The aim of this study was to look for the chemical constituents of the bulbs of Fritillaria anhuiensis S.C.Chen et S.E.Yin. The bulbs of Fritillaria anhuiensis were extracted with 95% EtOH at reflux. Isolation and purification were performed by silica gel column chromatography. Structures of pure compounds were established on the basis of spectral analysis. Three compounds were obtained and identified as 12,15-epoxy-8(17),13-labdadien-19-ol (1), ent-3β-acetoxy-kauran-16β,17-diol (2), ent-kaurane-3β,16β,17-triol (3). Compound 1 is a new labdane-type diterpenoid. Compounds 2 and 3 were obtained from Fritillaria anhuiensis for the first time.

Cadaver preserved in formalin may develop pigmentation in the epidermis astime goes on.What is the nature of the colour substance?Are they degradationproduct of hemoglobin,melanin-like substance or others?In this paper,we reportthe results of observation on the pigmented substance from cadaver skin and charac-terize it by chemical and physical means.The epidermal layer of pigmented skin of cadaver was hydrolyzed by aceticacid,the hydrolysate was extracted with water and organic solvent in steps,thusthree portions were obtained:acidic water soluble,organic solvent soluble and inso-luble black precipitate.The later was identified as melanin by infrared spectrumanalysis,and chromatographic behavior after alkali fuse.The pigmented substancein acidic water and organic solvent are not identified yet.We also tried to find out the degradation products of hemoglobin,but wecouldn't detect them either by spectrum analysis or by crystallization of haem chlo-ride derivative.Hence,we intend that the deep colour of the cadaver skin after formalintreatment might be closely related to melanin,the mechanism of its formation isdiscussed.

Objective:To investigate the effect of repetitive transcranial magnetic stimulation (rTMS) on the hippocampal lipidome in a rat model of chronic unpredictable stress(CUS).Methods:Twenty-four SD rats were randomly assigned to the following 3 groups ( n=8 for each group): sham group, CUS group and CUS+ rTMS group. The sham group received only sham stimulation and rats in the CUS and CUS+ rTMS group were subjected to CUS stimulation. Then, rats received 5 Hz rTMS (5 Hz, 1.26 Tesla) or sham rTMS for 7 days. After the last stimulation, all rats underwent sucrose preference test, open filed test and forced swimming test so as to observe the effect of rTMS on depressive behavior. Then, rats were sacrificed, and the levels of lipid composition in hippocampus were determined by high performance liquid chromatography mass spectrometry and analyzed by lipid search software version 4.1 and SIMCA-P 14.1.The software of SPSS 19.0 was used for statistical analysis. Univariate analysis of variance was used for comparison among groups, and Tukey test was used for multiple comparison. Results:(1)There were significant differences in open field test, sugar preference test and forced swimming test among the three groups( F=6.853-7.466, all P<0.05). In the open field experiment, the exploring time and percentage of movement distance in central area of rats in CUS group((50.72±6.38)s, (11.41±1.55)%) was significantly less than that of sham group ((86.06±7.31)s, (18.60±1.21)%) and CUS+ rTMS group((79.87±7.87)s, (16.74±1.27)%)(all P<0.05). The results of sucrose preference test showed that the percentage of sucrose intake of rats in CUS group ((37.63±6.06)%) was significantly lower than that in sham group ((68.30±6.39)%) and CUS+ rTMS group ((62.68±5.50)%)(both P<0.05) . In forced swimming test, the immobility time of rats in CUS group ((137.60±13.36)s) was significantly longer than that of sham group ((80.57±10.36)s)) and CUS+ rTMS group ((86.14±11.49)s) (both P<0.05). (2)The levels of lipid composition in hippocampus were significantly different in the three groups( F=3.826-15.440, all P<0.05). The contents of phosphatidylethanolamine (PE) ((20 850±956.56)×10 7, (24 133.33±1 242.04)×10 7), phosphatidylinositol (PI) ((788.78±136.11)×10 7, (953.65±131.26)×10 7), lysophosphatidylcholine (LPC) ((340.29±35.66)×10 7, (275.32±35.78)×10 7), creatine phosphate (CerP) ((239.65±18.14)×10 7, (293.82±38.28)×10 7), sphingosine (So) ((22.96±4.04)×10 7, (15.36±3.87)×10 7), diglyceride (DG) ((3.35±0.85)×10 7, (4.57±1.02)×10 7) and monoglyceride (MG) ((6.71±0.82)×10 7, (7.94±0.91)×10 7)in hippocampus of rats in CUS group were significantly higher than those of sham group(all P<0.05), while the phosphatidic acid(PA) ((424.52±33.38)×10 7, (509.22±42.09)×10 7) and acyl carnitine(AcCa) ((2.68±0.33)×10 7, (3.39±0.33)×10 7) decreased(both P<0.05). Compared with CUS group, the contents of PE(21 816.67±928.26)×10 7, PI(83.16±91.52)×10 7, LPC(323.59±33.91)×10 7, CerP(236.39±32.02)×10 7, So(23.35±4.46)×10 7, DG(3.16±0.85)×10 7 and MG(7.03±0.26)×10 7 in the hippocampus of CuS+ rTMS group decreased, while the contents of PA(421.55±44.28)×10 7 and ACCA(2.56±0.32)×10 7 in the hippocampus of CUS+ rTMS group increased (all P<0.05). Conclusion:The levels of glycerophospholipids, glyceroglycerides, sphingolipids, fatty acids and other lipids in the hippocampus of CUS model rats are abnormal. And the 5 Hz rTMS intervention can ameliorate the depression like behavior and the disturbances of lipid in hippocampus of CUS model rats.

