ObjectiveTo explore the effect of Shenge Bushen Capsules （SBC） on sexual development in normal 3-week-old mice. MethodsThe experiment consisted of two parts. In the first part， mice were divided into four groups： The control group and the low， medium， and high-dose SBC groups （234.7， 469.4， 938.7 mg·kg^-1， respectively）. In the second part， mice were divided into four groups： Control group， Pseudostellariae Radix polysaccharide （PRP） group， total flavonoids group， and SBC group， all receiving a dose of 469.4 mg·kg^-1. After 7 days of administration， the vaginal opening of female mice and the descent of testes and scrotum in male mice， as well as the ovarian and testicular organ indices， were observed. After 4 weeks of administration， female and male mice were housed together for 2 days， and the pregnancy rate of females was monitored. After delivery， the pregnant female mice continued receiving the treatment for 4 weeks， and the sexual development of their offspring， including vaginal opening， testicular descent， and organ indices of ovaries and testes， was observed. Serum sex hormones were measured by enzyme-linked immunosorbent assay （ELISA）， and the expression of gonadotropin-releasing hormone （GnRH） and growth hormone （GH） proteins in the hypothalamus was assessed by Western blot. ResultsCompared with the control group， there was no significant effect on the vaginal opening of female mice or the descent of testes in male mice after 7 days of SBC administration. After 4 weeks of administration， the pregnancy rate in the low-dose group was significantly reduced （P<0.05）， but no significant effects were observed in the other groups. The three doses of SBC did not significantly affect the ovarian or testicular organ indices， and there was no significant upregulation in the expression of GnRH or GH in the hypothalamus. The primary component of SBC， Pseudostellariae Radix polysaccharide， significantly reduced the vaginal opening in female mice after 7 days of administration （P<0.05）. After 4 weeks， the serum estradiol levels of non-pregnant female mice were decreased （P<0.05）， but there was no significant effect on the expression of GnRH or GH proteins in the hypothalamus of either male or female mice. Additionally， there were no significant effects on precocious puberty indicators， such as vaginal opening and testicular descent， in the offspring mice. ConclusionSBC does not significantly promote precocious puberty in young mice， and it does not have any noticeable effects on the pregnancy rate of adult mice or the sexual development of their offspring.

Metabolic-associated fatty liver disease (MAFLD) and osteoporosis (OP) are two very common metabolic diseases. A growing body of experimental evidence supports a pathophysiological link between MAFLD and OP. MAFLD is often associated with the development of OP. Rutaecarpine (RUT) is one of the main active components of Chinese medicine Euodiae Fructus. Our previous studies have demonstrated that RUT has lipid-lowering, anti-inflammatory and anti-atherosclerotic effects, and can improve the OP of rats. However, whether RUT can improve both fatty liver and OP symptoms of MAFLD mice at the same time remains to be investigated. In this study, we used C57BL/6 mice fed a high-fat diet (HFD) for 4 months to construct a MAFLD model, and gave the mice a low dose (5 mg·kg^-1) and a high dose (15 mg·kg^-1) of RUT by gavage for 4 weeks. The effects of RUT on liver steatosis and bone metabolism were then evaluated at the end of the experiment [this experiment was approved by the Experimental Animal Ethics Committee of Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences (approval number: IMB-20190124D₃03)]. The results showed that RUT treatment significantly reduced hepatic steatosis and lipid accumulation, and significantly reduced bone loss and promoted bone formation. In summary, this study shows that RUT has an effect of improving fatty liver and OP in MAFLD mice.

Objective To summarize the experience and practical value of living donor kidney harvesting in Bama miniature pigs with six gene modified. Methods The left kidney of Bama miniature pigs with six gene modified was obtained by living donor kidney harvesting technique. First, the ureter was occluded, and then the inferior vena cava and abdominal aorta were freed. During the harvesting process, the ureter, renal vein and renal artery were exposed and freed in sequence. The vascular forceps were used at the abdominal aorta and inferior vena cava, and the renal artery and vein were immediately perfused with 4℃ renal preservation solution, and stored in ice normal saline for subsequent transplantation. Simultaneously, the donor abdominal aorta and inferior vena cava gap were sutured. The operation time, blood loss, warm and cold ischemia time, postoperative complications and the survival of donors and recipients were recorded. Results The left kidney of the genetically modified pig was successfully harvested. Intraoperative bleeding was 5 mL, warm ischemia time was 45 s, and cold ischemia time was 2.5 h. Neither donor nor recipient pig received blood transfusion, and urinary function of the kidney transplanted into the recipient was recovered. The donor survived for more than 8 months after the left kidney was resected. Conclusions Living donor kidney harvesting is safe and reliable in genetically modified pigs. Branch blood vessels could be processed during kidney harvesting, which shortens the process of kidney repair and the time of cold ischemia. Living donor kidney harvesting contributes to subsequent survival of donors and other scientific researches.

