Aim To express and purify recombinant hCGH-CTP fusion protein in high-density suspension culture of Chinese hamster ovary cells (CHO-S), and to verify the lipid accumulation effect of rhCGH-CTP on 3T3-L1 mature adipocytes. Methods The recombinant protein expression vector (pcDNA3. 1-rhCGH-CTP) was constructed, achieved by fusing the human glycoprotein hormone beta 5/alpha 2 cDNA with CTP Linker. The expression plasmid was transiently transfected into the suspended CHO-S to express rhCGH-CTP protein and then purified, and the protein biological activity was verified. Intervention with 3T3-L1 mature adipocyte cells for 24 h was performed to detect the changes of intracellular triglyceride (TG) level. Results Western blot results showed that rhCGH-CTP protein was successfully expressed in CHO-S cells, and the yield was up to 715. 4 mg • L~ . The secreted protein was purified by AKTA pure system with higher purity that was up to 90% as identified by SDS-PAGE. In addition, the intracellular cAMP content of mature adipocytes with high expression of TSHR gene significantly increased after intervention with different concentrations of rhCGH-CTP protein by ELISA kit, indicating that rhCGH-CTP protein had biological activity. Oil red 0 staining showed that compared with the control group, the lipid content of mature adipocytes in the intervention groups with different concentrations of rhCGH-CTP protein significantly decreased (P < 0. 05) . Conclusions The rhCGH-CTP protein has been successfully expressed and purified with biological activity, and effectively reduce TG. This research provides an important theoretical basis for further revealing the physiological role of CGH protein and its potential application in clinical practice.

Through a compound induction method, combined with neurobehavioral, macroscopic characterization and objective pathological evaluation indicators, a murine depression model of liver depression transforming into fire syndrome was constructed and confirmed. The model was constructed using a combination of sleep deprivation, light exposure, and alternate-day food deprivation. Evaluation was conducted at three levels: face validity, constructs validity, and predictive validity. The establishment of the liver depression transforming into fire syndrome depression model was further validated through the counterproof of traditional Chinese medicine formulas. In terms of face validity, compared to the control group, mice in the model group exhibited typical depressive symptoms in neurobehavioral assessments; the general observation of the model group mice reveals disheveled and lackluster fur, along with delayed and easily agitated responses. Additionally, there is a substantial increase in water consumption. In the sleep phase detection of mouse, the model group showed a significant increase in the proportion of time spent in the wake phase during sleep, accompanied by a significant decrease in the proportions of time spent in both non-rapid eye movement (NREM) and rapid eye movement (REM) sleep phases. There are significant differences in physiological indicators such as average blood flow velocity, blood flow rate, tongue, urine, and claw color (r values) in the internal carotid artery. Structural validity demonstrated that levels of 5-hydroxytryptamine (5-HT), dopamine (DA), and γ-aminobutyric acid (GABA) in the hippocampus of model mice decreased significantly, while acetylcholine (ACh), tryptophan (Try), and glutamic acid (Glu) levels increased significantly. Treatment with Danzhi Xiaoyao San led to varying degrees of restoration in the aforementioned neurotransmitters. In terms of predictive validity, the antidepressant paroxetine effectively ameliorated depressive behaviors in the model mice, and the classic formula Danzhi Xiaoyao San demonstrated varying degrees of improvement in depression-related indicators. In conclusion, this study has established a novel animal model of liver-stagnation with the fire depression, thereby expanding the repertoire of traditional Chinese medicine depression models. This development contributes to the diversification of melancholic syndrome models in Chinese medicine, offering a broader spectrum of options for scientific exploration, efficacy evaluation, and drug screening in the future prevention and treatment of depression with traditional Chinese medicine. Animal experiments were conducted with the approval and supervision of the Animal Ethics Committee of Jiangxi University of Traditional Chinese Medicine (ethics number: 20230313040).

