Objective:A gas chromatography(GC)method was established for the simultaneous determination of seven residual solvents in clopidogrel bisulfate,including methanol,acetone,isopropanol,dichloromethane,n-hexane,tetrahydrofuran and methylbenzene.Methods:The analysis was performed on an Agilent DB-1 capillary column(30 m ×0.32 mm,3.0 μm)under the programmed temperature.The carrier gas was high purity nitrogen,and the flow rate was 4.0 mL·min-1 with constant flow control.The detector was a hydrogen flame ionization detector(FID).The inlet temperature was 200 ℃,the detector temperature was 270 ℃ and the split ratio was 5∶1.The temperature of headspace injector was 85 ℃,and the equilibrium time was 20 min.Results:Seven residual solvents were separated completely.The linear relationship between peak area and the concentration range of seven residual solvents were good enough(r＞0.9993).The results of specificity,precision and recovery met the requirements.Conclusion:This method can be used for the determination of residual organic solvents in clopidogrel bisulfate.

ObjectiveTo identify the functions of the AP2/ERF family members in Pinellia ternata and promote the genetic improvement of P. ternata varieties. MethodWe identified and conducted a systematic bioinformatics analysis of the AP2/ERF family member genes in P. ternata based on the three generations of transcriptome data. Real-time polymerase Chain reaction　（Real-time） PCR was employed to determine the expression pattern of AP2/ERF genes in different tissues and under different stress conditions. ResultA total of eight full-length AP2/ERF family members were identified from the transcriptome data， which were classified into three sub-gene families： AP2， ERF， and DREB. The deduced AP2/ERF proteins in P. ternata had the length of 251-512 aa， the theoretical pI of 5.29-11.72， the instability index of 45.90-82.41， subcellular localization in the nucleus， and conserved domains and motifs. AP2/ERF genes were expressed in different tissues of P. ternata， with high expression levels in the leaf. The stress response experiments showed that PtERF1 mainly responded to NaCl stress. The expression of PtERF2 and PtERF4 was significantly up-regulated under low temperature and polyethylene glycol （PEG）-simulated stress. PtERF3 responded to both low temperature and NaCl stress. The expression of PtERF5 was induced by high temperature， low temperature， NaCl and PEG stress. The expression of PtERF7 was up-regulated under high temperature， while that of PtERF8 under low temperature. ConclusionThe AP2/ERF genes in P. ternata can respond to stress and have the potential functions of regulating photosynthesis and improving root stress resistance.

ObjectiveTo compare the retest reliability and discriminant validity of dynamic postural stability indices for functional ankle instability (FAI) obtained by different algorithms based on acceleration signals at different positions of human body. MethodsFrom April to June, 2021, 21 subjects with unilateral FAI and 21 subjects with normal ankle were recruited. Three inertial sensors were attached to the waist points, knee and ankle positions. The ground reaction force (GRF) and kinematics data of the subjects in multi-direction single leg landing test were collected synchronously by 3D force plate and inertial sensors. The unbounded third order polynomial (UTOP) fitting method was used to calculate the stability time, and the root mean square was used to caculate the stability index. ResultsMost of the indicators calculated based on acceleration signal correlated with that based on GRF with low coefficient (|r| = 0.116 to 0.368, P < 0.05). The stability time and stability index based on the acceleration signals of different positions of human body showed low to high retest reliability (CMC 0.30 to 0.91). For the females, among the stability time based on acceleration signal, eleven indexes achieved average to very high discriminant validity (AUC = 0.702 to 0.942, P < 0.05); eight of the stability indexes reached general level of discriminant validity (AUC = 0.717 to 0.782, P < 0.05). No algorithms achieved good discriminant effect in male subjects. ConclusionBased on the acceleration signal of waist point in single-leg landing stability test, the stability time calculated by UTOP algorithm can evaluate the dynamic postural stability of female FAI patients with high discriminant validity and medium to high retest reliability.

Bipolar androgen therapy (BAT), as a new therapy, can effectively reduce the serum prostate specific antigen (PSA) level of a part of patients with castration resistant prostate cancer (CRPC), delay tumor progression, improve their quality of life and restore the sensitivity to drug therapy. This paper will review the background, possible mechanism, clinical research progress and development prospect of BAT.