Objective:To investigate the safety and efficacy of pseudomonas aeruginosa Injection (PAI) in the treatment of coeliac leakage after cervical lymph node dissection for robotic thyroid cancer.Methods:Retrospective analysis of 1262 patients who underwent robotic thyroid surgery at the 960th Hospital of the People’s Liberation Army from Jan. 2021 to Aug. 2023 was made. Postoperative celiac leakage happened in 28 patients. The control and injection groups were divided according to whether pseudomonas aeruginosa injection was used or not. In the control group, there were 4 males and 11 females out of 15 patients, with an average age of (46.20±9.02) years old, and the treatment methods of low-fat diet, negative pressure suction, and pressure bandage were used. In the injection group, there were 7 males and 6 females among 13 patients, with an average age of (41.00±8.87) years. They were treated with low-fat diet, negative pressure suction, pressure bandage, and PAI .The number of lymph node dissection, total drainage volume, peak drainage volume, days of hospitalization and the rate of decline in drainage volume within 24 h after the use of PAI in the injection group, post-injection temperature, number of injections, and post-injection extubation time were statistically analyzed in both groups. Independent samples t test, Mann-Whitney U test, χ2 test or Fisher’s exact test were used for comparison between groups. Results:There was no statistically significant difference in age, gender, extent of thyroidectomy, highest daily drainage volume, and total drainage volume between the two groups ( P>0.05). The difference in the number of lymph node dissection (49.15±23.05 vs. 30.80±11.76, P=0.012) and hospitalization time (11.77±4.64 vs. 16.40±6.42, P=0.041) between the injection group and the control group was statistically significant. After the use of pseudomonas aeruginosa injection, the draining fluid decreased from the previous day (69.56± 20.82) % in the injection group; Twelve patients were successfully extubated after one injection of pseudomonas aeruginosa injection, and one patient was successfully extubated after two injections; the mean time to extubation after injection was (3.85±1.28) days; 76.9% patients (10/13) had fever symptoms within 48 h with body temperature (38.05±0.89) ℃ after injection, body temperature returned to normal after symptomatic treatment,and no other adverse reactions occurred except fever. Conclusion:Pseudomonas aeruginosa injection is safe and effective in the treatment of celiac leakage after cervical lymph node dissection for robotic thyroid cancer, effectively reducing cervical drainage and shortening hospitalization days.