Objective This study aimed to investigate the potential relationship between urinary metals copper (Cu), arsenic (As), strontium (Sr), barium (Ba), iron (Fe), lead (Pb) and manganese (Mn) and grip strength. Methods We used linear regression models, quantile g-computation and Bayesian kernel machine regression (BKMR) to assess the relationship between metals and grip strength.Results In the multimetal linear regression, Cu (β=-2.119), As (β=-1.318), Sr (β=-2.480), Ba (β=0.781), Fe (β= 1.130) and Mn (β=-0.404) were significantly correlated with grip strength (P < 0.05). The results of the quantile g-computation showed that the risk of occurrence of grip strength reduction was -1.007 (95％ confidence interval:-1.362, -0.652; P < 0.001) when each quartile of the mixture of the seven metals was increased. Bayesian kernel function regression model analysis showed that mixtures of the seven metals had a negative overall effect on grip strength, with Cu, As and Sr being negatively associated with grip strength levels. In the total population, potential interactions were observed between As and Mn and between Cu and Mn (Pinteractions of 0.003 and 0.018, respectively).Conclusion In summary, this study suggests that combined exposure to metal mixtures is negatively associated with grip strength. Cu, Sr and As were negatively correlated with grip strength levels, and there were potential interactions between As and Mn and between Cu and Mn.

Endodontic diseases are a kind of chronic infectious oral disease.Common endodontic treatment concepts are based on the removal of inflamed or necrotic pulp tissue and the replacement by gutta-percha.However,it is very essential for endodontic treatment to debride the root canal system and prevent the root canal system from bacterial reinfection after root canal therapy(RCT).Recent research,encompassing bacterial etiology and advanced imaging techniques,contributes to our understanding of the root canal system's anatomy intricacies and the technique sensitivity of RCT.Success in RCT hinges on factors like patients,infection severity,root canal anatomy,and treatment techniques.Therefore,improving disease management is a key issue to combat endodontic diseases and cure periapical lesions.The clinical difficulty assessment system of RCT is established based on patient conditions,tooth conditions,root canal configuration,and root canal needing retreatment,and emphasizes pre-treatment risk assessment for optimal outcomes.The findings suggest that the presence of risk factors may correlate with the challenge of achieving the high standard required for RCT.These insights contribute not only to improve education but also aid practitioners in treatment planning and referral decision-making within the field of endodontics.

Objective To analyze the characteristics and differences of gut flora in patients with osteoporosis of different traditional Chinese medicine(TCM)syndrome types,thus to find out new therapeutic targets for the treatment of osteoporosis.Methods The clinical data and fecal samples of 57 patients with osteoporosis recruited in the Department of Osteoporosis,the Third Affiliated Hospital of Guangzhou University of Chinese Medicine from February 2023 to November 2023 were collected.The patients were differentiated as the TCM syndrome types of spleen and kidney yang deficiency group(22 cases),liver and kidney yin deficiency group(18 cases)and kidney deficiency and blood stasis group(7 cases).The differences in the structure and abundance of gut flora in patients with the above three TCM syndrome types were analyzed by16S rDNA sequencing technique.Results The results of Alpha(α)diversity showed that there were no significant differences in species richness,evenness and diversity of gut flora among the three groups(P＞0.05).Beta(β)diversity analysis showed that there were significant differences in gut flora among the three groups(P=0.011).At the phylum level,Bacteroidota,Firmicutes and Proteobacteria were the top three kinds of gut flora with the highest proportion in the three groups.At the genus level,the relative abundance of Bacteroides,Escherichia-Shigella,and Faecalibacterium was higher in the group of liver and kidney yin deficiency and in the group of kidney deficiency and blood stasis,while the group of spleen and kidney yang deficiency has a large proportion of other bacteria.Conclusion There exist differences in the composition of gut flora among patients with different TCM syndrome types,and the differences are shown at the level of phylum and genus.The results indicate that gut flora may be the targets in the TCM prevention and treatment of osteoporosis.

