OBJECTIVE To explore and predict the quality markers （Q-markers） of Compound xiebai capsules for the treatment of chronic obstructive pulmonary disease （COPD） by constituents analysis combined with network pharmacology and molecular docking studies， and to establish the quality standard of Compound xiebai capsules. METHODS UHPLC-TOF-MS was used for qualitative analysis of Compound xiebai capsules， and the candidate Q-markers of Compound xiebai capsules were screened by combining network pharmacology and molecular docking technology. Further， HPLC was applied to establish the fingerprints of 15 batches of Compound xiebai capsules and to conduct quantitative analysis of the main components. RESULTS A total of 51 components were identified from Compound xiebai capsules. Among them， 15 components， namely oxyberberine， methylworenine， coptisine， tetrahydroberberine， epiberberine， berberine， magnoflorine， gandensin， cucurbitacin D， hydroxygenkwan， jatrorrhizine， columbamine， quercetin， cucurbitacin R， and palmatine， were determined as the candidate Q-markers for Compound xiebai capsules in the treatment of COPD. A total of 13 common peaks were calibrated in the fingerprints of 15 batches of Compound xiebai capsules for COPD treatment， with similarity values ranging from 0.976 to 0.999 compared to the reference fingerprint. Seven components were identified among these peaks， namely peak 5 （magnoflorine）， peak 8 （jatrorrhizine）， peak 9 （epiberberine）， peak 10 （columbamine）， peak 11 （coptisine）， peak 12 （palmatine）， and peak 13 （berberine）. Their respective contents were （0.267±0.048）， （0.453±0.084）， （0.572±0.160）， （0.392±0.074）， （1.076±0.273）， （1.477±0.271）， and （6.664±1.249） mg/g （ n ＝3）. CONCLUSIONS This study predicted 15 candidate Q-markers of Compound xiebai capsules in the treatment of COPD and established the fingerprint along with a quantitative determination method for seven major components.

ObjectiveTo investigate the possible mechanism of Pibai Yucuo Formula （枇柏愈痤方， PYF） in treating acne from the perspective of skin barrier damage. MethodsThirty-two mice were randomly divided into blank group， model group， minocycline group， and PYF group， with 8 mice in each group. Except for the blank group， mice were induced by intradermal injection of Cutibacterium acnes （C.acnes） combined with topical application of artificial sebum to establish acne model. The blank group and model group received intragastric administration of 0.2 ml of distilled water， while the PYF group received intragastric administration of 22.75 g/（kg·d）of PYF， and the minocycline group received 0.013 g/（kg·d）of minocycline suspension， all once daily for 5 consecutive days. On day 0 and day 6 of the experiment， the body weight of mice in each group was recorded， and the absolute value of the body weight difference during the experiment was calculated. Skin conditions were assessed with multifunctional skin imaging system on the 2nd， 4th and 6th day of the experiment. Skin barrier function indicators including transepidermal water loss （TEWL）， and the water content of the stratum corneum and epidermis on day 0， 2， 4 and 6 of the experiment. Optical coherence tomography （OCT） was used to observe stratum corneum and skin thickness on the 1st， 3rd and 5th day of the experiment. Hematoxylin-eosin （HE） staining was performed to observe histopathological changes， while ELISA was used to detect interleukin-17A （IL-17A） levels， and immunofluorescence staining was used to assess skin barrier-related proteins filaggrin （FLG） and loricrin （LOR） levels of skin lesions on day 6 of the experiment. ResultsCompared to the blank group， the model group showed a decrease in body weight on day 6， and an increase in the absolute value of the difference in body weight before and after the experiment （P＜0.05）. On day 4 and 6， TEWL values increased， while water content in the skin stratum corneum and epidermis decreased （P＜0.05）， accompanied by elevated IL-17A level and reduced immunofluorescence intensity of FLG and LOR proteins （P＜0.05）. The model group mice showed papules or pustules at the skin modeling site with progressively worsening desquamation under multifunctional skin imaging system. OCT revealed focal epidermal protrusions， blurred epidermal-dermal boundaries， and disorganized structural layers. HE staining showed significant epidermal hyperkeratosis and incomplete keratinization in the skin， with keratin plug formation in hair follicles and glandular lumens， thickened stratum corneum， hyperplasia of the stratum spinosum， as well as dense dermal inflammatory cell infiltration， and capillary dilation. Compared to the model group， both the minocycline group and the PYF group showed a reduced difference in body weight before and after experiment （P＜0.05）. On day 4 and 6， the TEWL value decreased， and water content of the skin stratum corneum increased （P＜0.05）； on day 6， the IL-17A level in the skin lesions decreased and immunofluorescence intensity of FLG and LOR proteins increased （P＜0.05）. On day 4 and 6， the severity of the skin lesions and range of redness and swelling were lighter than those in the model group， with reverted epidermal thickness， smoother surface and clearer epidermis-dermis boundary. HE staining showed that the degree of skin keratinization was reduced， and the inflammatory infiltration and vascular dilation in the dermis were improved compared to the model group. The PYF group showed better results than the minocycline group in reducing TEWL value on day 4 （P＜0.05）. ConclusionPYF may improve inflammation and skin barrier damage by downregulating IL-17A levels in lesion tissue and increasing skin barrier-related proteins， which could be one of the potential mechanism of action on acne.