ObjectiveTo observe the effect of acupuncture “Zhibian （BL 54）-to-Shuidao （ST 28）” on the reproductive function of asthenospermia model mice and to explore the possible mechanism. MethodsForty-two male C57BL/6 mice were randomly divided into blank group， model group， and acupuncture group， with 14 mice in each group. Cyclophosphamide 30 mg/（kg·d） was injected intraperitoneally for 7 days to establish model of asthenospermia for the model group and the acupuncture group， while 0.9% sodium chloride solution 10 ml/（kg·d） was injected intraperitoneally for 7 days in the blank group. After successful modeling， mice in the acupuncture group received acupuncture at “Zhibian （BL 54）-to-Shuidao （ST 28）” once a day for 20 minutes， 6 times a week for 3 weeks； the remaining two groups were fixed without acupuncture. Daily observations were made on the general conditions of mice in all groups， and changes in body weight after intervention were recorded. On the next day after completing the treatment， 6 male mice selected randomly from each group to test the sperm quality as well as testicular and epididymal weights， and calculate the corresponding indices； ELISA was used to test the levels of testosterone （T）， follicle-stimulating hormone （FSH）， and luteinizing hormone （LH） in serum； HE staining and TUNEL staining were performed to observe the pathological morphology and apoptosis of testicular and epididymal tissues； fluorescence quantitative PCR and western blot were used to detect changes in the expression of apoptosis-related factors （Fas）， Fas-associated death domain protein （FADD）， B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated X protein （Bax）， and cysteine-aspartic acid protease （caspase）-3 mRNA and protein in testicular tissue. The remaining 8 male mice in each group were housed with estrus-cycling mice of the same strain at 1∶1 ratio， and the pregnancy rate and number of embryos per litter in each group were determined after mating. ResultsIn comparison with the blank group， mice in the model group exhibited reduced body weight， decreased testicular mass， testicular index， epididymal mass， and epididymal index. Additionally， there was a decrease in total sperm count， sperm motility， and sperm viability. Serum levels of T， FSH， and LH were reduced. The apoptosis rate increased， and the expression levels of Fas， FADD， Bax， and Caspase-3 mRNA and proteins were elevated， while Bcl-2 mRNA and protein expression levels decreased （P＜0.01）. Pathological abnormalities were observed in testicular and epididymal tissues， with disrupted arrangement of seminiferous tubules and a decreased number of spermatogenic cells within the tubular lumen. Furthermore， the pregnancy rate and the number of embryos per litter decreased significantly after mating （P＜0.01）. In comparison with the model group， mice in the acupuncture group showed improvements in testicular mass， testicular index， epididymal mass， epididymal index， total sperm count， sperm motility， and sperm viability. Serum levels of T， FSH， and LH increased. The apoptosis rate decreased， and the expression levels of Fas， FADD， Bax， and Caspase-3 mRNA and proteins decreased， while Bcl-2 mRNA and protein expression levels increased （P＜0.05 or P＜0.01）. Morphological improvements were observed in testicular and epididymal tissues， with a compact arrangement of seminiferous tubules and an increased number of spermatogenic cells within the tubular lumen. Furthermore， the pregnancy rate and the number of embryos per litter increased significantly after mating （P＜0.01）. ConclusionAcupuncture “Zhibian （BL 54）-to-Shuidao （ST 28）” can improve testicular tissue apoptosis and enhance reproductive function in a mouse model of asthenospermia. Its mechanism may be associated with the modulation of key factors in the extrinsic membrane receptor pathway （Fas-mediated pathway） and the intrinsic mitochondrial pathway （Bcl-2/Bax-regulated pathway） in testicular tissue.

convalescents, and to screen for broad-spectrum neutralizing antibodies against the SARS-CoV-2 RBD. Methods Using biotinylated RBD as a molecular probe, flow cytometry was employed to perform single-cell sorting of B cells from peripheral blood mononuclear cells (PBMCs) of convalescents. The obtained B cells were lysed and subjected to reverse transcription, followed by nested PCR amplification of the heavy and light chains of antibodies was conducted using random primers. The amplified products were cloned into corresponding expression vectors, and the respective matched heavy-light chain plasmids were co-transfected into 293F cells for expression. Monoclonal antibodies were then purified using Protein A column chromatography. Neutralization experiments were conducted with the wild-type (WT) pseudovirus, and antibodies with IC50<0.1 μg/mL were selected for further testing of neutralizing breadth and potency against the wild-type (WT), Beta variant (B.1.351), Delta variant (B.1.617.2), and currently prevalent pseudovirus strains (XBB, BA.5, BF.7). Results A total of 21 RBD-specific monoclonal B cells were obtained from two recovered patients, resulting in the isolation of 13 pairs of antibody light/heavy chains. Nine antibodies were successfully expressed, with P1-A1, P1-B6, and P1-B9 exhibiting IC50 values below 0.1 μg/mL against the pseudovirus of the wild-type strain (WT). Specifically, P1-B6 effectively neutralized the wild-type strain (WT), Beta variant (B.1.351), and Delta variant (B.1.617.2), with IC50 values reaching 0.01 μg/mL. P1-B9 demonstrated effective neutralization against the wild-type strain (WT), Beta variant (B.1.351), Delta variant (B.1.617.2), and Gamma variant (P.1) pseudoviruses, with IC50 values of 0.42 μg/mL, 0.63 μg/mL, 0.28 μg/mL, and 2.50 μg/mL, respectively. Additionally, P1-B6 exhibited good neutralization against BA.5 and BF.7 pseudoviruses, with IC50 values of 0.06 μg/mL and 0.09 μg/mL, respectively. Conclusions Infection with the SARS-CoV-2 WT strain can induce the generation of neutralizing antibodies with broad-spectrum activity. Generating these broadly neutralizing antibodies does not require an excessively high somatic hypermutation. The obtained antibodies can be used as candidates for SARS-CoV-2 diagnosis and prevention.

