ObjectiveTo explore the mechanisms associated with Mahoniae Caulis alkaloids （MA） in ameliorating depression by network pharmacology， molecular docking， and animal experiments. MethodsThe component targets of MA were obtained through Swiss Target Prediction and TCMIP database. The depression targets were collected through TCMIP， Genecards， HPO， DrugBank and OMIM database. The depression targets were collected through TCMIP， Genecards， HPO， DrugBank and OMIM database. Protein-protein interaction （PPI） network was constructed by protein interaction analysis （STRING） database. Gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） analyses were performed through Bioinformatics （DAVID） database. The docking of components and targets was performed by AGFR. The mouse model of depression was established by intraperitoneal injection of corticosterone （CORT） once a day for 35 consecutive days. Sixty mice were randomly allocated into control （0.9% normal saline）， model （CORT， 20 mg·kg^-1）， positive control （fluoxetine hydrochloride， 3.6 mg·kg^-1）， and MA （10， 5， and 2.5 mg·kg^-1） groups. Each group was administrated with corresponding medicine or normal saline once a day for 28 consecutive days. The depression-like behavior of mice was observed. The pathological changes of prefrontal cortex in mice were observed by hematoxylin-eosin staining. Terminal deoxynucleotidyl dUTP transferase nick end labeling （TUNEL） was employed to observe the apoptosis of neurons in the prefrontal cortex. Enzyme-linked immunosorbent assay was employed to assess the serum levels of brain-derived neurotrophic factor （BDNF）， dopamine （DA）， 5-hydroxytryptamine （5-HT）， and norepinephrine （NE） in mice. The mRNA levels of cyclic adenosine monophosphate （cAMP） pathway-related factors and inflammatory factors were determined by Real-time PCR. Western blot was employed to determine the expression of cAMP pathway-related factors and connexin 43 （Cx43）. ResultsA total of 434 component targets and 545 depression targets were obtained， including 84 common targets， among which 10 core targets were screened out. GO analysis predicted 34 biological processes， 15 cell components， and 11 molecular functions. The KEGG pathways were mainly related to gap junction and cAMP signaling pathway. The core components had good binding affinity with the core targets. The results of animal experiments showed that compared with the control group， CORT prolonged the immobility time of mice in forced swimming and tail suspension tests （P<0.01）， lowered the serum levels of NE， BDNF， and 5-HT （P<0.05）， up-regulated the mRNA levels of nuclear factor-κB （NF-κB） and interleukin-6 （IL-6） in the brain tissue （P<0.05）， and down-regulated the mRNA levels of cyclic adenosine monophosphate effector binding protein （CREB） and BDNF （P<0.05） and the protein levels of protein kinase （PRKACA）， phosphorylation （p）-CREB/CREB， BDNF， and Cx43 （P<0.05） in the brain tissue. Compared with the model group， high-dose MA reduced the immobility time of mice in forced swimming （P<0.05） and tail suspension （P<0.01） tests， raised the serum levels of NE， BDNF， and 5-HT （P<0.01）， down-regulated the mRNA level of NF-κB （P<0.01）， and up-regulated the mRNA level of BDNF （P<0.01） and protein levels of PRKACA， p-CREB/CREB， BDNF， and Cx43 （P<0.05）. ConclusionMA alleviates the CORT-induced depressive behavior of mice. It may play an antidepressant role by regulating cAMP signaling pathway and gap junction pathway， improving synaptic plasticity and gap junction function， and reducing neuroinflammation.

