ObjectiveTo explore the application value of modified Wendantang in the treatment of perimenopausal insomnia of phlegm heat type and observe its effects on neurotransmitters and body fat. MethodsA total of 162 patients with perimenopausal insomnia of phlegm heat type who were treated in the Traditional Chinese Medicine Department of The First Affiliated Hospital with Nanjing Medical University from May 2021 to November 2023 were randomly divided into two groups before inclusion in the study. The control group of 81 cases received conventional Western medicine treatment， while the observation group of 81 cases received combined treatment with modified Wendantang， and the treatment lasted for 4 weeks. ResultsInterleukin-β （IL-1β）， and tumor necrosis factor alpha （TNF-α） decreased， while serum dopamine （DA）， melatonin （MT） and serotonin （5-HT） increased in both groups after treatment compared with before treatment （P<0.05）. After treatment， compared with the control group， the observation group showed better improvement in the above indicators （P<0.05）. The levels of follicle stimulating hormone （FSH） and luteinizing hormone （LH） decreased， while the level of estradiol （E₂） increased in both groups after treatment compared with before treatment （P<0.05）. Compared with the control group， the observation group showed better improvement in the above sex hormone indicators after treatment （P<0.05）. The rapid eye movement sleep period （REM） and total sleep time （TST） were prolonged， and the sleep latency （SL） was shortened in the two groups after treatment compared with before treatment （P<0.05）. Compared with the control group， the observation group showed better improvement in sleep conditions after treatment （P<0.05）. The Pittsburgh sleep quality index （PSQI） and Traditional Chinese Medicine （TCM） total symptom score decreased significantly in both groups after treatment compared with before treatment （P<0.05）. Compared with the control group， the observation group showed better improvement in PSQI and TCM total symptom score after treatment （P<0.05）. The total effective rate of the observation group was 93.83% （76/81）， while that of the control group was 82.72% （67/81）， and the two had a statistically significant difference （χ²=9.135， P<0.05）. ConclusionFor patients with perimenopausal insomnia of phlegm heat type， the application of modified Wendantang can help regulate neurotransmitters and inflammatory factor levels， improve sex hormone levels， enhance sleep quality， and improve therapeutic effects.

Trihelix transcription factors play important roles in plant light responses, growth and development, and stress responses. However, Trihelix has not yet been reported in Eucommia ulmoides. In this study, bioinformatics methods were used to comprehensively identify and analyze the expression patterns of the Trihelix gene family in E. ulmoides, aiming to provide a basis for further functional studies of EuGTs genes. A total of 9 Trihelix gene family members were identified in E. ulmoides, encoding proteins with 339 to 883 amino acids, with isoelectric points ranging from 5.13 to 9.39 and relative molecular weights between 36 992.06 and 97 871.61. Subcellular localization results showed that only EuGT-2 was localized in chloroplasts, while the others were located in the nucleus. The Trihelix gene family was categorized into six subfamilies: GT-1, GT-2, SH4, SIP1, GTγ, and GTδ. EuGTs were distributed among three subfamilies: SH4, GT-1, and GT-2, containing 1, 6, and 2 Trihelix proteins, respectively, with 2 to 17 exons. The promoters of EuGTs contained various cis-acting elements related to hormones, stress, photoperiod, and growth and development. Collinearity analysis revealed 5 collinear gene pairs between E. ulmoides and Arabidopsis thaliana, and 14 collinear gene pairs between E. ulmoides and Populus. Expression pattern analysis showed that EuGTs exhibited tissue-specific expression: EuGT-1, EuGT-2 had the highest expression levels in leaves, EuGT-4, EuGT-6, EuGT-9 had the highest transcriptional levels in marginal peel, and EuGT-5、EuGT-8 were predominantly expressed in the xylem. As leaves developed, EuGTs showed a trend of asynchronous changes. No significant differences in EuGTs expression were observed between male and female flowers, with high expression levels mainly during the induction stage of flowering. The qRT-PCR analysis indicated that most EuGTs genes were most highly expressed in the leaves of E. ulmoides, while EuGT-5 was highly expressed in the stems. Under 200 mmol·L~(-1) NaCl treatment, most EuGTs genes exhibited an initial increase followed by a decrease in expression, significantly responding to salt stress. This study provides important genetic resources for further exploration of EuGTs gene functions and germplasm innovation in E. ulmoides.
Plant Proteins/metabolism* ; Gene Expression Regulation, Plant ; Eucommiaceae/chemistry* ; Phylogeny ; Multigene Family/genetics* ; Gene Expression Profiling ; Transcription Factors/metabolism* ; Genome, Plant/genetics*

