This study was designed to investigate the potential protective effect of isopimpinelline against para-chlorophenylalanine(PCPA)-induced pineal gland damage in rats.Forty male Sprague-Dawley(SD)rats were divided into four groups(n=10 each):a normal group,a model group,a melatonin-treated group(10 mg/kg),and an isopimpinelline-treated group(1.5 mg/kg).All groups,except for the normal,received intraperitoneal injection of PCPA(450 mg/kg)to induce pineal gland damage.Subsequent treatments were administered orally for 7 days.Sleep latency and duration were evaluated on the sixth day using the pentobarbital sodium sleep synergy test.After the treatment period,serum melatonin levels and pineal gland inflammation markers were assessed alongside oxidative and antioxidative parameters.Histological examinations of the pineal gland were conducted,and the expression of proteins related to the Nrf2 and NF-κB signaling pathways were quantified.Data showed that isopimpinelline alleviates the structural damage in the pineal gland of model rats,significantly elevated serum melatonin levels,and markedly improved sleep latency and duration(P＜0.05).Isopimpinelline activated the Nrf2 signaling pathway by inhibiting Keap1 expression,which facilitated the nuclear translocation of Nrf2 and upregulated the antioxidant proteins NQO1 and HO-1,thereby mitigating oxidative stress in the pineal gland(P＜0.05).Furthermore,isopimpinelline significantly reduced the levels of pro-inflammatory cytokines IL-2,TNF-α and IL-6.Isopimpinelline also suppressed the NF-κB signaling pathway,reducing the expression of NF-κB p65,IKKβ,and p-IKKβ proteins,as well as the nuclear translocation of NF-κB p65(P＜0.05),thereby providing anti-inflammatory benefits.In conclusion,isopimpinelline could protect pineal gland from damage by activating the Nrf2 signaling pathway and inhibiting the NF-κB pathway.

【Objective】 To establish and verify the vacuum decay method for the tightness inspection of blood products. 【Methods】 The method for inspecting the tightness of blood product was established, and the detection limit, linearity, range, accuracy, precision and durability were verified according to the requirements of methodological verification.The validated method was used to check the tightness of blood product packaging. 【Results】 The detection limit of this method was 2.5 μm, linear correlation coefficient was r=1, the differential pressure of positive sample was within the allowable range of accuracy, and the durability met the requirements.The RSD of results of 6 repeatability tests and 12 intermediate precision tests were both less than 10%, and all validation items met the verification standards. 【Conclusion】 Vacuum decay method can be used to check the tightness of blood products.

【Objective】 To establish gas chromatography for the determination of tributyl phosphate(TBP) residues in human prothrombin complex and then verify it. 【Methods】 Acid modified polyethylene glycol(PEG)(20M) capillary column was used with n-hexane as solvent. The chromatographic parameters were as follows: gasification chamber temperature at 220 ℃, column temperature at 155 ℃, detector temperature at 220 ℃, column flow rate at 2.0 mL/min, carrier gas as N2, detector as FID, and collection time for 10min. The accuracy, repeatability, linearity, specificity, intermediate precision, detection limit, quantitative limit, range and durability were verified. 【Results】 The verification results showed that the method had good specificity. The linear regression correlation coefficient of standard curve was 0.999 90. The recovery rate were 98.4%, 97.5% and 95.7% when the concentration at 50%, 100%(30μg/mL) and 150%, respectively, with an average recovery of 97.2% and a relative deviation of 2.15%. When the concentration was 100%, the repeatability was 2.08%, and the relative deviation of intermediate precision was 1.63%. The detection limit was 0.255 μg/mL, and the quantitative limit was 0.511 μg/mL. After changing capillary chromatographic columns with different batch numbers but the same types and manufacturer, the applicability test of the system met the requirements, and the method had good durability. 【Conclusion】 This method can be used for the determination of TBP residues of human prothrombin complex in laboratory.