ObjectiveTo explore the mechanisms associated with Mahoniae Caulis alkaloids （MA） in ameliorating depression by network pharmacology， molecular docking， and animal experiments. MethodsThe component targets of MA were obtained through Swiss Target Prediction and TCMIP database. The depression targets were collected through TCMIP， Genecards， HPO， DrugBank and OMIM database. The depression targets were collected through TCMIP， Genecards， HPO， DrugBank and OMIM database. Protein-protein interaction （PPI） network was constructed by protein interaction analysis （STRING） database. Gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） analyses were performed through Bioinformatics （DAVID） database. The docking of components and targets was performed by AGFR. The mouse model of depression was established by intraperitoneal injection of corticosterone （CORT） once a day for 35 consecutive days. Sixty mice were randomly allocated into control （0.9% normal saline）， model （CORT， 20 mg·kg^-1）， positive control （fluoxetine hydrochloride， 3.6 mg·kg^-1）， and MA （10， 5， and 2.5 mg·kg^-1） groups. Each group was administrated with corresponding medicine or normal saline once a day for 28 consecutive days. The depression-like behavior of mice was observed. The pathological changes of prefrontal cortex in mice were observed by hematoxylin-eosin staining. Terminal deoxynucleotidyl dUTP transferase nick end labeling （TUNEL） was employed to observe the apoptosis of neurons in the prefrontal cortex. Enzyme-linked immunosorbent assay was employed to assess the serum levels of brain-derived neurotrophic factor （BDNF）， dopamine （DA）， 5-hydroxytryptamine （5-HT）， and norepinephrine （NE） in mice. The mRNA levels of cyclic adenosine monophosphate （cAMP） pathway-related factors and inflammatory factors were determined by Real-time PCR. Western blot was employed to determine the expression of cAMP pathway-related factors and connexin 43 （Cx43）. ResultsA total of 434 component targets and 545 depression targets were obtained， including 84 common targets， among which 10 core targets were screened out. GO analysis predicted 34 biological processes， 15 cell components， and 11 molecular functions. The KEGG pathways were mainly related to gap junction and cAMP signaling pathway. The core components had good binding affinity with the core targets. The results of animal experiments showed that compared with the control group， CORT prolonged the immobility time of mice in forced swimming and tail suspension tests （P<0.01）， lowered the serum levels of NE， BDNF， and 5-HT （P<0.05）， up-regulated the mRNA levels of nuclear factor-κB （NF-κB） and interleukin-6 （IL-6） in the brain tissue （P<0.05）， and down-regulated the mRNA levels of cyclic adenosine monophosphate effector binding protein （CREB） and BDNF （P<0.05） and the protein levels of protein kinase （PRKACA）， phosphorylation （p）-CREB/CREB， BDNF， and Cx43 （P<0.05） in the brain tissue. Compared with the model group， high-dose MA reduced the immobility time of mice in forced swimming （P<0.05） and tail suspension （P<0.01） tests， raised the serum levels of NE， BDNF， and 5-HT （P<0.01）， down-regulated the mRNA level of NF-κB （P<0.01）， and up-regulated the mRNA level of BDNF （P<0.01） and protein levels of PRKACA， p-CREB/CREB， BDNF， and Cx43 （P<0.05）. ConclusionMA alleviates the CORT-induced depressive behavior of mice. It may play an antidepressant role by regulating cAMP signaling pathway and gap junction pathway， improving synaptic plasticity and gap junction function， and reducing neuroinflammation.

Tick-borne encephalitis is a central nervous system disease caused by infections with tick-borne pathogens, which is characterized by severe clinical symptoms, multiple sequelae, and a high fatality rate. Currently, there is no cure for tick-borne encephalitis. Tick-borne encephalitis virus (TBEV) is the most common pathogen of tick-borne encephalitis. Therefore, rapid and accurate detection of TBEV contributes to reducing the mortality of tick-borne encephalitis, improving patients' prognosis, and reducing the risk of TBEV transmission. The currently available serological tests for detection of TBEV infections mainly include neutralization test, enzyme-linked immunosorbent assay (ELISA), immunofluorescence assay, and nucleic acid tests mainly include polymerase chain reaction (PCR), loop-mediated isothermal amplification (LAMP), reverse transcription polymerase spiral reaction, clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated proteins (Cas)-based assays. This review summarizes the progress of researches on serological and nucleic acid tests for detection of TBEV infections, so as to provide insights into prevention and control of tick-borne encephalitis.

Biomarkers is of great significance for clinical diagnosis. Lateral flow assay (LFA) is a test method which developed based on paper chromatography and has a wide range of applications in testing disease biomarkers. The current researches of LFA mainly focus on the detection specificity, precision and sensitivity, as well as multiple detection of disease markers or simultaneous detection of multiple diseases. This artical reviews the systemic design, working principles of LFA along with the application research of LFA in detecting disease markers, in order to provide reference for development and application of LFA.