Objective To measure the distance between the lateral compression Ⅱ(LC-Ⅱ)screw guide needle and the surrounding important structures around the anterior inferior iliac spine in pelvic fractures and to locate the needle point,so as to provide anatomical reference for clinical nail placement.Methods Totally 40 adult gross specimens of embalming were implanted with LC-Ⅱ screw guide needle under the surveillance of C-arm machine,and the specimens were dissected.The shortest distance between the insertion point and the lateral femoral cutaneous nerve,femoral nerve,femoral artery,femoral vein,anterior superior iliac spine and inguinal ligament was measured.The triangle was constructed between the insertion point,anterior superior iliac spine and inguinal ligament,and the exact location of the entry point was calculated.Results The average distance between the insertion point of the male needle and the femoral vein was(50.67±7.29)mm＞the anterior superior iliac spine(43.83±7.58)mm＞the femoral artery(38.35±6.63)mm＞the femoral nerve(31.17±1.67)mm=the inguinal ligament(28.69±6.59)mm＞the lateral femoral cutaneous nerve(7.98±3.81)mm.The mean distance between the insertion point of the female needle and the anterior superior iliac spine was(45.28±7.07)mm=femoral vein(43.72±6.89)mm＞femoral artery(33.76±6.33)mm＞femoral nerve(25.66±6.46)mm=inguinal ligament(23.22±5.00)mm＞lateral femoral cutaneous nerve(8.97±4.76)mm.The projection distance of the entry point was 31.77 mm for men and 38.41 mm for women.The Angle b was 42.81°for men and 31.71° for women.Conclusion The lateral femoral cutaneous nerve is most vulnerable to injury when LC-Ⅱ screw is inserted,and the risk of injury has nothing to do with sex.The insertion point positioning method a and b made LC-Ⅱ screw placement quickly,safely and accurately,and reduced fluoroscopy time and frequency.

Objective To investigate the prenatal diagnosis and perinatal outcomes between anterior pla-centa accreta and non-anterior placenta.Methods A retrospective analysis was done for 560 pregnant women who were diagnosed with placenta accreta and delivered in the Third Affiliated Hospital of Guangzhou Medical Uni-versity.According to the location of the placenta,the group was dividing into anterior placenta accreta group(319 cases)and non-anterior placenta accreta group(241 cases).The general characteristics,maternal and infant out-comes of the two groups were analyzed.The non-anterior placenta accrete group(241 cases)then were dividing into two groups according to the time of clear diagnosis.Those who were firstly diagnosed with placenta accrete dur-ing or after the operation was the intrapartum diagnosis group(missed diagnosis)(70 cases),and those who were diagnosed with clear placenta accreta before the delivery was prenatal diagnosis group(171 cases).The general characteristics,maternal and infant outcomes of the two groups were also analyzed.Results There were statisti-cally significant differences in the parity,history of cesarean section,delivery mode,degree of placenta accreta,missed diagnosis rate,neonatal birth weight,and hysterectomy rate between the non-anterior placenta accrete group and the anterior placenta accreta group.In the case of prenatal diagnosis of different degrees of placenta accreta,the prenatal diagnosis rate of placental adhesion in the non-anterior placenta accreta group was lower than that of the anterior placenta accreta group,which was statistically significant.In the non-anterior placenta accrete group,there were statistically significant differences in the age,cesarean section history,placenta previa status,mode of delivery,degree of implantation,24-hour bleeding volume,blood transfusions,NICU transfer rate,uterine loss rate between the intrapartum diagnosis group(missed diagnosis)and the prenatal diagnosis group.Conclusions The high-risk factors of patients with non-anterior placenta accreta are different from those of patients with anterior placenta accreta.Multiple births and a history of cesarean section are high-risk factors for anterior placenta accreta patients.Non-anterior placenta accreta are more likely to be missed diagnosed,especially the placental adhesion.For pregnant women with non-anterior placenta accreta missed diagnosis,there is a high rate of adverse birth out-comes,especially in the rate of neonatal transfer to the NICU.