Objective To compare the efficacy of TightRope loop plate and Endobutton plate in the treatment of acromioclavicular joint dislocation.Methods A retrospective analysis was conducted on 94 patients with acromioclavicular joint dislocation who were treated at this center from March 2021 to February 2023.They were divided into two groups based on different admission date.The Group E(n =47)received Endobutton plate treatment between March 2021 and February 2022,while the Group T(n =47)received TightRope loop plate treatment between March 2022 and February 2023.At the last follow-up,the perioperative indicators,Visual Analogue Scale(VAS),Constant-Murley shoulder joint function scores,and surgical complications were compared between the two groups.Results The surgical time,intraoperative bleeding,incision length,and VAS scores at 7 days after surgery in the Group T were shorter or lower than those in the Group E(P<0.05).There were no statistical differences in the incidence of perioperative nerve injury,internal fixation displacement,clavicle fracture,vascular injury,and infection between the two groups(P>0.05).The subjective and objective scores of Constant-Murley shoulder joint function in both groups at9 months after surgery showed significant improvement compared to preoperative scores(all P =0.000).There was no significant difference in the subjective and objective scores of Constant-Murley shoulder joint function between the two groups at 9 months after surgery(P>0.05).Conclusions The treatment of acromioclavicular joint dislocation with TightRope loop plate ot or Endobutton plate has a significant effect and can effectively improve shoulder joint function.Compared with Endobutton plate,use of TightRope loop plate has minor surgical trauma,less bleeding,and significantly reduced postoperative pain,being more conducive to early functional exercise for patients.

With the increasing proportion of human papilloma virus(HPV)infection in the pathogenic factors of oro-pharyngeal cancer,a series of changes have occurred in the surgical treatment.While the treatment mode has been im-proved,there are still many problems,including the inconsistency between diagnosis and treatment modes,the lack of popularization of reconstruction technology,the imperfect post-treatment rehabilitation system,and the lack of effective preventive measures.Especially in terms of treatment mode for early oropharyngeal cancer,there is no unified conclu-sion whether it is surgery alone or radiotherapy alone,and whether robotic minimally invasive surgery has better func-tional protection than radiotherapy.For advanced oropharyngeal cancer,there is greater controversy over the treatment mode.It is still unclear whether to adopt a non-surgical treatment mode of synchronous chemoradiotherapy or induction chemotherapy combined with synchronous chemoradiotherapy,or a treatment mode of surgery combined with postopera-tive chemoradiotherapy.In order to standardize the surgical treatment of oropharyngeal cancer in China and clarify the indications for surgical treatment of oropharyngeal cancer,this expert consensus,based on the characteristics and treat-ment status of oropharyngeal cancer in China and combined with the international latest theories and practices,forms consensus opinions in multiple aspects of preoperative evaluation,surgical indication determination,primary tumor re-section,neck lymph node dissection,postoperative defect repair,postoperative complication management prognosis and follow-up of oropharyngeal cancer patients.The key points include:① Before the treatment of oropharyngeal cancer,the expression of P16 protein should be detected to clarify HPV status;② Perform enhanced magnetic resonance imaging of the maxillofacial region before surgery to evaluate the invasion of oropharyngeal cancer and guide precise surgical resec-tion of oropharyngeal cancer.Evaluating mouth opening and airway status is crucial for surgical approach decisions and postoperative risk prediction;③ For oropharyngeal cancer patients who have to undergo major surgery and cannot eat for one to two months,it is recommended to undergo percutaneous endoscopic gastrostomy before surgery to effectively improve their nutritional intake during treatment;④ Early-stage oropharyngeal cancer patients may opt for either sur-gery alone or radiation therapy alone.For intermediate and advanced stages,HPV-related oropharyngeal cancer general-ly prioritizes radiation therapy,with concurrent chemotherapy considered based on tumor staging.Surgical treatment is recommended as the first choice for HPV unrelated oropharyngeal squamous cell carcinoma(including primary and re-current)and recurrent HPV related oropharyngeal squamous cell carcinoma after radiotherapy and chemotherapy;⑤ For primary exogenous T1-2 oropharyngeal cancer,direct surgery through the oral approach or da Vinci robotic sur-gery is preferred.For T3-4 patients with advanced oropharyngeal cancer,it is recommended to use temporary mandibu-lectomy approach and lateral pharyngotomy approach for surgery as appropriate;⑥ For cT1-2N0 oropharyngeal cancer patients with tumor invasion depth＞3 mm and cT3-4N0 HPV unrelated oropharyngeal cancer patients,selective neck dissection of levels ⅠB to Ⅳ is recommended.For cN+HPV unrelated oropharyngeal cancer patients,therapeutic neck dissection in regions Ⅰ-Ⅴ is advised;⑦ If PET-CT scan at 12 or more weeks after completion of radiation shows intense FDG uptake in any node,or imaging suggests continuous enlargement of lymph nodes,the patient should undergo neck dissection;⑧ For patients with suspected extracapsular invasion preoperatively,lymph node dissection should include removal of surrounding muscle and adipose connective tissue;⑨ The reconstruction of oropharyngeal cancer defects should follow the principle of reconstruction steps,with priority given to adjacent flaps,followed by distal pedicled flaps,and finally free flaps.The anterolateral thigh flap with abundant tissue can be used as the preferred flap for large-scale postoperative defects.

