(8.06 ± 4.60) min vs. (13.98 ± 7.22) min, (47.19 ± 12.97) min vs. (56.84 ± 17.49) min, t = 5.40, 3.47, both P < 0.05). At 24 and 72 hours after surgery, platelet count and activated partial prothrombin time in the whole process thermal insulation group were significantly shorter than those in the simple thermal insulation group [(12.55 ± 0.88) s vs. (13.11 ± 0.97) s, (27.44 ± 2.43) s vs. (29.03 ± 2.14) s, (11.42 ± 0.73) s vs. (11.87 ± 0.74) s, (27.44 ± 1.96) s vs. (28.80 ± 2.22) s, t = 3.32, 3.81, all P < 0.05). The incidence of postoperative shivering and restlessness in the whole process thermal insulation group was significantly lower than that in the simple insulation Group (7 cases vs. 27 cases, 5 cases vs. 22 cases, χ2 = 17.782, 14.894, 3.33, 3.57, all P < 0.05). Conclusion:Perioperative whole course thermal insulation strategy can shorten extubation and recovery time, inhibit perioperative coagulation dysfunction, and decrease the incidence of perioperative restlessness, shivering and other complications caused by hypothermia. The effect of perioperative whole course thermal insulation is superior to that of simple thermal insulation.

OBJECTIVE：To establish an HPLC fing erprint of Ginsen g Radix et Rhizoma Rubra ，and to optimize its processing technology. METHODS ：HPLC method was adopted. The determination was performed on Waters SymmetryShield TM RP18 column with mobile phase consisted of acetonitrile-water （gradient elution ）at the flow rate of 1.0 mL/min. The column temperature was set at 30 ℃，and the detection wavelength was 203 nm. The sample size was 10 μL. Using ginsenoside Rb1 as reference peak ，HPLC fingerprints of 10 batches of Ginseng Radix et Rhizoma Rubra was established. The similarity of them was evaluated by using Similarity Evaluation System of TCM Chromatogram Fingerprint （2012 A edition ） to confirm common peak. With steaming temperature，time and drying method as factors ，using the content of ginsenoside and fingerprint similarity as index ，the processing technology was optimized with L 16（43）orthogonal test design and verified. Cluster analysis was conducted with SPSS 19.0 statistical software of 10 batches of Ginseng Radix et Rhizoma Rubra and 3 batches of optimal processed sample. RESULTS ：There were a total of 13 common peaks in the fingerprints of 10 batches of Ginseng Radix et Rhizoma Rubra. The similarity was more than 0.920；3 common peaks were identified ，such as ginsenoside Rg 1，ginsenoside Re ，ginsenoside Rb 1. The optimal processing technology included that steamed at 100 ℃ for 150 min，dried at 60 ℃. The results of validation test show that the contents of ginsenoside Rg 1，Re and Rb 1 were 0.26％-0.29％，0.17％-0.20％，0.47％-0.54％，and the similarity between 3 batches of Ginseng Radix et Rhizome Rubra optimal processed sample and the control fingerprints was more than 0.970. The results of cluster analysis showed that 10 batches of Gimseng Radix et Rhizoma Rubra and 3 batches of optimal processed sample could be clustered into two categories；HS3-HS10 could be clustered into one category ，and 3 batches of optimal processed sample ，HS1 and HS 2 be clustered into one category. CONCLUSIONS ：Established fingerprint can be used for the optimization of processing technology of Gimseng Radix et Rhizoma Rubra ，and characterize the correlation between f luctuation of technology parameter and quality of medicinal material；the optimal processing technology is reasonable an d

Objective To explore the difference of psychological state and the change of psycho logical state between mastectomy group for the purpose of changing gender identity and breast reduction surgery group for the purpose of improving physical beauty.Methods The psychological evaluation scale of three layers and twelve dimensions was used to evaluate the psychological status of 24 patients who underwent clinical breast masculine surgery and aesthetic breast reduction before and after operation.The psychological characteristics between the two groups before and after operation were compared and analyzed.Data analysis was carried out through Stata 13.0 software package.Results There were significant differences in depression factor,anxiety factor and obsessive compulsive symp tom factor in the aesthetic breast reduction group before and after operation (t=2.194,2.183,2.090;P=0.039,0.040,0.048,P＜0.05).There were significant differences in depression factor,anxiety factor,obsessive-compulsive symptom factor,bullying factor,psychological self,physical self and social self before and after mastectomy (t=4.188,3.730,2.484,2.570,5.898,3.531,3.162,P=0.000,0.001,0.021,0.018,0.000,0.002,0.005,P＜0.05).There were significant differ ences in depression,anxiety,psychoticism,obsessive-compulsive symptoms,psychological self,physiological self and social self between mastectomy group and aesthetic mastectomy group before operation.(t=2.629,4.081,2.097,4.246,5.419,2.600,2.545;P=0.015,0.001,0.048,0.000,0.000,0.016,0.018,P＜0.05).There was significant difference in postoperative depression,anxiety and obsessive-compulsive symptoms between the mastectomy group and the aesthetic mastectomygroup (t=2.117,2.170,3.764;P=0.046,0.041,0.001,P＜0.05).Conclusions Plastic surgeons should pay attention to the psychological status and personality characteristics of different beauty seekers and give psychological guidance,which is of great significance to improve the medical experience and surgical effect of the beauty seekers.