Objective To explore the diagnostic value of the phenomenon of protein-cell separation in cerebrospinal fluid(CSF),as well as the single and combined detections of CSF immunoglobulin G(CSF-IgG),CSF albumin(CSF-ALB)and serum immunoglobu-lin G(S-IgG)for Guillain-Barre syndrome(GBS).Methods A retrospective analysis was conducted on the clinical data of 65 GBS patients(GBS group)and 65 patients with other neurological diseases(non-GBS group).CSF and serum samples were collected from both the groups to analyze the diagnostic efficacy of protein-cell separation phenomenon in CSF,as well as the single and combined de-tections of CSF-IgG,CSF-ALB,and serum-IgG(S-IgG)in GBS.Results CSF protein-cell separation phenomenon was observed in 60.00％(39/65)of GBS patients.Prodromal events were presented in 67.69％(44/65)of GBS patients,which mainly were upper re-spiratory tract infection(43.08％,28/65),digestive tract infection(9.23％,6/65),and herpetic virus infection(7.69％,5/65).The levels of CSF-IgG,CSF-ALB,and S-IgG in the GBS group were significantly higher than those in non-GBS group(P＜0.05).The areas under the ROC curves(AUCROC)for single CSF-IgG,CSF-ALB,S-IgG,protein-cell separation phenomenon IN CSF,and the combination of all the four parameters in diagnosis of GBS were 0.754,0.705,0.682,0.708,and 0.840,respectively.The diagnostic specificities were 87.7％,75.4％,87.7％,81.5％,and 96.9％,respectively.Conclusion Most of the GBS patients were found to have CSF protein-cell separation phenomenon and history of prodromal infection.CSF-IgG,CSF-ALB,S-IgG,and CSF protein-cell separation phenomenon exhibited independent diagnostic value for GBS.The combined detection of the four indicators could improve the diagnostic efficacy for GBS.

