ObjectiveTo investigate the key targets of Jianpi Yiqi prescription （JYP） in the treatment of hepatocellular carcinoma （HCC） based on network pharmacology and explore the effect of JYP on the invasion and proliferation of hepatocellular carcinoma cells via protein tyrosine phosphatase， non-receptor type 1 （PTPN1） by bioinformatics analysis and CRISPR/Cas9. MethodsThe potential targets of JYP in the treatment of HCC were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）， SwissTargetPrediction， GeneCards， NCBI， and CTD. Additionally， the active components of JYP that could interact with PTPN1 were screened out， and then molecular docking between the targets and active components was performed in Autodock 4.0. UALCAN， HPA， and LinkedOmics were used to analyze the expression of PTPN1 in the HCC tissue， and the relationship of PTPN1 expression with the overall survival （OS） of HCC patients was discussed. CRISPR/Cas9 was used to knock down the expression of PTPN1 in HepG2 and SK-hep-1 cells， and the knockdown effect was examined by sequencing， Real-time PCR， and Western blot. HepG2 cells were classified into blank control， low-， medium-， and high-dose JYP （5.25， 10.5， 21 g·kg^-1）， and PTPN1 knockout groups. Real-time PCR and Western blot were employed to determine the mRNA and protein levels， respectively， of PTPN1 in HepG2 cells of each group. The effects of JYP and PTPN1 knockdown on the proliferation， invasion， and apoptosis of HepG2 cells were detected by Cell Counting Kit-8 （CCK-8）， Transwell， and Annexin V-FITC/PI methods， respectively. ResultsJYP had the most active components targeting PTPN1， and 31 of the active components had the binding energy less than -5.0 kcal·mol^-1 in molecular docking. The mRNA and protein levels of PTPN1 in the HCC tissue were higher than those in the normal tissue （P<0.01）. Compared with that in the normal tissue， the mRNA level of PTPN1 in the HCC tissue was up-regulated at the pathological stages Ⅰ-Ⅲ and grades G₁-G₃ （P<0.01）， and it was not significantly up-regulated at the stage Ⅳ or grade G₄. The mRNA level of PTPN1 in the TP53-mutated HCC tissue was higher than that in the TP53-unmutated HCC tissue （P<0.01）. The high mRNA level of PTPN1 was associated with the OS reduction （P<0.01）. After treatment with the JYP-containing serum or knockdown of PTPN1， HepG2 cells demonstrated decreased proliferation and invasion and increased apoptosis （P<0.01）. ConclusionPTPN1 may be one of the core targets of JYP in the treatment of HCC. It is highly expressed in the HCC tissue and cells， which is associated with the poor prognosis of patients. The expression level of PTPN1 is significantly up-regulated in the HCC tissue of the patients with TP53 mutation. However， TP53 mutation or deletion does not affect the expression of PTPN1 in HCC cells. JYP can significantly down-regulate the expression of PTPN1 to inhibit the proliferation and invasion and promote the apoptosis of HCC cells.

Hepatic stellate cells (HSCs) are the primary fibrogenic cells in the liver, and their activation plays a crucial role in the development and progression of hepatic fibrosis. Here, we report that retinoid X receptor-alpha (RXRα), a unique member of the nuclear receptor superfamily, is a key modulator of HSC activation and liver fibrosis. RXRα exerts its effects by modulating calcium/calmodulin-dependent protein kinase kinase β (CaMKKβ)-mediated activation of AMP-activated protein kinase-alpha (AMPKα). In addition, we demonstrate that K-80003, which binds RXRα by a unique mechanism, effectively suppresses HSC activation, proliferation, and migration, thereby inhibiting liver fibrosis in the CCl₄ and amylin liver NASH (AMLN) diet animal models. The effect is mediated by AMPKα activation, promoting mitophagy in HSCs. Mechanistically, K-80003 activates AMPKα by inducing RXRα to form condensates with CaMKKβ and AMPKα via a two-phase process. The formation of RXRα condensates is driven by its N-terminal intrinsic disorder region and requires phosphorylation by CaMKKβ. Our results reveal a crucial role of RXRα in liver fibrosis regulation through modulating mitochondrial activities in HSCs. Furthermore, they suggest that K-80003 and related RXRα modulators hold promise as therapeutic agents for fibrosis-related diseases.