Clinical genetic testing results are compiled into a standardized report by genetic specialists and provided to clinicians and patients (Should the patient be intellectually disabled or under 18,the report will be provided to his/her parents or legal guardians).The content of genetic testing report should conform to relevant guidelines,industry standards and consensus.The decisions of clinicians will be made based on the report and clinical indications.Genetic counselors should provide post-test counseling to clinicians and patients or their authorized family members.A mechanism of follow-up visit after the genetic testing should be established with informed consent.Data should be shared by clinical institutions and genome sequencing institutions.As findings upon follow-up visit can help with further evaluation of the results,genome sequencing institutions should regularly re-analyze historical and follow-up data,and the updated results should be shared with clinical institutions.All activities involving reporting,genetic counselling,follow-up visiting,and re-analyzing should follow the relevant guidelines and regulations.

Objective: To investigate the histological and biomechanical changes of rabbit ear cartilage induced by long pulse width 1064 nm Nd: YAGlaser. Methods: Seven New Zealand rabbits weighing 2.0-2.5 kg, male, 4-6 months old were selected. For a self-control, the right ear was irradiated by laser and the left ear was the normal control group. Each rabbit ear was divided into 3 cm × 1 cm, three 2 cm × 1 cm and 1 cm× 1 cm experimental area. After the experimental area of the laser irradiation group was irradiated by long pulse width 1064 nm Nd: YAG laser, Three pieces of 2 cm × 1 cm cartilage in each ear of the experimental rabbits were immediately cut and stained with HE, Sirius red and Masson to observe the histological changes of chondrocytes. The 3 cm × 1 cm and 1 cm× 1 cm cartilage of each rabbit ear was cut for biomechanical examination, and the changes of biomechanical properties such as tension, fracture, elastic modulus and compression of rabbit ear cartilage were observed immediately after the operation. Results: Histological observation showed that the chondrocytes became small, the matrix was stained deeply, the refraction was obvious/evident and the collagen was relatively increased. After being irradiated by long pulse width 1064 nm Nd: YAG laser, the peak tensile stress, tensile elastic modulus, tensile fracture load, compressive stress peak, compressive elastic modulus and compressive failure load of rabbit ear cartilage in laser irradiation group were (5.22 ±0.80) MPa, (42.40 ±9.78) MPa, (22.86 ±4.85) N, (16.04 ±5.57) MPa, (28.71 ±13.97) MPa, (1 211.63 ±427.86) N. All of them were smaller than those of the control group[(6.07±0.64) MPa, (48.44±6.30) MPa, (26.94±4.19) N, (25.12±9.10) MPa, (45.30±19.24) MPa, (1 962.83±896.71) N], and the difference was statistically significant (P<0.05). Conclusions After the operation of long pulse width 1064 nm Nd: YAG laser, the stress of rabbit ear cartilage resisting elastic deformation is reduced, the shape is easier to change, and the remodeling effect is more stable.

Objective: To explore the clinical and molecular genetic features of a Chinese patient with catecholaminergic polymorphic ventricular tachycardia (CPVT). Methods: Clinical data including resting electrocardiography, echocardiography and treadmill exercise testing of a patient with CPVT admitted to our department in March 2013 were analyzed, and the peripheral venous blood samples of the patient and his family members and 400 ethnicity-matched healthy controls were obtained. All exons and exon-intron boundaries of the six CPVT-related genes including RYR2, CASQ2, TRDN, CALM1, KCNJ2 and ANKB were sequenced to detect the variants related to CPVT. The relationship between the genotypes and phenotypes was analyzed to direct the target therapy. Results: Recurrent syncope induced either by exercise or extreme frightened fear was observed in this patient. There was no positive family history of syncope or sudden death. The resting electrocardiography and echocardiography of the patient were normal, while the exercise testing revealed bidirectional and polymorphic ventricular tachycardia. A cardiac ryanodine receptor gene mutation (R2401H) was identified in this patient, while this mutation was absent in his parents and sister and 400 controls. No variant was detected in the remaining five candidate genes. Treatment with high dose of metoprolol succinate (118.75 mg/d) was effective and patient was free of syncopal attack during the 2 years follow-up. Conclusion This is the first report on RyR2-R2401H mutation in Chinese patient with CPVT, and high dose of metoptolol is the effective therapy option for CPVT related to RyR2 mutation.

Objective To establish the methods of microbial limit test for three kinds of preparations with heavy metals, such asDingxian Pills,Pizhi Lotion andJiawei Huangqin Ointment.Methods According to Chinese Pharmacopoeia 2010, the recovery rates of bacteria, fungus and yeast treated by three preparations were detected. And the methods of testing control bacteria were also validated.Results Culture medium dilution method was proved to be applicable forDingxian Pills. Culture medium dilution method combined with pre-filtration method was proper for Pizhi Lotion. And the extraction method was adopted forJiawei Huangqin Ointment. The recovery rates of these five validation strains reached 70% by appropriate methods. And the same methods were used for validation of the control bacteria.Conclusion The methods of microbial limit test for these three different preparations were established through this study.