Objective:To investigate the effect of early intervention with electroacupuncture (EA) on the gut microbiota in a mouse model of post-traumatic stress disorder(PTSD).Methods:Totally 32 C57BL/6 mice were randomly assigned to the following 4 groups ( n=8 for each group): Control group, EA group, PTSD group and PTSD+ EA group.After 7 days acclimation, mice in the PTSD group and PTSD+ EA group were subjected to modified single prolonged stress (mSPS). Mice in the EA group and PTSD+ EA group received EA (2/15 Hz, 1 mA, dilatational wave, 30 min/d) on "Baihui" for 7 days. Mice in the Control group and PTSD group received false stimulation (stimulated the same acupiont without electricity) for 7 days. Seven days after the last stimulation, elevated plus maze test and fear conditioning test were conducted to observe the effect of EA on PTSD-like behavior of mice. At the same time, feces of the mice were collected for gut microbiota detection by 16S rRNA sequencing.SPSS 19.0 was used for statistical analysis.One-way ANOVA was used for multiple group comparison and Bonferrani test was done for further pairwise comparision. Results:(1) There were statistically differences in the open arm activity time of the elevated plus maze test and the immobility time in contextual and cued fear conditioning test among the four groups ( F=6.93, 5.26, 14.51, all P<0.01). In the elevated plus maze test, mice in PTSD group ((60.17±15.52) s) showed significant less time in the open arms than mice in Control group((96.37±14.62) s) and PTSD+ EA group ((86.89±15.02) s) (both P<0.05). In the fear conditioning test, mice in PTSD group ((121.99±29.67) s, (130.82±29.11) s) showed significant increased immobility time both in contextual and cued fear conditioning tests than mice in Control group((74.50±26.65) s, (39.50±23.52) s) and PTSD+ EA group ((76.77±22.60) s, (102.17±3.39) s)(both P<0.05). (2) There were no significant differences among the four groups in the alpha diversity of gut microbiota ( F=0.79-2.45, all P>0.05). (3)Correlation analysis showed that 13 gut microbiotas were negatively correlated with the immobility time in contextual fear conditioning test, 2 gut microbiotas were positively correlated with it; 7 gut microbiotas were negatively correlated with the immobility time in cued fear conditioning test, 1 gut microbiota was positively correlated with it; 3 gut microbiotas were positively correlated with time spent in open arms of elevated plus maze test. Conclusion:Early intervention with EA can improve anxiety-fear like behaviors and gut microflora disorder in PTSD model mice.

Objective To construct mutant recombinant vector of epidermal growth factor receptor ( EGFR) gene G719S and T790M sites associated with cervical cancer, lay the foundation for the detection of EGFR gene mutation in cervical cancer. And using it to es-tablish a molecular switch platform to detect cervical cancer EGFR gene mutations. Methods Using the wild-type recombinant plasmid as template, the mutant fusion target fragment were amplified by overlap PCR, then connect this target fragment into the vector pMD19-T. The constructed mutant recombinant plasmid was finally transformed into competent cells E.coli DH5αfurther identified by PCR with bacterial solution and genome sequencing. Establishing the molecular switch for the detection of clinical cervical cancer samples. Re-sults The G719S and T790M mutations were successfully certified by genome sequencing, and the site-directed mutant vector was successfully constructed. In addition, a molecular switch detection platform was also successfully established for the detection of cervical cancer tissue DNA. Conclusion We successfully constructed an EGFR gene mutant recombinant vector by overlap PCR technique, which providing a new technical means for gene site-directed mutagenesis. And the molecular switch detection platform was successfully established based on it, which furnishing a new method for clinical detection of EGFR gene mutations.