Objective:To explore the epileptic phenotype spectrum of SLC6A1 gene mutations and their genotype-phenotype correlation. Methods:Four hundred patients with epilepsy of unknown etiology admitted to Epilepsy Center, Department of Neurology, Second Affiliated Hospital of Guangzhou Medical University from July 2019 to July 2024 were enrolled to screen the SLC6A1 gene mutations; the clinical characteristics, mutation pathogenicity, and changes of hydrogen bond between amino acids, stability and amino acid hydrophobicity of SLC6A1 gene encoded proteins caused by missense mutations in patients with SLC6A1 gene mutations were analyzed. At the same time, a comprehensive search was conducted in PubMed, HGMD and CNKI databases to collect the publicly reported SLC6A1 gene mutations related to epilepsy up to September 8, 2024; differences in proportion of missense mutations between the two most common and featured epileptic phenotypes and proportion of missense mutations in loops of SLC6A1 gene coding proteins were analyzed. Results:Five patients carried SLC6A1 gene mutations in 400 patients with epilepsy of unknown etiology: 2 had de novo heterozygous canonical splice site mutations (c.850-1G>A and c.1324-1G>A), with phenotypes as partial epilepsy combined with severe development delay and childhood absence epilepsy combined with mild developmental delay; 2 had de novo heterozygous missense mutations (c.187G>A/p.Gly63Ser and c.1081C>A/p.Pro361Thr), with phenotypes as partial epilepsy combined with mild development delay and generalized epilepsy combined with severe development delay; and one had heterozygous missense mutation of unknown origin (c.700G>A/p.Gly234Ser), with phenotype as Lennox-Gastaut syndrome. Four de novo mutations were evaluated as having pathogenic or likely pathogenic features, and one mutation of unknown origin was evaluated as of uncertain significance. In addition, 3 missense mutations caused significant changes in number or bonding form of hydrogen bonds between amino acids of the encoded proteins, with obviously decreased stability and hydrophobicity of the encoded proteins. (2) Results of literature analysis showed that 84 SLC6A1 mutations have been reported to be associated with epilepsy; combined with the genetic results in this study, a total of 89 SLC6A1 mutations were identified, including 53 missense mutations, 33 nonsense mutations, and 3 in-frame/in-del mutations; 7 epilepsy phenotypes were involved, including 38 patients with myoclonic atonic epilepsy (MAE), 16 with epilepsy, 12 with epileptic encephalopathy, 8 with childhood absence epilepsy, 6 with childhood-onset epilepsy, 6 with generalized epilepsy, and 3 with focal epilepsy. No significant difference in proportion of missense mutations was noted between MAE and epileptic encephalopathy patients ( P>0.05); however, the proportion of missense mutations in loops of the epileptic encephalopathy patients was significantly higher than that of the MAE patients ( P<0.05). Conclusion:SLC6A1 gene mutations can cause complex and diverse epilepsy phenotype spectrum, and most patients are accompanied by developmental delay; subregional effect of the encoded protein molecules may be a potential mechanism for different clinical phenotypes between MAE and epileptic encephalopathy caused by SLC6A1 gene mutations.

