BACKGROUND:Stem cell transplantation is a new way to prevent and cure intervertebral disc degeneration.However,whether the transplanted stem cells can survive,proliferate,differentiate,and restore the function of nucleus pulposus cells after transplantation,is the key and difficult point to overcome. OBJECTIVE:To explore the effects of Bushenhuoxue decoction on survival,proliferation,and nucleus pulposus-like differentiation of adipose-derived stem cells. METHODS:A Transwell chamber was used to construct a co-culture model of human adipose-derived stem cells and human degenerative nucleus pulposus cells.The experiment was divided into control group,model group,drug-containing serum group,and drug-free serum group.Except for the control group,the co-culture system of other groups was treated with 50 μmol/L tert-butyl hydrogen peroxide for 24 hours.The drug-containing serum group and drug-free serum group were treated with DMEM low-glucose complete culture medium containing drug-containing serum of Bushenhuoxue decoction or drug-free serum with 20％volume fraction for 48 hours.The sublayer adipose-derived stem cells were taken.Toluidine blue staining was used to detect proteoglycan synthesis levels.Real-time PCR method was used to detect mRNA expression of type Ⅱ collagen,proteoglycan and SRY-box transcription factor 9.The protein expression of SOX9 was detected by western blot assay.Lactate dehydrogenase assay was used to detect cytotoxicity.Flow cytometry was used to detect reactive oxygen species,and β-galactosidase staining was used to detect cell senescence. RESULTS AND CONCLUSION:(1)Compared with the control group,the proportion of necrotic cells in the model group increased;toluidine blue staining became lighter,and the expression levels of type Ⅱ collagen,proteoglycan,SOX9 mRNA and SOX9 protein decreased(P<0.05).Compared with the model group,the drug-containing serum of Bushenhuoxue decoction could significantly reduce cell injury and promote the expression of type Ⅱ collagen,proteoglycan,SOX9 mRNA,and SOX9 protein(P<0.05),but the improvement in the drug-free serum group was not significant(P>0.05).(2)Compared with the control group,the contents of cytotoxicity,reactive oxygen species,and cell senescence in the model group were significantly increased.Compared with the model group,the microenvironment of the coculture system was significantly improved by drug-containing serum of Bushenhuoxue decoction(P<0.05),while drug-free serum had no significant effect on the microenvironment of the co-culture system(P>0.05).(3)The results show that Bushenhuoxue decoction can promote the survival,proliferation,and nucleus pulposus-like differentiation of adipose-derived stem cells.

Persistent Müllerian duct syndrome(PMDS) is a rare disorder that arises from a lack of active anti-Müllerian hormone(AMH) or type Ⅱ AMH receptor(AMHR2) deficiency in males with a normal 46, XY chromosome karyotype.It presents that the external genitalia appears normally while the Müllerian duct structure(uterus, fallopian tubes, upper vagina) persists in the body.Common pathogenic factors are mutations in the AMH and AMHR2 genes, inherited in an autosomal recessive manner.This study reported two families with PMDS.The first patient was diagnosed with PMDS due to cryptorchidism in May 2019.Gene sequencing analysis revealed a new missense mutation(c.579G>T; p.W193C) and a splicing mutation(c.622-3C>A; splicing) in the AMHR2 gene.His father had the missense mutation(c.579G>T; p.W193C), and his mother had the splicing mutation(c.622-3C>A; splicing).The second patient was diagnosed with PMDS due to bilateral cryptorchidism, transverse testis ectopia in the right testicle in March 2023.Undegraded Müllerian tube derivatives were found between the two testicles, and serum AMH levels were very high(565.00 μg/L).Gene sequencing analysis reported that the AMHR2 gene had a new deletion mutation(c.835_837del; p.Leu279del).Both his father and mother had a deletion mutation(c.835_837del; p.Leu279del).This study reports two new AMHR2 gene mutations that expand the mutation sites of this rare disease.It is recommended to consider PMDS in the differential diagnosis of cryptorchidism, undergo surgery as early as possible, and treat Müllerian duct derivatives based on individual anatomical characteristics.