Objective To investigate the risk factors for bronchopulmonary dysplasia(BPD)in twin preterm infants with a gestational age of<34 weeks,and to provide a basis for early identification of BPD in twin preterm infants in clinical practice.Methods A retrospective analysis was performed for the twin preterm infants with a gestational age of<34 weeks who were admitted to 22 hospitals nationwide from January 2018 to December 2020.According to their conditions,they were divided into group A(both twins had BPD),group B(only one twin had BPD),and group C(neither twin had BPD).The risk factors for BPD in twin preterm infants were analyzed.Further analysis was conducted on group B to investigate the postnatal risk factors for BPD within twins.Results A total of 904 pairs of twins with a gestational age of<34 weeks were included in this study.The multivariate logistic regression analysis showed that compared with group C,birth weight discordance of>25%between the twins was an independent risk factor for BPD in one of the twins(OR=3.370,95%CI:1.500-7.568,P<0.05),and high gestational age at birth was a protective factor against BPD(P<0.05).The conditional logistic regression analysis of group B showed that small-for-gestational-age(SGA)birth was an independent risk factor for BPD in individual twins(OR=5.017,95%CI:1.040-24.190,P<0.05).Conclusions The development of BPD in twin preterm infants is associated with gestational age,birth weight discordance between the twins,and SGA birth.

Objective To observe the clinical effect of arthroscopic repair with metal suture anchor on type Ⅱ superior labral anterior to posterior(SLAP)lesion of the youth.Methods The clinical data of 32 young patients with type Ⅱ SLAP lesion who underwent arthroscopic repair with metal suture anchor were retrospectively analyzed.The postoperative complications of patients were counted.The visual analogue scale(VAS)score was used to evaluate the pain of patients.Rowe shoulder scoring system,Constant-Murley scoring system and American Shoulder and Elbow Surgeons(ASES)scoring system were used to evaluate the shoulder joint function.Results All the 32 patients successfully completed the operation,and they were followed up for 12 to 27 months,with the mean follow-up time of 15 months.After operation,the symptoms of shoulder pain and limited movement of 32 patients were significantly relieved,without complications such as shoulder joint infection,shoulder joint instability,or neurovascular injury.The postoperative VAS score,Rowe score,Constant-Murley score and ASES score of all patients were better than those before operation(P<0.05).Conclusion Arthroscopic repair with metal suture anchor on type Ⅱ SLAP lesion of the youth can achieve satisfactory clinical outcomes.

Objective:To analyze thalassemia genotypes and distribution of children in Wuzhou Guangxi,and evaluate the diagnostic value of HbA2 in children's thalassemia screening,so as to provide scientific evidence for the prevention and control strategies of thalassemia.Methods:Four hundred and fifty-eight children suspected with thalassemia in Wuzhou were enrolled from March 2017 to June 2022.The level of HbA2 was detected using Bio-Rad VARIANT Ⅱ Hb analysis system.The deletion of α-thalassemia was measured with gap-PCR assay,and the point mutation of α-and β-thalassemia was tested with DNA reverse dot blot hybridization assay.The receiver operating characteristic(ROC)curve was used to analyze the diagnostic value of HbA2 for children's thalassemia.Results:A total of 304 thalassemia carriers were detected in 458 children,accounting for 66.38％.One hundred and seventy-five cases were defined to be α-thalassemia,with the main type of--SEA/αα(54.86％).Thirty-six cases were defined to be intermediate α-thalassemia,with the main type of-α3.7/--SEA(9.72％).In 108 cases with β-thalassemia,βCD41-42/βN was the main type,accounting for 49.07％,followed by βIVS-Ⅱ-654/βN(14.81％).Seven cases were moderate/severe β-thalassemia(predominantly β-28/β-28 and βCD41-42/βCD17)Twenty-one genotypes of α-and β-thalassemia were found in the children.There was significant difference of HbA2 level between the children with different types of thalassemia and healthy controls(all P＜0.001).ROC curve analysis showed that the sensitivities of HbA2 for α-thalassemia,β-thalassemia and αβ-thalassemia were 74.3％,82.4％and 85.7％,with the optimal cut-off values of 2.60％,3.60％and 3.70％,respectively,the specificities were 64.3％,96.1％and 96.8％,and the area under the curve were 0.690,0.887 and 0.916,respectively.Conclusion:The thalassemia genotypes of children in Wuzhou are diverse.It is necessary to further strengthen the prevention and control measure of thalassemia to reduce birth defects and improve birth quality.