OBJECTIVE to explore the material basis and potential molecular mechanism of Ziziphi Spinosae Semen- Acori Tatarinowii Rhizoma in the treatment of insomnia using network pharmacology and molecular docking, and establish insomnia mouse model by p-chlorophenylalanine(PCPA) for verification in vivo.METHODS Firstly, the chemical constituents of Ziziphi Spinosae Semen and Acori Tatarinowii Rhizoma were collected through TCMSP database and the potential active constituents were screened. The genes related of insomnia were obtained from GeneCards, OMIM and TTD databases, and the intersection targets of Ziziphi Spinosae Semen-Acori Tatarinowii Rhizoma and insomnia diseases were obtained. Then the network map of Chinese medicine-compound-target-disease was constructed in Cytoscape 3.6.1 software. The protein interaction network diagram was established in the STRING database. Functional enrichment analysis of target GO and KEGG pathways were performed using the DAVID database. Then the molecular docking technology was used for preliminary verification. The 60 male ICR mice were randomly divided into normal group, model group, high, medium and low dose groups(8.0, 4.0, 2.0 g·kg-1) and diazepam group(3 mg·kg-1). In addition to the normal group, PCPA(30 mg·mL-1) was intraperitoneally injected on the first and second days to establish the insomnia model. Then the drug was administered continuously for 7 d, and the normal group and the model group were given the same volume of normal saline. The sleep latency and duration of mice induced by the upper threshold dose of pentobarbital sodium(55 mg·kg-1), vertical and horizontal scores in the behavioral open-field experiment, open arm entry(OE%) and open arm time(OT%) of elevated cross maze were determined. HE staining was used to observe the hypothalamic histopathological situation. Serum levels of TNF-α and CASP3 were detected by ELISA. Finally, Western blotting was used to detect the expression of AKT1, p-AKT1 protein in the hypothalamus of the mice in each group. RESULTS The potential active components of Ziziphi Spinosae Semen and Acori Tatarinowii Rhizoma were 9 and 5, and the common targets with insomnia were 34. A total of 160 GO items were obtained through GO enrichment analysis. KEGG pathway analysis found that the signaling pathway was mainly related to inflammatory signaling pathway, among which AKT1, CASP3 and TNF were the key targets. The results of molecular docking showed that the selected compounds had high binding activity with the key targets. Animal experiment results showed that the Ziziphi Spinosae Semen-Acori Tatarinowii Rhizoma for insomnia could significantly shorten the model mice sleep latency, prolong sleep duration, reduce the vertical and horizontal score, improve the OE% and OT%, restore the hypothalamus pathological tissue damage, significantly reduce the content of TNF-α and CASP3, raise the level of AKT1 protein expression in the tissue of the hypothalamus. CONCLUSION Ziziphi Spinosae Semen-Acori Tatarinowii Rhizoma can regulate TNF signaling pathway by acting on TNF-α, CASP3, AKT1, p-AKT1 and other targets to treat insomnia.

Objective:To investigate the related risk factors related to patellar dislocation combined with osteochondral fracture (OCF).Methods:A case control study was conducted to analyze the clinical data of 278 patients with patellar dislocation admitted to Second Hospital of Lanzhou University from January 2016 to June 2020, including 98 males and 180 females; aged 12-41 years [(21.1±4.6)years]. The patients were divided into OCF group ( n=112) and non-OCF group ( n=166) according to the surgical records and occurrence of OCF or not under intraoperative arthroscopy. The data of the two groups were recorded, including basic characteristics (ie, sex, age, sides of injury, whether or not the epiphysis is closed), patella height, patellar inclination angle, tibial tubercle-trochlear groove (TT-TG), femoral trochlear shape, tibial lateral rotation angle and femoral anteversion angle. Univariate analysis was adopted to evaluate the correlation of the data with patellar dislocation combined with OCF. Multivariate Logistic regression analysis was used to determine the independent risk factors of patellar dislocation combined with OCF. Results:Univariate analysis showed that patellar dislocation combined with OCF was related to male, patellar inclination angle and femoral anteversion angle ( P<0.05 or 0.01), while not to age, epiphysis is closed or not, sides of injury, patella height, TT-TG, femoral trochlear shape and tibial lateral rotation angle (all P>0.05). Multivariate Logistic regression analysis showed that male ( OR=1.75, 95% CI 1.07-2.90, P<0.05) and increased femoral anteversion angle ( OR=3.12, 95% CI 1.89-5.17, P<0.05) were significantly correlated with patellar dislocation combined with OCF. Conclusion:Male and increased femoral anteversion angle are independent risk factors for patellar dislocation combined with OCF.