Objective:To investigate the efficacy and safety of AcoStream peripheral thrombus aspiration system combined with catheter-directed thrombolysis in the treatment of lower extremity deep vein thrombosis.Methods:The clinical data of 16 lower extremity deep vein thrombosis cases treated with AcoStream peripheral thrombus aspiration system combined with catheter-directed thrombolysis, admitted to the authors′ hospital from May 2022 to November 2022, were retrospectively analyzed. The differences in circumferential diameter between the affected limb and the healthy side, venous patency score, thrombus clearance grade and intraoperative blood loss were observed and compared. The Villalta score was used during the follow up. Paired sample t-test and Wilcoxon rank sum test were used to compare the changes in the observed indicators before and after treatment to evaluate the efficacy. Results:Treatment were successfully performed in all patients. Before treatment, the circumference differences between the affected and unaffected thighs and calves were (3.69±0.97) and (3.34±0.75)cm, respectively, the venous patency score of the affected side was 8(7.25,9) points. After treatment, the circumference differences between the affected and unaffected thighs and calves were (0.81±0.68) and (0.84±0.70)cm, respectively. The venous patency score of the affected side was 1(0,1)points, with statistically significant differences ( P<0.001). Grade Ⅲ thrombus clearance was achieved in 7 patients, grade Ⅱ thrombus clearance was achieved in 9 patients. The average blood loss during thrombus aspiration was (133.1±12.0) ml. Following up for 6 months, the Villalta score was 0(0,1.75) points. Conclusion:Acostream peripheral thrombus aspiration system combined with catheter-directed thrombolysis is safe and effective for the treatment of lower extremity deep venous thrombosis, with satisfactory short-term efficacy and high clinical application value.

Yeast autolysis affects the flavor and quality of beer. The regulation of yeast autolysis is a need for industrial beer production. Previous studies on brewer's yeast autolysis showed that the citric acid cycle-related genes had a great influence on yeast autolysis. To explore the contribution of isocitrate dehydrogenase genes in autolysis, the IDP1 and IDP2 genes were destroyed or overexpressed in typical lager yeast Pilsner. The destruction of IDP1 gene improved the anti-autolytic ability of yeast, and the anti-autolytic index after 96 h autolysis was 8.40, 1.5 times higher than that of the original strain. The destruction of IDP1 gene increased the supply of nicotinamide adenine dinucleotide phosphate (NADPH) and the NADPH/NADP⁺ ratio was 1.94. After fermentation, intracellular ATP level was 1.8 times higher than that of the original strain, while reactive oxygen species (ROS) was reduced by 10%. The destruction of IDP2 gene resulted in rapid autolysis and a decrease in the supply of NADPH. Anti-autolytic index after 96 h autolysis was 4.03 and the NADPH/NADP⁺ ratio was 0.89. After fermentation, intracellular ATP level was reduced by 8% compared with original strain, ROS was 1.3 times higher than that of the original strain. The results may help understand the regulation mechanism of citric acid cycle-related genes on yeast autolysis and provide a basis for the selection of excellent yeast with controllable anti-autolytic performance.
Humans ; Isocitrate Dehydrogenase/genetics* ; NADP ; Reactive Oxygen Species ; Autolysis ; Adenosine Triphosphate

Objective:To observe the effect of escitalopram combined with repetitive transcranial magnetic stimulation (rTMS) on efficacy and attention function in patients with first-episode unipolar depression.Methods:Fifty-two first-episode initial-naive unipolar depression patients were enrolled in Department of Neurology of Guangzhou First People's Hospital from March 2022 to April 2023 were chosen. They were randomly allocated to active stimulation group ( n=27) and sham stimulation group ( n=25); both were treated with escitalopram, and active treatment or sham treatment in the left dorsolateral prefrontal cortex (DLPFC) were given for 4 weeks (5 d per week, 20 d totally). Before treatment and 2 and 4 weeks after treatment, Hamilton Depression Rating Scale (HAMD)-24 was used to evaluate depressive symptoms, and Birmingham Cognitive Screening Scale-Chinese (BCoS-C) was used to evaluate the attention function. Results:(1) In terms of depressive symptoms: HAMD-24 scores of the active stimulation group 2 and 4 weeks after treatment (20.63±2.73, 15.85±2.43) were significantly lower than those before treatment (25.74±2.68, P<0.05); HAMD-24 scores of sham stimulation group 4 weeks after treatment were also significantly lower than those before treatment ([20.48±2.33] vs. [25.80±2.57], P<0.05); HAMD-24 scores of the active stimulation group 2 and 4 weeks after treatment were significantly lower than those of sham stimulation group ( P<0.05). (2) In terms of auditory attention indicators: total correct number (selective attention) in active stimulation group 4 weeks after treatment was significantly larger than that before treatment (51.74±1.38 vs. 47.48±1.60), and the sustained index (sustained attention) was significantly lower than that before treatment (0.74±0.71 vs. 4.37±1.15, P<0.05); total correct number in active stimulation group 4 weeks after treatment was significantly larger than that in sham stimulation group (48.00±1.66), and the sustained index was significantly lower than that in sham stimulation group (3.72±1.28, P<0.05). Conclusion:Combined with escitalopram, rTMS can more effectively mitigate the depressive symptoms in first-episode unipolar depression patients, and depressive symptoms improve more quickly than attentional function.

