Liver transplantation (LT) has become a standard treatment for end-stage liver diseases, and graft injury is intricately associated with poor prognosis. Granzyme B (GZMB) plays a vital role in natural killer (NK) cell biology, but whether NK-derived GZMB affects graft injury remains elusive. Through the analysis of single-cell RNA-sequencing data obtained from human LT grafts and the isolation of lymphocytes from mouse livers following ischemia-reperfusion injury (IRI), we demonstrated that 2NK cells with high expression of GZMB are enriched in patients and mice. Both systemically and liver-targeted depletion of NK cells led to a notable reduction in GZMB⁺ cell infiltration, subsequently resulting in diminished graft injury. Notably, the reconstitution of Il2rg ^-/- Rag2 ^-/- mice with purified Gzmb-KO NK cells demonstrated superior outcomes compared to those with wild-type NK cells. Crucially, global knockout of GZMB and pharmacological inhibition exhibited remarkable improvements in liver function in both mouse IRI and rat LT models. Moreover, a phosphorylated derivative of FDA-approved vidarabine was identified as an effective inhibitor of mouse GZMB activity by molecular dynamics, which could provide a potential avenue for therapeutic intervention. Therefore, targeting NK cell-derived GZMB during the LT process suggests potential therapeutic strategies to improve post-transplant outcomes.

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 evaluate the validity and reliability of the Chinese version of the Lubeck Alcohol Withdrawal Risk Scale(C-LARS).Methods:Referring to previous literature and clinical expert opinions using the Delphi method,a C-LARS was established through translation and back-translation.Principal component analysis was used to evaluate the structural validity of C-LARS,and the maximum variance method was used to calculate factors and factor loadings to evaluate the structural validity of C-LARS.The severity of alcohol withdrawal syn-drome in patients was evaluated using the the Alcohol Withdrawal Scale(AWS),and the criterion validity of C-LARS was evaluated by calculating the correlation coefficient between AWS and C-LARS.Results:The internal correlation coefficient(ICC)of the evaluation factors was 0.972,and the total Cronbach α coefficient was 0.938.Factor analysis and ROC analysis showed that a C-LARS score of ≥ 3 could predict the occurrence of mild alcohol withdrawal syndrome,with sensitivity and specificity of 0.945 and 0.899,respectively.The C-LARS scores of ≥ 5 could predict the occurrence of moderate alcohol withdrawal syndrome,with sensitivity and specificity of 0.910 and 0.905,respectively.The C-LARS scores of≥7 could predict the occurrence of severe alcohol with-drawal syndrome within 12 hours,the sensitivity and specificity were 0.990 and 0.877,respectively.Conclusion:The C-LARS has good validity and reliability,which could be used as a tool to assess the incidence and severity of alcohol withdrawal syndrome within 12 hours after visit.

Objective To evaluate the diffusion distribution of bone cement in the vertebral body by quadrant method,and to analyze and evaluate the correlation between the diffusion distribution type of bone cement and new vertebral fractures after vertebral augmentation.Methods A total of 170 subjects who met the conditions from January 2020 to December 2021 were collected.According to the anteroposterior and lateral view of the spine,the injured vertebra was divided into four quadrants,and divided into homogeneous diffusion group and uneven diffusion group according to the postoperative diffusion distribution of bone cement in the injured vertebra.The incidence and types of refracture were followed up,and the VAS score and Cobb angle were compared between the two groups.Results 170 patients were followed up for at least 12 months,including 90 patients in homogeneous diffusion group and 80 patients in heterogeneous diffusion group.There were 33 cases of refracture(19.41％),12 cases of refracture(13.33％)in the diffuse homogeneous group,and 21 cases of refracture(26.25％)in the diffuse heterogeneous group,and the difference between the groups was statistically significant(P＜0.05).The site of refracture in the diffuse homogeneous group was mainly the clinical vertebral fracture,while the probability of refracture in the diffuse heterogeneous clinical vertebra and the operated vertebra was similar.The incidence of postoperative bone cement leakage in the diffuse homogeneous group was significantly lower than that in diffuse heterogeneous group(P＜0.05).The VAS score and Cobb angle were significantly improved in both groups after surgery and at the last follow-up compared with those before surgery,but there was no significant difference between groups.Conclusion The incidence of new vertebral fractures after vertebroplasty is closely related to the type of cement diffusion,and the risk of refracture defined as uneven cement diffusion by quadrant method is high.