Objective:To construct a nomogram model that can be used to predict thrombocytosis after splenectomy in patients with traumatic splenic rupture.Methods:The clinical data of 54 patients with splenectomy in Lu′an People′s Hospital from November 2016 to November 2021 were retrospectively analyzed. Among them, postoperative thrombocytosis occurred in 33 cases (postoperative thrombocytosis group), and 21 cases had no significant changes in platelets (postoperative platelet normal group). The general clinical data were recorded; the platelet parameters 14 d after operation were measured, including platelet count, mean platelet volume (MPV), procalcitonin (PCT) and platelet distribution width (PDW). The receiver operating characteristic (ROC) curve was drawn to analyze the predictive value of platelet parameters for thrombocytosis after splenectomy in patients with traumatic splenic rupture. Multivariate Logistic regression analysis was used to analyze independent risk factors of thrombocytosis after splenectomy in patients with traumatic splenic rupture. A nomogram model to predict thrombocytosis after splenectomy in patients with traumatic splenic rupture was established with R language software 4.0.2 package, internal validation of the nomogram model was performed using correction curves, and the prediction efficiency of the nomogram model was evaluated using decision curves.Results:The incidence of shock, rate of blood transfusion volume≥2 000 ml, platelet count and PCT in postoperative thrombocytosis group were significantly higher than those in postoperative platelet normal group: 69.70% (23/33) vs. 19.05% (4/21), 66.67% (22/33) vs. 38.10% (8/21), (823.56 ± 129.81) ×10 9/L vs. (521.92 ± 85.89) ×10 9/L, (0.87 ± 0.11)% vs. (0.54 ± 0.09)%, the MPV and PDW were significantly lower than those in postoperative platelet normal group: (10.23 ± 1.03) fl vs. (11.57 ± 0.92) fl and 0.113 ± 0.012 vs. 0.125 ± 0.020, and there were statistical differences ( P<0.01 or <0.05). ROC curve analysis results show that the area under curve of platelet count, MPV, PCT and PDW in for predicting the thrombocytosis after splenectomy in patients with traumatic splenic rupture were 0.973, 0.835, 0.987 and 0.734, and the optimal cut-off values were 642.29 ×10 9/L, 11.02 fl, 0.7% and 0.120. Multivariate Logistic regression analysis result showed that the platelet count, MPV, PCT, PDW, shock and blood transfusion volume were independent risk factors for thrombocytosis after splenectomy in patients with traumatic splenic rupture ( OR = 1.571, 1.243, 1.042, 1.413, 1.436 and 1.726; 95% CI 0.014 to 1.762, 0.743 to 2.862, 0.954 to 2.563, 0.584 to 2.389, 0.045 to 2.643 and 0.154 to 2.143; P<0.01 or <0.05). When platelet count, MPV, PCT, PDW, shock and blood transfusion volume were included as predictors for constructing the nomogram model, the internal validation results showed that the C-index of the nomogram model for predicting thrombocytosis after splenectomy in patients with traumatic splenic rupture was 0.793 (95% CI 0.267 to 2.311); the threshold value of the nomogram model for predicting thrombocytosis after splenectomy in patients with traumatic splenic rupture was >0.067, and the nomogram model provided a net clinical benefit; the clinical net benefit of the nomogram model was greater than that of platelet count, MPV, PCT, PDW, shock and blood transfusion volume. Conclusions:The nomogram model based on platelet count, MPV, PCT, PDW, shock and blood transfusion volume that affect the high risk of thrombocytosis after splenectomy in patients with traumatic splenic rupture has great clinical value in screening and identifying high risk patients.

Objective: To explore the association between the single nucleotide polymorphism (SNP) of rs175048 in ataxia telangiectasia mutated (ATM) gene and lung cancer susceptibility in Han population. Methods: A total of 225 cases of blood samples from lung cancer patients treated in Hospital of Traditional Chinese Medicine of Hengyang City and the Affiliated First Hospital of Nanhua University from October 2015 to August 2016 were collected as case group, and 128 cases of blood samples from healthy people were collected as the control. The polymorphisms of ATM rs175048 of above mentioned participants were detected by using the SNP sensitive On/Off Switch technique. The genotypes and allele frequencies were analyzed to compare the distribution difference between case group and control group as well as its association to the clinical features of lung cancer. Results: The genotype frequencies of AA, AT and TT of ATM rs175048 were 24.9%, 52.9%, 22.2% in case group and 42.2%, 42.2%, 15.6% in control group, respectively (all P< 0.01). Moreover, the frequencies of alleles A and T were 51.0%, 49.0% in case group, and 63.0%, 37.0% in control group (all P<0.01). Genotype TT might increase while genotype AT might decrease the risk of lung cancer. rs175048 SNP was significantly correlated with smoking, age, sex and family history (all P<0.05). Conclusion: rs175048 SNPis significantly associated with lung cancer, and TT genotype may increase the risk of lung cancer.