Objective To build a method for calculating the optimal length of hamstring muscles in vivo, and make comparison with other indirect parameters which represent the optimal length. Methods By synchronously recording knee flexion torque and kinematic data, the musculoskeletal model of lower limbs was built to obtain hamstring strength and muscle length, and to further calculate the optimal length of hamstring muscles. Results Flexion angle at peak knee flexion torque was significantly greater than that at peak hamstring strength and their correlation coefficient was 0.741. The optimal lengths of each bi-articulated hamstring muscles were significantly greater than the corresponding muscle lengths during standing and their correlation coefficients was low. Conclusions The established estimating method for optimal length of hamstring muscles provided references for future studies on injury mechanism and risk factors. Flexion angle at peak knee flexion torque could partly represent the optimal length of hamstring. It is not suggested that hamstring muscle length during standing should be used as an approximation of hamstring optimal length.

The standardized residency training has become a necessary way to improve the overall professional quality of doctors. At present, a single teaching model can't meet the needs of the standardized residency training. This study explores the application of problem-based learning (PBL) combined with multidisciplinary treatment (MDT) model in the general surgery teaching of standardized residency training, so that the residents can use their theoretical knowledge to think deeply about the problems in the cases. By consulting the literature review, the cases are analyzed from multiple levels and angles, such as pathogenic causes, pathogenesis and clinical manifestations, then the clinic diagnosis and therapeutic schedule can be obtained. This kind of teaching model not only stimulates the group's more interest in learning, and improves the ability of autonomous learning, independent analysis, problem solving and language expression, but also significantly improves teaching satisfaction and has obvious teaching advantages.

Objective:To evaluate the role of protein kinase C-delta (PKC-δ) in ventilator-induced lung injury (VILI) and the relationship with NLR family CARD domain-containing protein 4 (NLRC4) in rats.Methods:Thirty-six clean-grade healthy adult male Sprague-Dawley rats, weighing 200-250 g, were divided into 3 groups ( n=12 each) using a random number table method: control group (group C), VILI group (group V) and VILI plus KAI 9803 group (group VK). In V and VK groups, tracheal tubes were placed for mechanical ventilation after tracheotomy, ventilator settings were adjusted with a tidal volume of 40 ml/kg, respiratory rate of 60 breaths/min, and inspiratory/expiratory ratio of 1∶2, and air was inhaled.Group C received no mechanical ventilation after tracheal intubation.Immediately after completion of intubation, PKC-δ specific inhibitor KAI 9803 200 μg/kg was intratracheally injected in group VK, and the equal volume of phosphate buffer saline was given instead in the other two groups.Blood samples were taken from the femoral artery at 4 h of mechanical ventilation to record PaO 2.The chest was opened at the end of mechanical ventilation, lung tissues were removed, and the left lung tissues were lavaged to collect bronchoalveolar lavage fluid (BALF). The pathological changes of lung tissues were examined with a light microscope and scored.Enzyme-linked immunosorbent assay was used to detect the concentrations of interleukin-1beta (IL-1β) and IL-18 in BALF, Western blot was used to detect the expression of NLRC4, caspase-1 and PKC-δ in the right lower lobe of the lung, and the expression of NLRC4 mRNA in the right lower lobe of the lung was determined by real-time polymerase chain reaction, and the wet/dry weight ratio (W/D ratio) of the right middle lobe of the lung was calculated. Results:Compared with group C, the pathological score and W/D ratio were significantly increased, PaO 2 was decreased, the concentrations of IL-1β and IL-18 in BALF were increased, and the expression of NLRC4, caspase-1 and NLRC4 mRNA was up-regulated in V and VK groups, and the expression of PKC-δ was significantly up-regulated ( P<0.01), and a large amount of edema fluid was seen in the alveolar space, with inflammatory cell infiltration in group V ( P<0.01). Compared with group V, the pathological score and W/D ratio were significantly decreased, PaO 2 was increased, the concentrations of IL-1β and IL-18 in BALF were decreased, the expression of NLRC4, caspase-1, PKC-δ and NLRC4 mRNA was down-regulated ( P<0.05), and fluid exudation in the alveolar space and the degree of inflammatory cell infiltration were significantly attenuated in group VK. Conclusion:PKC-δ is involved in VILI, which is related to inhibiting NLRC4 expression in rats.