Objective:To investigate the safety, economic effect, and short-term clinical efficacy of ambulatory total hip arthroplasty (THA) in the treatment of hip osteopathy within.Methods:This study retrospectively reviewed patients who underwent 48-hour outpatient THA and conventional primary THA from July 2020 to July 2021. Gender, age, body mass index (BMI), clinical diagnosis, place of resident, length of hospital stay, duration of the surgery, hemoglobin, albumin, C-reactive protein (CRP) and D-dimer before and 1 day after surgery, the visual analogue scale (VAS) was obtained postoperative day 2, hip joint modified Harris score before and 1 month after surgery, readmission and reoperation within 1 month after operation as the data of evaluations were extracted and compared in this study.Results:A totally of 150 cases were involved in this study, which including 75 cases with 48-hour outpatient primary THA and 75 cases with conventional primary THA. There was no significant difference between the two groups in terms of age, sex, BMI, education level, preoperative diagnosis, and preoperative Harris score ( P>0.05), but a significant difference was found in term of residence (χ 2=6.29, P=0.043), that the patients in the outpatient group were all from Zhejiang Province and 48% (36/75) of them were from Hangzhou City. While, in the conventional group, 6 patients were from other provinces. The length of stay was 2.13±0.52 days and operation time was 59.73±18.91 minutes in the outpatient group, which were both shorter than those (6.71±1.44 days and 66.91±22.40 min) in the conventional group ( t=25.91, P<0.001 for the length of hospital stay; t=2.12, P=0.036 for operation time). Compared with the conventional group, outpatient group saved the average hospital cost (4.60±0.44 vs. 5.20±0.72 ten thousand yuan, t=6.16, P<0.001). The VAS on the second day after surgery 3.45±0.75 was higher in the outpatient group than in the conventional group (3.45±0.75 vs. 3.16±0.94 points, t=2.09, P=0.039). The modified Harris score was without statistical significance ( t=0.42, P=0.677) 1 month after surgery in both groups. 75 patients in the outpatient group, 7 patients delayed discharge (were not discharge within 48 hours), and the rate of delayed discharge was 9.3%. Reasons for delayed discharge included poor pain control in two cases, one case had postoperative nausea and vomiting, one case had failed to meet rehabilitation standards, hypotension in one case, the intraoperative infection in one case and postoperative fever within 48 hours in one case. Conclusion:Outpatient THA can reduce the length of hospital stay, operative time and total cost of hospitalization. It has similar safety and early clinical efficacy as conventional THA. However, a small proportion of patients would delay discharge.

Hemodynamics is a branch of fluid mechanics, which mainly studies the physiological process of circulatory system from the perspective of mechanics, and has a wide range of applications in medicine. The entity teaching model is made to help medical students master the knowledge of hemodynamics more easily and intuitively by showing them the phenomenon of laminar flow, turbulence and axial concentration of red blood cells. And then we check their learning effect by questionnaires. Students are interested in the entity teaching model that the teacher has used in the class. Thus, the entity teaching model can play an important role in the teaching process.