Respiratory virus remains to be an important pathogen of respiratory disease in children.The disease can occur in all age groups, especially in young children.Most viral infections have a good prognosis, but special viruses still cause great harm to the health of children.Respiratory viral therapy includes symptomatic therapy, broad-spectrum antiviral drugs, drugs that directly target the viral replication cycle, drugs that attenuate the inflammatory response, and anti-viral nanodrugs.New antiviral drugs are urgently needed to develop.The repurposing of the existing therapeutic agents previously designed for other virus infections is also an effective way.The treatment of respiratory virus infection has become an important topic in clinical research.

OBJECTIVE： To prepare standard decoction of fried Perilla frutescens seed and study the quality standard. METHODS： According to the preparation requirements of standard decoction， 17 batches of standard decoction of fried P. frutescens seed were prepared， and the yield of paste was calculated. HPLC method was used for quantitative analysis of rosmarinic acid in standard decoction of fried P. frutescens seed. The determination was performed on Agilent 5 TC-C18（2） column with mobile phase consisted of methanol-0.1％ formic acid solution （40 ∶ 60， V/V） at the flow rate of 1.0 mL/min. The detection wavelength was set at 330 nm， and column temperature was 30 ℃. The sample size was 5 μL. The transfer rate was calculated. HPLC fingerprint was established for 17 batches of standard decoction of fried P. frutescens seed， and analyzed by using TCM Chromatogram Fingerprint Similarity Evaluation System （2012 edition）. The chromatogram peaks were identified by comparing retention time of common peak. RESULTS： The yield of paste were 5.55％-9.75％ in 17 batches of standard decoction of fried P. frutescens seed. The contents of rosmarinic acid were 0.44％-1.58％， and average content was 1.08％. The transfer rates of rosmarinic acid were 18.31％-34.32％， and average transfer rate was 25.42％. In HPLC fingerprints for 17 batches of standard decoction of P. frutescens seed， a total of 11 common peaks were identified， and the similarity was higher than 0.95. Five common peaks were identified， namely caffeic acid （peak 3），luteolin （peak 5）， rosmarinic acid （peak 8） ， luteolin （peak 9）， apigenin （peak 10）. CONCLUSIONS： The quality control method of standard decoction of fried P. frutescens seed is established， which can provide reference for the formulation of the quality standard of fried P. frutescens seed granules and related preparations.

Objective To evaluate the efficacy of microsurgical techniques combined with bone transportation for osteomyelitis following open fracture on the leg.Methods From April 2007 to April 2016,25 patients with osteomyelitis caused by severe open injury on the leg were treated at Orthopaedic Department,General Hospital of Rocket Force.They were 22 men and 3 women,aged from 19 to 65 years (average,36.5 years).Their open injury was located at upper tibia in 6 cases,at middle tibia in 6 and at lower tibia in 13.Eight patients received internal fixation with plate and 17 unilateral external fixation.By the Cierny-Mader classification,their chronic osteomyelitis was of type Ⅲ in 14 cases and of type Ⅳ in 11.The tibial defects after excision of infectious segment ranged fromn 5 to 14 cm in length (average,8.8 cm).After infectious tissues were removed,antibiotic bone cement chains were inserted into the defects.The open wounds were covered with microsurgical pedicled or free flaps.Bone transportation with unilateral external fixation was used to deal with the bone defects after removal of the infectious bone.Results Bone infection was controlled in the 25 patients.The rate of infection control after one debridement procedure was 76.0％ (19/25),and that after 2 debridement procedures was 24.0％ (6/25).Flap repair led to early closure of the open wounds.The successful rate of wound closure after one operative procedure was 84.0％ (21/25),and that after secondary operation was 16.0％ (6/25).Bony union of tibial defects was achieved in all the 25 patients with no recurrence of osteomyelitis.The patients were followed up for 18 to 36 months (average,24.6 months).Re-fracture happened in one case and was treated with another operation.Bone union time starting from bone transportation ranged from 8 to 18 months (average,10.5 months).Time for external fixation ranged from 12 to 20 months (average,18.4 months).Conclusion For osteomyelitis caused by severe open injury on the leg,microsurgical techniques can be used to repair open wounds and bone transportation to reconstruct bone defects after removal of infectious bone.