Objective:To evaluate the effect of rapamycin on the activity of NOD-like receptor C4 (NLRC4) inflammasomes in the rats with ventilator-induced lung injury (VILI).Methods:Thirty-six healthy clean-grade male Sprague-Dawley rats, aged 6-8 weeks, weighing 200-250 g, were divided into 3 groups ( n=12 each) using a random number table method: control group (group C), VILI group and rapamycin group (group RAPA). In group RAPA, rapamycin 4 mg·kg -1·d -1 was intraperitoneally injected once a day for 3 consecutive days before establishing the model, while the equal volume of normal saline was given instead in group C and group VILI.The patients were mechanically ventilated for 4 h (tidal volume 20 ml/kg, respiratory rate 80 breaths/min, inspiratory/expiratory ratio 1∶1, fraction of inspired oxygen 21%) in VILI and RAPA groups.Blood samples were collected from the femoral artery after the end of ventilation for blood gas analysis and for determination of serum interleukin-1β (IL-1β) and interleukin-18 (IL-18) concentrations (by enzyme-linked immunosorbent assay), and PaO 2 was recorded.The bronchoalveolar lavage fluid (BALF) was collected for determination of the neutrophil count and IL-1β and IL-18 concentrations by enzyme-linked immunosorbent assay.The lung tissues were obtained for examination of the pathological changes (under the light microscope) after HE staining which were scored and for determination of wet to dry weight (W/D) ratio, and expression of mammalian target of rapamycin (mTOR), NLRC4 and caspase-1 (by Western blot) and expression of NLRC4 mRNA (by real-time polymerase chain reaction). Results:Compared with group C, the W/D ratio, lung injury score, neutrophil counts in BALF, and concentrations of IL-1β and IL-18 in serum and BALF were significantly increased, PaO 2 was decreased, and the expression of mTOR, NLRC4, caspase-1 and NLRC4 mRNA was up-regulated in group VILI and group RAPA ( P<0.01). Compared with group VILI, the W/D ratio, lung injury score, neutrophil counts in BALF, and concentrations of IL-1β and IL-18 in serum and BALF were significantly decreased, PaO 2 was increased, and the expression of mTOR, NLRC4, caspase-1 and NLRC4 mRNA was down-regulated in group RAPA ( P<0.05). Conclusion:The mechanism by which rapamycin alleviates VILI may be related to inhibiting activation of mTOR signaling pathway and inhibiting the activity of NLRC4 inflammasomes in rats.

Objective To investigate the effects of the polymorphisms in the promoter of ATP binding cassette transporter (ABCG1) on the transcription activity,and the relationship of the polymorphisms with the susceptibility to coronary artery disease (CAD).Methods A case-control study was conducted,217 CADpatients and 142 controls were enrolled in this study.Thesingle nucleotide polymorphisms (SNPs) in the promoter of ABCG1 were identified by sequencing.The promoter haplotypes of ABCG1 were determined with allele specific primer sequencing or Gene cloning sequencing.The transcription activity of the promoter haplotypes were evaluated with dual luciferase reporter system.The frequency of SNPs and haplotypes were analyzed between CAD group and the control group,premature CAD and non-premature CAD group,as well as multivessel lesion and single vessel lesion group.The frequency distribution was compared between two groups with x2 test or Fisher exact test.The difference of the luciferase activity was compared between groups by t-test or one-way analysis of variance.Results Only 3 SNPs were found in ABCG1 promoter sequence of about 1 000 bp upstream of the transcription start site,which are-384 (A/G),-204 (A/C) and-134 (T/G),respectively.The 3 SNPs are in strong linkage disequilibrium,Tajima's D =2.655 (P ＜ 0.01),which constituted 3 haplotypes.There was no significant difference in SNPs and haplotype frequency between the CAD group and the normal control group,and the severity of vascular disease and the early onset of coronary heart disease were not associated with the polymorphisms in ABCG1 promoter.There was no significant difference in the transcriptional activity of the three constitutive promoter haplotypes,but the transcriptional activity was notably elevated as the GAT haplotype was mutated into GAG (P ＜ 0.05).Conclusions The 3 SNPs identified in ABCG1 promoter region A did not alter the promoter activity.There was no significant correlation between the frequency distribution of SNPs and promoter haplotypes and the susceptibility to CAD.

Objective To establish a HPLC method for determination of naringenin and apigenin in Premna fulva. Methods The SHISEIDO￣SPOLAR C18(250 mm×4.6 mm,5μm) was used as analytical column.The mobile phase consisted of methanol￣0.2% phosphoric acid (42:58) with isocratic elution at a flow rate of 1.0 mL.min-1 .The detection wavelength of naringenin and apigenin was 288 nm and 340 nm, respectively.Column temperature was set at 35 ℃ , the injection Volume was 10 μL. Results Naringenin and apigenin had a good linear relationship in the concentration range of 0.180 ~ 3.60 μg (r =0.999 9) and 0. 0052 ~ 0. 1040 μg ( r = 0. 999 9), respectively. Conclusion The method is accurate and reliable. It is appropriate for the quantitative determination of naringenin and apigenin in Premna fulva and its preparations.