Objective:To explore the correlations of α-melanocyte stimulating hormone ( α-MSH) levels in serum and synovial fluid with progression of primary knee osteoarthritis (KOA). Methods:A retrospective analysis was conducted of the 96 patients who had been diagnosed as primary KOA at Department of Orthopedics, The First Hospital of Huizhou from October 2018 to October 2019. Radiographic severity of KOA was determined by Kellgren-Lawrence (K-L) grades; α-MSH levels were measured by enzyme-linked immunosorbent assay (ELISA). Levels of pro-inflammatory cytokine interleukin-1 β (IL-1 β) and matrix metalloproteinase-3 (MMP-3) were also detected. Another 64 patients with patellar dislocation, matched in age and gender, were enrolled as controls. The Numeric Pain Scale (NPS) and revised Oxford Knee Score (OKS) were employed to evaluate their symptomatic severity. Receiver operating characteristics (ROC) curve was used to compare α-MSH, IL-1 β and MMP-3 with regard to their diagnostic values in the K-L grading. Results:There were no statistically significant difference in age, gender and body mass index between the 2 groups, showing they were comparable ( P> 0.05). The α-MSH levels in synovial fluid were significantly lower in the KOA patients than in the controls [(16.9±3.8) pg/mL versus (18.8±2.7) pg/mL] ( P<0.001); there were no significant differences between the KOA patients and the controls in the serum α-MSH levels [(24.9±1.8) pg/mL versus (24.8±1.7) pg/mL] ( P>0.05). The α-MSH levels in synovial fluid were negatively correlated with K-L grades ( r=-0.382, P<0.001) and negatively correlated with NPS ( r=-0.382, P<0.001) but positively correlated with OKS ( r=0.339, P<0.001). Moreover, the α-MSH levels in synovial fluid were negatively correlated with the IL-1 β levels in synovial fluid ( r=-0.483, P<0.001) and with the MMP-3 levels in synovial fluid ( r=-0.336, P< 0.001). Conclusions:The level of serum α-MSH may not be correlated with the progression of KOA but the synovial fluid α-MSH is negatively correlated with the progression of KOA. Therefore, the expression level of α-MSH in joint synovial fluid can be used as a potential biomarker for assessment of severity of knee osteoarthritis.

With the development of social economy, high-intensity energy is the direct cause of fracture, such as traffic accident, fall injury and tumble. Fracture healing is affected by many factors, such as malunion, delayed union, nonunion and wound infection. Tibial fractures are usually caused by high-energy mechanisms and can also be caused by low-energy mechanisms of ankle rotation. With the gradual deepening of orthopedic surgeons′ understanding of bone nonunion, the treatment of tibial nonunion is mainly divided into surgical treatment and non-operative treatment. Surgical treatment is generally the first choice for the treatment of bone nonunion, in which plate internal fixation plays an unique advantage in the treatment of tibial nonunion. By reviewing the recently published literature, this paper reviews the latest progress in the treatment of tibial nonunion with plate internal fixation.

Objective:To investigate the correlation of mRNA and protein levels of Ghrelin in shoulder tissue and synovial fluid with severity of rotator cuff tear(RCT) and frozen shoulder(FS).Methods:Recruited for this study were 66 RCT and 66 FS patients who had been definitely diagnosed at Department of Orthopedics, The First Hospital of Huizhou from January 2018 to September 2019. Another 60 patients with rotator instability were recruited as a control group. After the severity of RCT was evaluated by ultrasonographic images, the RCT group was further divided into 3 subgroups according to the severity of RCT: a massive full thickness tear (MFTT) subgroup, a non massive full thickness tear (MFTT) subgroup, and a partial thickness tear (PTT) subgroup. The FS group was conventionally divided into 31 cases of acute phase and 35 cases of adhesive phase. Samples of subacromial bursa and shoulder joint bursa tissues and shoulder joint synovial fluid were collected. The expression of Ghrelin mRNA was detected by RT-PCR and the expression of Ghrelin protein in joint synovial fluid was detected by ELISA. The symptoms and functions of shoulder joint were evaluated by visual analogue scale (VAS) and Constant-Murley functional score.Results:There were no significant differences in gender or age between the RCT, FS and control groups, showing compatibility ( P>0.05).The expression of Ghrelin mRNA in subacromial synovial capsule and shoulder joint capsule and the expression of Ghrelin protein in shoulder synovial fluid in the RCT and FS groups were significantly lower than those in the control group ( P<0.05).The expression of Ghrelin mRNA in subacromial synovial capsule and shoulder joint capsule and the expression of Ghrelin protein in shoulder synovial fluid in the PTT subgroup were the highest, followed sequentially by NMFTT and MFTT subgroups, with significant differences between subgroups ( P<0.05). For FS patients, the expression levels of Ghrelin mRNA and protein in the acute phase were significantly lower than in the adhesive phase ( P<0.05). The relative expression of Ghrelin mRNA in joint capsule and the expression of Ghrelin protein in shoulder synovial fluid were negatively correlated with VAS scores and IL-6 levels (all P<0.05), and positively correlated with the Constant-Murley scores (all P<0.05). Conclusions:The expression of Ghrelin in shoulder tissue and synovial fluid is negatively correlated with the progress of rotator cuff tear and frozen shoulder.Local supplementation of Ghrelin may be a potential therapy for rotator cuff tear and frozen shoulder.