BACKGROUND:There is no consensus on the optimal bone tunnel position in the lateral clavicle,which guides coracoclavicular ligament reconstruction.Postoperative complications such as enlargement of the lateral clavicle bone tunnel,bone osteolysis,clavicle fracture,and failure of internal fixation are likely to occur.Bone mass density plays an important role in the strength and stability of endophytic fixation.Regional differences in the bone mass density of the distal clavicle should not be overlooked in the repair and reconstruction of acromioclavicular dislocation.Currently,there are no quantitative clinical studies in humans regarding the bone mass density of the distal clavicle. OBJECTIVE:To measure the magnitude of bone mass density in different regions of the distal clavicle by quantitative CT to provide a reference for surgeons to repair and reconstruct the coracoclavicular ligament. METHODS:101 patients undergoing quantitative CT checking in Fuyang People's Hospital Affiliated to Anhui Medical University from October to December 2022 were enrolled,from which 1 616 samples of subdivisional bone mass density of the distal clavicle were measured.For each of the quantitative CT samples,firstly,the distal clavicle was divided medially to laterally into the following four regions:conical nodal region(region A),inter-nodal region(region B),oblique crest region(region C)and distal clavicular region(region D).Secondly,each region was divided into the first half and the second half to determine eight subdivisions,then setting semiautomatic region of interest(ROI)in each subdivision:(ROI A1,A2,B1,B2,C1,C2,D1,and D2).Thirdly,each quantitative CT scan was transferred to the quantitative CT pro analysis workstation,and cancellous bone mass density was measured in the distal clavicle ROI.Finally,the clavicular cortex was avoided when measuring. RESULTS AND CONCLUSION:(1)There was no statistically significant difference in bone mineral density on the different sides of the shoulder(P＞0.05).(2)The analysis of bone mineral density in eight sub-areas of the distal clavicle A1,A2,B1,B2,C1,C2,D1,and D2 showed statistically significant differences(P＜0.05).It could be considered that there were differences in bone mineral density in different areas of the distal clavicle.After pairwise comparison,there was no statistically significant difference in bone mineral density between A1 and A2,D1 and D2,A2 and B1(P＞0.05),and there was a statistically significant difference in bone mineral density between the other sub-areas(P＜0.05).(3)The bone mineral density in the region A2 of the anatomical insertion of the conical ligament was significantly higher than that in the inter-nodular area(region B)(P＜0.05).The bone mineral density in the region A1 was higher than that in the region A2,but the difference was not statistically significant(P＞0.05).The bone mineral density in the region C1 of the anatomical insertion of the trapezium ligament was higher than that in regions C2,D1 and D2,and the bone mineral density in the inter-nodular area(region B)was significantly higher than that in regions C and D(P＜0.05).(4)These results have suggested that there are differences in bone mass density in different regions of the distal clavicle;regional differences in bone mass density in the distal clavicle during repair and reconstruction of acromioclavicular dislocation cannot be ignored.Consideration should be given not only to biomechanical factors but also to the placement of implants or bone tunnels in regions of higher bone mass density,which could improve the strength and stability of implant fixation and reduce the risk of complications such as bone tunnel enlargement,osteolysis,fracture and implant failure.

African swine fever(ASF),caused by the African swine fever virus(ASFV),is a highly contagious infectious disease of pigs.This disease has been spread rapidly in China since 2018,po-sing a huge threat to China's pig farming industry.To rapid detect the ASFV,a loop-mediated iso-thermal amplification(LAMP)combined with the disposable nucleic acid visualization test strip was established for visual detection of the nucleic acid of ASFV B646L gene.The method was easy to operate without special instruments and equipment,while it effectively avoided the disadvantage of false positives caused by aerosol contamination.The method was able to detect 1.16 copies/μL of the recombinant plasmid in 50 min at 65 ℃.In addition,the method was specific with no cross-re-action with classical swine fever virus,porcine reproductive and respiratory syndrome virus,por-cine parvovirus,transmissible gastroenteritis virus.The results in this study provides a rapid,con-venient,sensitive and reliable method for early diagnosis and screening for ASFV suspected infec-tion cases.

Objective To explore the value of gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid(Gd-EOB-DTPA)enhanced MRI T1 mapping,R2*quantitative parameters,and their combined indicators in assessing liver function in patients with hepatitis B cirrhosis.Methods The data from 52 patients with hepatitis B cirrhosis who underwent Gd-EOB-DTPA enhanced MRI and modified Dixon quantification(mDixon-Quant)scans were analyzed retrospectively.Patients were divided into three groups based on Child-Pugh scores:Child-Pugh A,B,and C.T1 mapping images were obtained pre,mid,and HBP.Quantitative values of T1 relaxation time reduction rates(△T1 mid and △T1 HBP),liver-related T1 relaxation times(T1 pre,T1 mid,and T1 HBP),fat fraction(FF),and R2* were measured and calculated.One-way ANOVA was used to compare the differences in MRI quantitative parameters between different Child-Pugh grading groups.Spearman's rank correlation analysis was performed to assess the correlation between MRI quantitative parameters and Child-Pugh grading.The receiver operating characteristic(ROC)curve was used to evaluate the performance of distinguishing liver function classification by statistically significant MRI parameters.Results Between different Child-Pugh grading groups,the differences in T1 mid,T1 HBP,△T1 mid,△T1 HBP,and R2*were statistically significant(P＜0.05),however T1 pre and FF were not statistically significant(P＞0.05).T1 mid,T1 HBP and R2*were positively correlated with Child-Pugh grading(rs=0.365,0.566,0.597,respectively;P＜0.05),meanwhile △T1 mid and△T1 HBP were negatively correlated with Child-Pugh grading(rs=-0.680,-0.771,respectively;P＜0.05).There were no significant correlations between T1 pre,FF and Child-Pugh grading(P＞0.05).The area under the curve(AUC)of T1 HBP,△T1 mid,△T1 HBP,R2*and their combined indicators for distinguishing Child-Pugh A grade from Child-Pugh B grade were about 0.888,0.784,0.955,0.764,and 0.961,respectively(P＜0.05).The AUC of △T1 mid,△T1 HBP,R2* and their combined indicators for distinguishing Child-Pugh B grade from Child-Pugh C grade were about 0.853,0.860,0.797,and 0.941,respectively(P＜0.05).Conclusion Gd-EOB-DTPA enhanced MRI T1 mapping and R2*quantitative parameters can be used to independently evaluate the status of liver function in hepatitis B cirrhosis,and the combined evaluation of the two kinds of parameters has a higher diagnostic efficiency.