Objective:To investigate the strategies and clinical effects of free tissue flaps in repairing massive destructive burn wounds.Methods:This study was a retrospective observational study. From June 2014 to October 2023, 51 burn patients with 53 massive destructive burn wounds which met the inclusion criteria were admitted to Tongren Hospital of Wuhan University & Wuhan Third Hospital, including 47 males and 4 females, aged 21 to 77 years. After debridement, the wound area needed to be repaired by tissue flaps ranged from 20.0 cm×12.5 cm to 50.0 cm×15.0 cm. Emergency, early, or delayed transplantation of tissue flaps was performed to repair the wounds. Six head, face, and neck wounds, 5 upper limb wounds, and 1 lower limb wound were repaired with latissimus dorsi myocutaneous flaps. Eleven upper limb wounds and 6 lower limb wounds were repaired with anterolateral thigh flaps. Seven upper limb wounds, 5 lower limb wounds, and 2 face and neck wounds were repaired with paraumbilical perforator flaps. One lower limb wound was repaired with lower abdominal flap. Two face and neck wounds, 2 upper limb wounds, and 1 lower limb wound were repaired with inguinal flaps. One upper limb wound was repaired with scapular flap. One trunk wound was repaired with the "fillet flap" from the abandoned upper limb after amputation. Three head, face, and neck wounds, 3 lower limb wounds, and 1 upper limb wound were repaired with omental flaps. Four wounds were repaired by combined transplantation of the above-mentioned tissue flaps. Three wounds were repaired by fractional transplantation of the above-mentioned tissue flaps. The size of a single harvested tissue flap ranged from 15.0 cm×5.0 cm to 45.0 cm×25.0 cm. The arterial and venous anastomoses were added as the circumstances might require to improve tissue flap circulation. The wounds in the flap donor sites were sutured directly or repaired by split-thickness skin grafts from head. The general condition of patients during treatment was recorded. After surgery, the survival of tissue flaps, the healing of wounds in the recipient sites, limb salvage, the healing of wounds in the flap donor sites, and the survival of skin grafts were observed. The healing of wounds in the recipient sites was observed during follow-up. At the last follow-up, the upper limb function after successful limb salvage was evaluated by the Arm, Shoulder, and Hand Disability Scoring Scale, the weight-bearing walking ability of patients with successful lower limb salvage was evaluated by the Holden walking ability classification, and the patient's satisfaction with the efficacy was assessed by 5-grade Likert scale.Results:During treatment, one case with electrical burn of the head suffered from aggravated cerebral edema, cerebral hernia and coma recurred. After dehydration, hormone therapy, and so on, the patient recovered and incomplete hemiplegia on the right limb was left. The shock symptoms of 4 patients got worse, which were gradually controlled after anti-shock with fluid supplement and colloid injection. One patient developed hemorrhagic shock after tissue flap transplantation, which was corrected by fluid infusion and red blood cell suspension transfusion. Vascular crisis occurred in 3 tissue flaps after surgery, of which 2 survived and 1 eventually became necrotic after exploration. Partial necrosis occurred in 2 tissue flaps and slight necrosis occurred at the edge of 5 tissue flaps, which all healed after tissue flap trimming, vacuum sealing drainage (VSD), and skin grafting treatment. All the other tissue flaps survived. There were 12 recipient wounds with residual necrotic tissue after surgery, which healed after debridement, VSD, and skin grafting treatment. The remaining 41 wounds healed well. Among the 40 limb wounds of 39 patients, 5 patients had 3 upper limb and 2 lower limb amputations. The remaining 35 limbs of 34 patients were successfully salvaged. The donor site wounds healed well, and the skin grafts survived well. During the follow-up of 6 to 52 months, all the head, face, neck, and trunk wounds were well repaired. The fester recurred in 7 limb wounds 3 to 9 months after surgery and healed after debridement, drainage, and tissue flap repair. At the last follow-up, the functional scores of 22 upper limbs after successful limb salvage ranged from 0 to 100 (with an average of 50), and the Holden walking ability classification of 12 patients with 13 lower limbs after successful limb salvage was level Ⅴ in 8 cases, level Ⅳ in 2 cases, and level Ⅲ in 2 cases. The patients were very satisfied with the efficacy in 33 cases, relatively satisfied in 12 cases, and not quite satisfied in 6 cases.Conclusions:Systematic evaluation and treatment of the whole body condition, choosing the appropriate repair time, and using appropriate tissue flaps to repair massive destructive burn wounds will help ensure the safety of treatment, improve the repair effect, and reduce the injury of donor sites, thus optimizing the repair effect.

【Objective】 To investigate the compatibility of human albumin and its internal packaging materials of Sinopharm Lanzhou Biopharmaceutical Co., Ltd. 【Methods】 One batch of inner packaging materials (medium borosilicate glass-molded injection bottle and halogenated butyl rubber plug for injection) was extracted with 4 extraction solvents to conduct the toxicological evaluation of potential inner packaging extracts. Through the simulated acceleration test, the trend analysis of the elements in the sample and the inner surface of the glass bottle were observed, and the routine drug inspection items during the long-term stability test process were determined. 【Results】 The detection results of the leaching elements of the internal packaging materials did not exceed the limit of 50%, and the organic matter safety threshold (margin of safety, MOS) was greater than 1.0, indicating that both the leaching elements and the organic matter had no safety risk to the user under the current exposure. The results of the simulated acceleration test show that the drug will not have the risk of peeling tablets after the long-term stability condition was placed for a period of time, and the routine inspection items of the long-term stability test drugs all meet the requirements of the pharmacopoeia. 【Conclusion】 The inner packaging material has no significant impact on the quality of drugs and has good overall compatibility, making it suitable for packaging human albumin.

This article reports a case of oligomenorrhea accompanied by acne over several years. The patient presented with hyperandrogenism and insulin resistance. The initial diagnosis considered polycystic ovary syndrome(PCOS) and was treated with oral medication for 2 months, with no significant improvement. Further ultrasound examination revealed a right ovarian heterogeneous mass (3.2 cm×3.0 cm). The patient underwent laparoscopic resection of the right ovarian lesion. Postoperative pathology revealed ovarian sex cord-stromal cell tumor, adult granulosa cell tumor(diffuse type). Testosterone levels decreased to the normal 3 days after the surgery, and menstruation resumed within 2 months. This article summarizes the presented case and reviews the relevant literature. For women with severe hyperandrogenism and masculinization, it is recommended to thoroughly assess the possibility of androgen-secreting tumors.