The clinical data of a child with MORC2 gene mutation related neurodevelopmental disorder treated in Fujian Medical University Union Hospital in July 2020 were analyzed retrospectively.The male (7-year-old)patient was global retardation from infant, with special face, short stature, small head circumference, decreased muscle strength and positive pyramidal tract sign of lower limbs.Brain magnetic resonance imaging was similar to the changes of Leigh syndrome.Genetic testing found de novo mutation in MORC2 gene chr22: 31345763, c.292G>A(p.Gly98Arg). And literature review found that there was only one related report. MORC2 gene mutation related neurodevelopmental disorder is a newly discovered syndrome, and c. 79G>A(p.Glu27Lys) is the most common mutation.This case enriched the clinical phenotype and genotype of neurodevelopmental disorder related to MORC2 gene.

Objective:To investigate the prognostic significance of tumor architecture in patients with upper tract urothelial carcinoma (UTUC) after radical nephroureterectomy.Methods:A retrospective study was performed on 958 patients who underwent nephroureterectomy in Second Affiliated Hospital of Zhejiang university (156) and Renji Hospital (802) between January 1998 and June 2019. There were 630 males and 328 females with median age 67 years old, ranging 30-89 years old. Among them, 499 patients suffered with preoperative hydronephrosis, 370 patients suffered with hypertension, 120 patients suffered with diabetes, 252 patients had history of smoking and 119 patients had history of non-muscle invasive bladder cancer (NMIBC) or with NMIBC. 489 patients had tumor in renal pelvic, 394 patients had tumor in ureter and 75 patients had tumor in both sites. Laparoscopic surgery was performed in 543 patients while open surgery was performed in 415 patients. The χ 2 test was used to detect the association between tumor architecture and several clinicopathological features. Kaplan-Meier method with the log-rank test was used to assess survival analysis. Multivariate analyses were conducted using Cox proportional-hazards regression model. Results:516 cases (53.9%) showed papillary architecture(Group A) and 442 cases (46.1%) showed sessile architecture(Group B). 543 patients had a tumor ≤3 cm and 415 had a tumor >3 cm. Low pathological grade and high grade was diagnosed in 275 and 683 patients, respectively. The distribution of pathological stage was pT a-1 in 441 cases, pT 2 in 180 cases, pT 3 in 308 cases and pT 4 in 29 cases. Lymphadenectomy was performed in 227 patients and 62 patients were pathologically confirmed lymph node metastasis. 48 patients were found squamous or glandular differentiation. Lymphovascular invasion (LVI) was observed in 150 patients. 134 patients were multifocality. Positive surgical margin was found in 43 patients. Median follow-up was 39 (ranging, 2-206) months. During follow-up, a total of 304 patients died and 236 died of UTUC. 5-year OS and CSS were 76.6% and 81.8%, respectively, in patients with papillary architecture (group A), which were significantly higher than 54.4% and 60.5% in patients with sessile architecture (group B, all P<0.001). Patients in group B had more female patients (38.9% vs.30.3%, P=0.005), ureteral location (47.1% vs. 36.1, P=0.002), hydronephrosis (55.9% vs.48.8%, P=0.030) and postoperative adjuvant chemotherapy (27.1% vs. 14.7%, P<0.001), higher pathological grade (89.6% vs.55.6%, P<0.001) and stage (79.4% vs.32.4%, P<0.001), lymph node metastasis rate (12.0% vs.1.7%, P<0.001), squamous or glandular differentiation (9.5% vs.1.2%, P<0.001) and LVI (24.4% vs.8.1%, P<0.001) than patients in group A. Cox multivariate regression analysis showed that sessile architecture ( P=0.022, 0.028), age ≥65 years ( P<0.001, <0.001), history of diabetes ( P=0.008, 0.043), history of NMIBC or with NMIBC ( P<0.001, <0.001), higher grade ( P=0.002, <0.001), advanced tumor stage ( P=0.003, 0.005), lymph node metastasis ( P=0.003, 0.044), squamous or glandular differentiation ( P=0.008, 0.027) and positive surgical margin ( P=0.003, 0.010) were independent risk factors for OS and CSS. However, tumor >3 cm ( P=0.013, 0.131) and positive LVI ( P=0.045, 0.174) were independent risk factors for CSS rather than OS. Conclusions:UTUC is high malignancy. Tumor architecture was one of an independent risk factor for OS and CSS in UTUC patients and sessile tumors were more malignant, more aggressive and have worse prognosis.