Objective:To explore the expression pattern of pyroptosis-related genes(PRGs)in hepatocellular carcinoma(HCC),and analyze the relationship between its expression and tumor prog-nosis and immune microenvironment.Methods:TCGA database was used to analyze the genetic changes and expression patterns of PRGs in primary HCC cells,and cluster analysis was used to i-dentify the pyrogenic subtypes of HCC.To compare the difference of prognosis and immune mi-croenvironment among HCC pyrodeath subtypes.Scorch death score quantified the comprehensive expression of PRGs in each sample,and analyzed the correlation between scorch death score and each immune score.Results:Two pyroptosis-associated subtypes of primary HCC were identi-fied,and the expression pattern of PRGs is closely related to the prognosis of cancer patients and the tumor microenvironment.The subtype with high expression of PRGs had a poor prognosis,and functional enrichment analysis found that some tumor-promoting pathways and PD-1 checkpoint pathways were significantly enriched in this subtype.And various cells and immune checkpoints re-lated to immunosuppression were also enriched in this subtype.By constructing PYROPTO-SIS_score to quantify the comprehensive expression of pyroptosis-related genes in each sample,it was found that PYROPTOSIS_score was significantly positively correlated with tumor-infiltrating macrophages,myeloid-derived suppressor cells,and Treg cells.Conclusion:These results sug-gest that pyroptosis may play a tumor-promoting as well as immunosuppressive role in HCC,pro-viding new insights into the assessment of tumor patient prognosis and the immune microenviron-ment.

Objective:To explore the correlation between high cholinergic pathway signaling and cognitive function in patients with Parkinson disease(PD) accompanied with sleep disorder.Methods:PD patients admitted from 2017 to 2022 were divided into PD with sleep disorder group (PD-SD group) ( n=56) and PD without sleep disorder group (PD-NSD group) ( n=41) according to the Parkinson's disease sleep scale (PDSS) score. All participants underwent magnetic resonance imaging examination.All patients were evaluated by the PDSS, Hoehn-Yahr (H-Y), Montreal cognitive assessment scale (MoCA), and cholinergic pathways hyper intensities scale (CHIPS). The difference of cognitive function between the two groups and the correlation between CHIPS and cognitive function were analyzed.Independent sample t-test, Spearman correlation analysis, and binary Logistic regression analysis were performed on the data by SPSS 26.0 statistical software. Results:(1)The MoCA score of the PD-SD group (22.00 (5.00)) was lower than that of the PD-NSD group (26.00 (5.00)) ( Z=-3.830, P<0.05). The total and all aspects scores of CHIPS in PD-SD group were higher than those in PD-NSD group(the total score of the low external capsule: 12.00(8.00), 0(8.00), the total score of the high external capsule: 12.00(2.00), 6.00(9.00), the total score of the radial crown: 8.00(0), 4.00(4.00), the total score of the centrum semiovale: 3.00(4.00), 0(2.00), the total score of the right side: 16.00(9.00), 5.00(10.00), the total score of the left side: 17.00(6.00), 7.00(9.00), the total score of CHIPS: 32.00(14.00), 14.00(20.00))( Z=-5.081, -5.873, -4.933, -3.211, -5.562, -6.232, -5.995, all P<0.05). (2)The correlation analysis between the score of CHIPS and cognitive function in the PD-SD group showed that, the total score of the low external capsule ( r=-0.286), the total score of the centrum semiovale ( r=-0.307), the total score of the right side ( r=-0.376), the total score of the left side ( r=-0.284) and the total score of CHIPS ( r=-0.349) were negatively correlated with MoCA(all P<0.05). (3)Binary Logistic regression analysis showed that white matter lesions in centrum semiovale, low inner capsule, right and left leukodystrophy were not influence factors for cognitive impairment (all P>0.05). Conclusion:PD patients with sleep disorders have lower cognitive function scores, higher CHIPS scores, and significant changes in white matter lesions compared to those without sleep disorders. In PD patients with sleep disorders, the higher the CHIPS score, the lower the cognitive function score, and the more significant the rate of cognitive impairment occurrence and development.