Objective:To investigate the clinical effects of free transplantation of inguinal flap in repairing electrical burn wounds of mice in the limbs.Methods:A retrospective observational study was conducted. From January 2012 to May 2023, 24 male patients with electrical burns in the limbs meeting the inclusion criteria, aged 20 to 69 years, were admitted to the Department of Burns of Tongren Hospital of Wuhan University & Wuhan Third Hospital. Totally 27 limbs were involved, including 18 upper limbs and 9 lower limbs. After debridement, the wound area in the limbs proposed to be repaired with the inguinal flap was 5.0 cm×1.5 cm-20.0 cm×9.0 cm. A total of 26 operations were performed in 24 patients, and 27 free inguinal flaps with area being 5.0 cm×3.5 cm-22.0 cm×12.0 cm were resected, including 6 for lobed transplantation, 2 for chimeric transplantation with iliac bone graft, 3 for conjoined transplantation with pedicled latissimus dorsi myocutaneous flap, and 1 for conjoined transplantation with free paraumbilical perforator flap. Indocyanine green angiography was used to evaluate the blood supply of the inguinal flap during operation, to guide arterial supercharge during transplantation. Among the transplanted inguinal flaps, 5 were pedicled with the common trunk of the superficial circumflex iliac artery and the superficial abdominal wall artery, 13 were pedicled with the superficial circumflex iliac artery alone, 6 were pedicled with the superficial abdominal wall artery alone, and the remaining 3 were simultaneously anastomosed with the superficial circumflex iliac artery and the superficial abdominal wall artery for arterial supercharge. In another patient, the deep branch of the superficial circumflex iliac artery was used as a bridging vessel to recanalize the proper artery of the index finger in the distal segment. The wound in the donor area were sutured directly. The blood supply status of the inguinal flap immediately after resection but before cutting off blood vessels and after transplantation and vascular anastomosis was recorded. The survival of the tissue flap, the fixation of the iliac bone graft, and the healing of secondary defect in the donor area were observed after surgery. The appearance of the inguinal flap and the functional recovery of the recipient area were followed up. At the last follow-up, the function of the affected upper limb was scored using the Arm, Shoulder, and Hand Disability (DASH) scoring scale, and the satisfaction of patients with the efficacy of each surgical treatment was investigated by referring to the 5-level Likert scale.Results:Totally 27 inguinal flaps showed good arterial blood perfusion and venous return immediately after resection but before cutting off blood vessels and after transplantation and vascular anastomosis. Except for one inguinal flap that developed necrosis 12 days after operation and was repaired by split-thickness skin graft from the head, the remaining 26 inguinal flaps and conjointly transplanted tissue flaps survived completely without vascular crisis. The iliac bone graft was fixed securely after operation. All secondary defects in donor area healed well after operation. After 6 to 36 months of follow-up, the inguinal flap was soft in texture, without obvious swelling, the function of the recipient area was significantly improved as compared with that before surgery, and the protective feeling of the inguinal flap was restored. At the last follow-up, the functional scores of DASH scoring scale of the 18 affected upper limbs ranged from 0 to 100 (with a mean of 27). The patients were very satisfied with the efficacy of 17 surgical treatments and relatively satisfied with the efficacy of 9 surgical treatments.Conclusions:The donor area of the inguinal flap is concealed, and the damage resulted from flap resection in this area is small. This flap has a rich blood supply. In the electrical burn wounds of mice in the limbs repaired with lobed transplantation, chimeric transplantation, and conjoined transplantation with other flaps, the appearance and function of the recipient area are well restored, and the patients' satisfaction with the efficacy of surgical treatment is high, which is worthy of clinical promotion.

ObjectiveTo establish a rapid screening method for germplasm materials of Gastrodia elata with high purity， and lay a foundation for pure line breeding and cross breeding. MethodBased on the whole genome sequencing and population resequencing of G. elata， 20 restriction fragment length polymorphism (RFLP) markers were developed by single nucleotide polymorphism (SNP) sites. The polymerase chain reaction (PCR)-RFLP method was used to carry out restriction endonuclease experiments on 20 RFLP markers of 15 G. elata germplasms. According to the number of enzymatic bands at 20 RFLP marker sites, the purity of 15 germplasms was calculated and evaluated. On this basis, genome resequencing technology was used to verify the assessment results. ResultTen germplasm materials with purity greater than 95% were screened out by PCR-RFLP method， 3 of which had 95% purity and 7 had 100% purity. Nine germplasm materials with purity greater than 95% were screened out by genome resequencing methods, and 8 of them were consistent with the results of PCR-RFLP. ConclusionThe PCR-RFLP method established in this study for screening G. elata germplasms with high purity precision of RFLP markers has 80% precision and 89% accuracy. The method is simple， efficient， and significantly less expensive than genome resequencing method, which provides technical support for pure line breeding of G. elata and references for breeding of other Chinese medicinal materials.