OBJECTIVE: To observe the effect of occlusal interference on the afferent pathway of the trigeminal nerve and neuronal excitability in the trigeminal subnucleus caudalis (SPVC) of rats by electrical stimulation of the trigeminal ganglion (TG) and extracellular recordings of SPVC activities. METHODS: Twenty male Wistar rats were randomly divided into control group and model group (=10). In the model group, occlusal interference for 30 consecutive days was induced using light-cured flowable resin on the right maxillary molars. During occlusal interference, the pain sensitivity was scored with von Frey Fibers in the masseter. Simultaneous recordings of electrical activities from the SPVC, electrocardiogram, body temperature and electromyogram of the breath muscles of the anesthetized rats were performed, and the responses evoked by electrical stimulation of the TG were analyzed. RESULTS: Compared with the control rats, the rats in the model group showed significantly increased pain sensitivity scores ( < 0.05) and increased spontaneous discharge frequency of the SPVC ( < 0.05). The amplitude of the SPVC responses induced by electrical stimulation of the TG showed stimulus intensity-dependent changes ( < 0.05), and the amplitude evoked by 4 mA and 8 mA stimulation was similar between the model group and the control group (>0.05). Train stimulation (0.2 ms, 1 mA, 30 s, 100 Hz) of the TG significantly increased the discharge frequency of the SPVC only in the rats in the model group ( < 0.05). CONCLUSIONS The functional activities of the pain afferent pathway of the trigeminal nerve can be electrophysiologically monitored by electrical stimulation of the TG and extracellular recordings of SPVC activities in rats. Occlusal interference can increase the excitability of the neurons in the SPVC and enhance their sensitivities to TG afferent activation, suggesting the neural plasticity of the pain afferent pathway.

From April 2017 to April 2018, three male patients aged 46-71 years with large area burns were treated in our hospital. Acute acalculous cholecystitis (AAC) symptoms of the patients began to appear 15-81 days after injury. AAC was diagnosed 24-81 days after injury. Ultrasound-guided percutaneous transhepatic cholecystostomy was performed 26-82 days after injury. The symptoms subsided in 2 patients, and cholecystectomy was performed in 1 patient with gallbladder perforation 94 days after injury. The patients were cured and discharged 41-118 days after injury. No recurrence of cholecystitis occurred during 8-9 months of follow-up after discharge.

OBJECTIVE:To establish the method for simultaneous determination of berberine hydrochloride and baicalin in Jianpi zhixiening granules. METHODS:HPLC switching walvelength method was adopted. The determination was performed on Hypersil BDS C18 column with mobile phase consisted of methanol-0.45％ phosphoric acid solution-triethylamine(50:49:1,V/V/V) at the flow rate of 1.0 mL/min. The detection wavelength was set at 265 nm(berberine hydrochloride)and 280 nm(baicalin). The column temperature was set at 30 ℃,and sample size was 10 μL. RESULTS:The linear range of berberine hydrochloride and baicalin were 60.3-312.8 ng(r=0.9997)and 81.5-368.9 ng(r=0.9999). The limits of quantitation were 0.6668,0.7740 ng,andthe limits of detection were 0.2226,0.2580 ng,respectively. RSDs of intermediate precision,stability and repeatability tests were all lower than 1.0％. The recoveries were 96.48％-99.30％(RSD=1.06％,n=6) and 95.20％-99.39％(RSD=1.66％,n=6), respectively. RSDs of durability test were all lower than 2.0％. CONCLUSIONS:The method is simple, precise, stable, reproducible,accurate and durable. It can be used for simultaneous determination of berberine hydrochloride and baicalin in Jianpi zhixiening granules.

OBJECTIVE:To establish the method for simultaneous determination of the content of berberine hydrochloride, baicalin and osthole in Jinchan zhiyang capsules. METHODS:HPLC method was adopted. The determination was performed in Hypersil BDS C18 column with mobile phase consisted of methanol-acetonitrile-0.1%phosphoric acid-three triethylamine(50∶30∶19∶1, V/V/V/V) at the flow rate of 1.0 mL/min. The detection wavelengths were set at 265 nm (berberine hydrochloride),280 nm (baicalin)and 322 nm(osthole). The column temperature was set at 30 ℃,and sample size was 10 μL. RESULTS:The linear range of berberine hydrochloride,baicalin and osthole were 80-800 ng(r＝0.999 8),60-600 ng(r＝0.999 9),60-600 ng(r＝0.999 6),respectively. RSDs of precision,stability and reproducibility tests were all lower than 2.0%. The limits of quantitation were 80,60,60 ng,respectively,and the limits of detection were 24,20,20 ng,respectively. The recovery rates were 97.4%-98.3%(RSD＝0.33%,n＝6),98.4%-99.6%(RSD＝0.42%,n＝6)and 96.9%-99.0%(RSD＝0.92%,n＝6),respectively. RSDs of durability tests were all lower than 1.2%. CONCLUSIONS:The method is simple, accurate, precise, stable, reproducible and durable. It can be used for simultaneous determination of berberine hydrochloride,baicalin and osthole in Jinchan zhiyang capsules.