OBJECTIVE：To study the improvement effects of isopimpinelline on p-chlorophenylalanine（PCPA）-induced pineal injury model rats and its effect on expression of biological clock gene. METHODS ：Totally 60 rats were divided into blank control group（2％ polysorbate solution），model control group （2％ polysorbate solution），positive control group （melatonin，10 mg/kg） and isopimpinelline high-dose ，medium-dose and low-dose groups （3，1.5，0.75 mg/kg）. Except for blank control group ，rats in other groups were given PCPA intraperitoneally （450 mg/kg）to establish pineal injury model. After modeling finished ，they were given relevant medicine intragastrically ，once a day ，for consecutive 7 d. On the 6th day of administration ，the sleep latency and sleep duration of rats in each group were investigated by pentobarbital sodium coordination sleep test ；after last administration ， ELISA assay was used to determine the serum level of melatonin in rats. Fluorescence microscope and electron microscope were used to observe the pathological tissue and cell ultrastructure changes of the pineal gland. RT-qPCR was used to detect the mRNA expressions of biological clock gene Clock，Bmal1，Per1，Per2，Per3，Cry1，Cry2 in pineal gland of rats. RESULTS ：Compared with blank control group ，model control group had significantly longer sleep latency （P＜0.05）；serum melatonin ，mRNA expressions of Bmal1 and Per1 in pineal gland were significantly decreased （P＜0.05 or P＜0.01）while mRNA expression of Per3 was increased significantly （P＜0.05）. The pineal gland cell arrangement disorder ，nuclear pyknosis ，vacuolar degeneration increased and cell number decreased significantly ；mitochondria swollen ，cristae broken and pyknosis were observed. Compared with model control group ，the sleep latency of isopimpinelline high-dose group was shortened significantly （P＜0.05），sleep duration time was prolonged significantly （P＜0.05）；the levels of melatonin in serum ，mRNA expressions of Clock，Bmal1， Per1，Cry1 and Cry2 in pineal gland of rats were increased significantly （P＜0.05 or P＜0.01）. In isopimpinelline medium-dose group，the sleep latency was shortened significantly （P＜0.05）；the levels of melatonin in serum and mRNA expressions of Clock， Bmal1，Per1，Cry1，Cry2 in pineal gland were increased significantly （P＜0.05 or P＜0.01），while mRNA expression of Per3 was decreased significantly （P＜0.05）. In isopimpinelline low-dose group ，the levels of mRNA expressions of Clock，Bmal1，Per2 and Cry2 were increased significantly （P＜0.05），while mRNA expression of Per3 was decreased significantly （P＜0.05）. Cell arrangement disorder was improved and nuclear pyknosis vacuole degeneration was decreased to some extent in isopimpinelline groups；mitochondria swelled ，cristae fractured ，and pyknosis decreased to some extent. CONCLUSIONS ：Isopimpinelline can improve PCPA-induced pineal gland injury in rats ；it can up-regulate the expressions of positive regulators Clock，Bmal1 and negative regulators Per1，Per2，Cry1，Cry2，while down-regulate the expression of negative regulator Per3.

OBJECTIVETo evaluate the clinical effects on incomplete intestinal obstruction treated with the adjuvant therapy of acupuncture and moxibustion.

METHODSUsing the retrospective analysis, 80 patients of incomplete intestinal obstruction were divided into an observation group and a control group, 40 cases in each one. In the control group, the routine treatment was given, such as fasting, gastrointestinal decompression, parenteral nutrition, infection prevention with antibiotics and enema laxative. In the observation group, on the basis of the treatment as the control group, acupuncture was applied at bilateral Zusanli (ST 36), Shangjuxu (ST 37) and Xiajuxu (ST 39); moxibustion was used at left Yangchi (TE 4), Zhongwan (CV 12), Qihai (CV 6) and Guanyuan (CV 4). The treatment was given once a day, 30 min each time. The average days of treatment, the surgical transfer rate, the time to first flatus, the recovery time of defecation and the time of solid food intake were observed in the patients of the two groups.

RESULTSThe average days of treatment in the observation group was obviously less than that in the control group (<0.05). The surgical transfer rate in the observation group was obviously lower than that in the control group (<0.05). The time to first flatus, the recovery time of defecation and the time of solid food intake were all obviously earlier than those in the control group (all <0.05).

CONCLUSIONThe adjuvant therapy of acupuncture and moxibustion achieves the significant therapeutic effects on incomplete intestinal obstruction, shortens the treatment duration and reduces the surgical transfer rate and the patient's economic burden.

Objective To investigate the effect of miR-132-3p on the proliferation of endothelial progenitor cells and its regulatory mechanism in order to provide a new theoretical basis for the treatment of deep venous thrombosis. Methods Real-time quantitative PCR (qPCR) was used to detect the expression of miR-132-3p in the plasma and endothelial progenitor cells of 27 healthy volunteers and 22 thrombus patients, and in endothelial progenitor cells under normoxic and hypoxic conditions. The miR-132-3p analogue and the specific siRNA were transferred into endothelial progenitor cells by the electroporation method. The effect of miR-132-3p on the proliferation of endothelial progenitor cells was detected using MMT and Cell Counting Kit-8 (CCK-8) methods. The effects of miR-132-3p on the expression of FOXO1 in endothelial cells were analyzed using the luciferase assay and western blots. Results The expression of miR-132-3p in clinical patients with thrombosis was significantly decreased to 0.45 ± 0.05 times of that of the healthy volunteers (P＜0.05). The expression of miR-132-3p in endothelial progenitor cells under hypoxia was down-regulated to (0.23 ± 0.13) times of that of under normoxia (P＜0.05). The expression of miR-132-3p of experiment group under hypoxia was up-regulated to (15.72 ± 2.06) times of that of control group (P＜0.05). MMT assay showed that the proliferation of cells in the experimental group under hypoxic condition was up-regulated to (7.79 ± 1.37) times of that in the control group (P＜0.01). CCK-8 assay showed that cell proliferation in experimental group was up-regulated to (6.46 ± 0.38) times of that in the control group (P＜0.01). Software analysis showed that FOXO1 was a direct target of miR-132-3p. Luciferase activity of miR-132-3p mimics transfected endothelial progenitor cells under hypoxic conditions were 0.47 times of that in siRNA treatment group. Western blot showed that the expression of FOXO1 protein in endothelial progenitor cells transfected with miR-132-3p mimics in hypoxia was 0.18 times of that in siRNA treatment group. Conclusions Compared with healthy volunteers, miR-132-3p expression in the blood of patients with thrombosis was significantly reduced that can promote transcription of the FOXO1 gene (and protein expression) and inhibit the proliferation of endothelial progenitor cells. It could be closely related to the formation of venous thrombosis.