OBJECTIVE: To improve the diagnostic and therapeutic efficiency for secondary laryngeal tuberculosis through an analysis on the clinical features of patients with this disease. METHOD: A retrospective study was made among 49 cases with laryngeal tuberculosis treated in Tibetan General Hospital of Chinese PLA, and the clinical data were carefully analyzed to summarize the clinical experience of this disease. RESULT: Of 49 patients, 24 cases had 1 year history, 11 cases had 1 to 3 years, 9 cases had 3 to 5 years, 5 cases had 5 years or more. Thirty-eight patients had the history of tuberculosis and 11 had none. Thirty-four patients had taken anti-tuberculosis drugs but none had standard therapy as demanded. All cases had mild general symptoms (mild fever, night sweats, weight loss, et al) and atypical local symptoms (hoarseness, sore throat). Therefore, 42 cases were misdiagnosed as non-specific chronic laryngitis, of which 15 cases got worse after oral administration or inhaling of steroid hormones. Seven persons were misdiagnosed as laryngeal cancer. All patients were confirmed pulmonary tuberculosis by X ray exam or CT scanning. Twelve cases had strong positive PPD tests and 2 cases were detected positive by sputum smear. All patients was treated by standard systematic and local chemical therapy against tuberculosis (inhaling of antituberculosis drugs for 1 to 2 months). All were cured but one died in a road accident, and none had recurrence after 1- to 9- year follow-up. CONCLUSION All of those the patients with long period hoarseness and sore throat should take chest CT scan or X-ray exam for the highest incidence of pulmonary tuberculosis at high altitudes. CT scanning is the prefer for its high resolution. Pathological biopsy and diagnostic therapy should be taken to make accurate diagnosis. Usually steroid hormones should not be recommended.
Adolescent ; Adult ; Aged ; Altitude ; Child ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Tibet ; Tuberculosis, Laryngeal ; diagnosis ; drug therapy ; Young Adult

Objective An effective reproducible protocol for complete plant regeneration via somatic embryogenesis has beendeveloped for Herpetospermum pedunculosum,an endangered Tibetan medicinal herb.Methods The cotyledonexplants used in this study were excised from seedlings germinated in vitro.Callus was induced from cotyledonexplants on Murashige and Skoog's medium,supplemented with 2,4-dichlorophenoxyacetic acid(2,4-D,0.1-1.0mg/L)alone or in combination with 6-benzylaminopurine(BA,0.5,1.0,and 2.0 mg/L).Results The calli showeddifferentiation of globular embryos after three weeks of incubation on MS medium supplemented with variouscombinations of BA and NAA.Sixty-two percent of the embryogenic calli produced somatic embryos in MS basalmedium supplemented with BA(1.0 mg/L)+NAA(2.0 mg/L).The addition of KN(0.5 mg/L)to MS mediumcontaining both BA and NAA(2.0 mg/L each)significantly increased the frequency of somatic embryogenesis.Themaximum percentage of embryogenic calli formation was 83%,and globular embryos formed and germinatedsuccessfully in this medium.Then,transferring the regenerated plants from this medium to hormone-free MSmedium will further enhanced the development of the plants,and the healthy plantlets are formed successfullywithin four weeks.The plantlets were transferred to soil to acclimatize under greenhouse conditions and 75%survived.Conclusion Somatic embryogenesis protocol as reported here can play a key role in the propagation andconservation of this endangered species.

OBJECTIVE:To assess the clinical efficacy and safety of domestic sparfloxacin for acute bacterial infections.METHODS:Domestic literatures about sparfloxacin for acute bacterial infections were retrieved by computer and their quality was evaluated to extract data(1993～2009).RevMan 4.2.2 software was used for Meta-analysis.RESULTS:A total of 10 RCT were enrolled.The comparisons of 2 groups were homogeneous in terms of clinical cure rate,clinical response rate,bacterial clearance rate and safety.There was statistical significance in comparison of combined effect variable between 2 groups in respect of cure rate,response rate and bacterial clearance rate(P0.05).CONCLUSION:The currently available evidence shows that clinical efficacy of domestic sparfloxacin for acute bacterial infections is better and incidence of ADR was lower.