Objective To report the clinical outcomes of latissimus dorsus flap used to reconstruct severe trauma on lower leg.Methods From May 2011 to September 2016,12 patients were treated for severe trauma on lower leg at Orthopaedic Department,General Hospital of Rocket Force.They were 10 men and 2 women,aged from 22 to 54 years (average,37.5 years).According to the Gustilo classification,their open fractures were of type ⅢA in 2 cases and of type ⅢB in 10.The areas of soft tissue defect ranged from 15 cm × 8 cm to 35 cm × 12 cm.Their wounds were covered with latissimus dorsus flaps;the dorsal expansion of the ankle joint was reconstructed after their bone infection was controlled.Two flaps were anastomosed directly to the anterior tibial vessels and one flap to the femoral vessels.The other 9 flaps were anatomosed to the lateral circumflex femoral artery,bridging with the greater saphenous vein (average length:32 cm).Ten flaps were neuroanatomosed with the motor branch of common peroneal nerve and 2 with the motor branch of vastus lateralis.The therapeutic outcomes were evaluated in terms of repair of soft tissue defects,control of infection and range of active dorsal flexion of the ankle at neutral position.Results The 12 patients were followed up for an average of 3.5 years (from 1 to 6 years).All the 12 flaps survived and infections were eradicated.The fractures were healed with muscle strength of grade 3 to grade 4.Their ankle extension was reconstructed.Their limbs were preserved with satisfactory limb function.The bone exposures of 5 cm × 3 cm and 6 cm × 3 cm in area after debridement respectively in 2 cases of Gustilo type ⅢA were covered with latissimus dorsus flaps.The range of active dorsal flexion of the ankle at neutral position increased from preoperative 10.5° to postoperative 19.1° on average.Conclusion In treatment of severe trauma on lower leg,latissimus dorsus flaps can be used to repair skin wound,control infection and reconstruct ankle function simultaneously,with advantages of reduced operative procedures,a high rate of limb preservation,and better function recovery.

Aim To study the analgesic effect of geraniol on neuropathic pain and to explore the possible mechanism.Method A neuropathic pain rat model of Spared Nerve Injury(SNI) was established to measure changes in the threshold of paw withdrawal before and after i.p.administration of geraniol.Patch clamp whole-cell recording was performed to measure activity of sodium channels using ipsilateral L3/L4/L5 dorsal root ganglion(DRG) cells isolated from the SNI rats.In addition, HEK 293 cells expressing hNav1.7 and hTRPA1 channels were used for measuring the changes in channel activities with or without geraniol by whole-cell patch clamp.Results Geraniol had a fast analgesic effect on hypersensitivity of mechanical pain in the SNI model.It significantly inhibited sodium channels on DRGs isolated from SNI rats and hNav1.7 but not hTRPA1 channels expressed by HEK293 cells.However, high concentrations of geraniol facilitated the activation of HTRPA1 channel stimulated by AITC.Conclusion Geraniol may abirritate hypersensitivity of mechanical pain in the SNI model by specifically inhibiting Nav1.7 channel activity on the DRG cells.

AIM:To investigate the role of caveolin-1 on epithelial-mesenchymal transition (EMT) in human bronchial epithelial (HBE) cells induced by transforming growth factor beta 1 (TGF-β1).METHODS:Immunofluorescence, real-time PCR and Western blot were applied to detect the mRNA and the protein expression of caveolin-1 in the 16HBE cells during EMT.The influence of siRNA-mediated silencing of caveolin-1 on EMT in the 16HBE cells was detected by Western blot.RESULTS:Caveolin-1 was widely present on the cell membrane of the 16HBE cells.The expression of caveolin-1 at mRNA and protein levels was significantly decreased in a time-dependent manner in the 16HBE cells compared with control group (P<0.05) after stimulation with TGF-β1.The morphologic changes of the 16HBE cells induced by TGF-β1 were promoted by caveolin-1 silencing compared with TGF-β1 group.The protein expression of E-cadherin and α-SMA induced by TGF-β1 was promoted by caveolin-1 silencing compared with TGF-β1 group (P<0.05).The phosphorylation levels of AKT and Smad3 were the highest at 30 min and increased significantly compared with control group (P<0.05) after stimulated with TGF-β1.Treatment of the 16HBE cells with TGF-β1 for 30 min after silencing caveolin-1 gene for 24 h significantly increased the phosphorylation levels of AKT and Smad3 compared with TGF-β1 group (P<0.05).CONCLUSION:TGF-β1 down-regulates the expression of caveolin-1 in the 16HBE cells.Caveolin-1 may participate in TGF-β1/Smad pathway and PI3K-AKT pathway, which are the signal transduction pathways for TGF-β1 inducing EMT.