Objective To investigate the psychological status of diarrhea type irritable bowel syndrome (D-IBS) patients overlapped with functional dyspepsia (FD) and its relationship with the changes of brain-gut peptides and cytokines levels.Methods A total of 190 D-IBS FD overlapped patients and 30 healthy controls were selected and all of them completed the questionnaire.The adult psychological testing system was applied for the Symptom Checklist (SCL-90) assessment.Plasma levels of 5-hydroxytryptamine (5-HT),somatostatin (SS),vasoactive intestinal peptide (VIP),endothelin (ET),interleukin (IL)-10 and IL-12 were measured in 59 randomly selected patients.Single factor analysis of variance and LSD test were performed for statistical analysis.Results In D-IBS-FD overlapped patients,the plasma 5 HT levels of the D-IBS patients and the FD patients were (1906.83±676.88) μg/L and (1745.53±653.23) μg/L respectively,both of them were significant higher than those of control group [(1289.44±918.45) μg/L,LSD test,both P＜0.05].However the levels of VIP and IL-10 were significant lower than those of control group (LSD test,both P＜0.01).In D-IBS patients,the plasma level of SS was significantly lower than that of control group (LSD test,P=0.025); the level of ET was significantly higher than that of control group and FD patients (LSD test,both P＜0.05).In D-IBS-FD overlapped patients,the scores of anxiety,depression and opponent were higher than those of control group (F=5.114,4.555 and 10.285,all P＜0.05).The plasma VIP and SS levels of patients with anxiety or depression were lower than those of control group (LSD test,both P＜0.05).The plasma VIP level was lower than that of patients without psychological disorders (LSD test,P=0.005).The plasma ET level of patients with disease course over five years was significantly higher than that of patients with disease course less than one year or between one and five years (LSD test,both P＜0.05),and the IL-10 level was significantly lower than that of patients with disease course less than one year (LSD test,P=0.004).Conclusions There were sub-healthlated with disease course,psychological disorder,the increased plasma 5-HT level and the decreased IL-10 level.There was close correlation between the lower gastrointestinal symptoms and the decreased levels of VIP,SS and the increased level of ET.

Objective To compare the clinical features and severity of thoracoabdominal combined injuries(TACI) and thoracoabdominal multiple injuries(TAMI) so as to guide the diagnosis and management of trauma. Methods A total of 167 cases of chest trauma complicated by abdominal trauma were reviewed and divided into two groups. Group TACI(132 cases) was associated with diaphragmatic rupture but group TAMI(35 eascs)not. The injury features and the clinical characteristics of both groups were analyzed and the trauma severity of two groups e-valuated by trauma score system. Results The overall mortality was 9.6 %(16/167)of all the cases. The mortality was 14.3% in group TACI (5/35)and 8.3% in group TAMI(11/132). There was no significant difference in revised trauma score(RTS) and thoracic Abbreviated Injury Scale(MS) between the two greups(P> 0.05),but the patients in the TACI group had higher Glasgow Coma Scale(C, CS)and abdominal AIS (P < 0.05), and the patients in the TACI group had higher injury severity scale (ISS) (P < 0.01). Conclusion TACI and TAMI have differ-ence in clinical characteristics,injury severity and treatment results. So it is necessary to take different measurements to reduce the complica-tions and improve the prognosis.

For the purpose of diagnosis of sudden manhood death syndrome, immu- nohistochemical study of ANP was performed in right atria of 10 cases of sudden manhood death syndrome (SMDS) and 10 cases of noncardiac death controls with LSAB-method. It was found that the ANP granulus in right atria of SMDS were obviously depleted, compared with that in the control groups. The results showed that depletion of ANP granules in atria plays an important role in the causes of death of SMDS. This experiment also provides a new approach for studying the causes of SMDS.

Objective: To observe the soluble and dispersive characteristics of Liuwei Dihuang Ultra micro power. Methods: The similarity and difference between the soluble and dispersive characteristics of two powders with HPLC atlas were observed. The contents and dissolving characteristic of target compositions Ursolic acid and Paeonol of Liuwei Dihuang powder and ultra micro powder were compared and studied. Results: At the same retaining time two powders had the same absorbent peak, but the peak values of two powders had apparent difference. the ursonic acid and paeonol in ultra micro powder were 44.55% and 7.6% higher than those of the normal powder respectively. Conclusion: After Liuwei Dihuang is broken into ultra micro power, the chemical compositions do not change but the dissolving speed and contents increase greatly.