Objective To evaluate the morphological characteristics of dynamic contrast-enhanced magnetic resonance imaging(DCE-MRI)and diffusion-weighted imaging(DWI),and the diagnostic performance of quantitative and qualitative parameters in the diagnosis of neoadjuvant chemotherapy for breast cancer.Methods The database of PubMed,EMBASE,Cochrane Library,Web of Sci-ence,CNKI and Wanfang Data Knowledge Service Platform databases were searched.The search period was from January 2014 to July 2024.QUADAS-2 was used to evaluate the methodological quality of included studies.Data were extracted,and the data of true posi-tive,false positive,true negative and false negative were sorted out,and the sensitivity,specificity,positive predictive value(PLR)and negative predictive value(NLR)were calculated.Data merging,heterogeneity test,Meta regression analysis and subgroup analysis were performed using Stata 13.0 and RevMan 5.3.Results A total of 2244 breast cancer patients with an average age of 49.2 years were in-cluded in 17 literatures according to the inclusion and exclusion criteria.The sensitivity,specificity,PLR and NLR of DCE-MRI were 80％,75％,3.20,0.27,respectively.The sensitivity,specificity,PLR and NLR of DWI were 81％,64％,2.30,0.29,respectively.Subgroup analysis showed that magnetic field intensity had an effect on heterogeneity,but age,study design and different evaluation pa-rameters had no significant effect on heterogeneity.Conclusion There is no significant difference between DCE-MRI and DWI in the diagnostic efficacy of neoadjuvant chemotherapy for breast cancer.When patients are unable to undergo DCE-MRI,DWI can be a poten-tial non-invasive alternative examination.

Objective To invegtigate the possible etiology,clinical characteristics,and treatment effects of post tympanoplasty full-frequency bone conduction hearing loss in the operated ear.Methods A retrospective analy-sis was conducted on the possible causes,clinical manifestations,and diagnosis and treatment of 6 cases of full-fre-quency bone conduction hearing loss in the operated ear after tympanoplasty.Results The use of diluted epineph-rine during surgery,controlled hypotension during the operation,ethylene oxide in gelatin sponges,and the exces-sive pro-inflammatory factors produced postoperatively may be associated with the occurrence of full-frequency bone conduction hearing loss in the operated ear after tympanoplasty.Among the six patients,the main initial symptom was dizziness without vertigo or nystagmus.Two cases were ineffectively treated,one case was effectively treated,and three cases were cured.Among the four cases that received timely medication,three were effectively treated.Conclusion Full-frequency bone conduction hearing loss in the operated ear after tympanoplasty is related to various factors,and the clinical manifestations of patients are atypical,often manifesting as dizziness.Timely diagnosis through pure tone audiometry and timely treatment according to the sudden deafness treatment plan often leads to fa-vorable outcomes.

ObjectiveTo explore the possible action mechanism of Qingshi Anti-itch Ointment （青石止痒软膏， QAO） in the treatment of psoriasis. MethodsForty mice were randomly divided into four groups， blank group， model group， calcipotriol group and QAO group， with 10 mice in each group. Except for the blank group， psoriasis was induced by applying imiquimod cream to the dorsal skin. After modeling for 6 hours daily， the calcipotriol group and QAO group were treated with 0.5 g of calcipotriol ointment or 0.5 g of QAO， respectively， applied to the treated dorsal skin. The blank group and the model group received no treatment. The skin lesions were observed， and the psoriasis area and severity index （PASI） score was assessed every other day. After 7 days， Hematoxylin and Eosin （HE） staining was performed on dorsal skin tissue to observe pathological changes. The levels of interleukin 1β （IL-1β） and interleukin 18 （IL-18） were determined by enzym-linked immunosorbent assay （ELISA）. The protein levels of Caspase-1，Pro-Caspase-1， gasdermin D （GSDMD） and gasdermin-D-N （GSDMD-N） were detected by Western Blot （WB）. The protein levels of GSDMD were observed by immunohistochemistry. ResultsCompared with the blank group， the model group mice showed redness， erythema， and white scales on their skin， with histological observations indicating epidermal thickening， elongated spines， and infiltration of inflammatory cells. The PASI scores of the skin tissue on days 1， 3， 5， and 7 were elevated； the IOD and AOD values of GSDMD protein increased； the protein levels of Caspase-1， Pro-Caspase-1，GSDMD， GSDMD-N， and IL-1β and IL-18 were significantly elevated （P＜0.05 or P＜0.01）. Compared with the model group， the QAO group and calcipotriol group showed lighter skin lesions； the PASI scores on day 5 and day 7 in the QAO group， and on day 3， 5， and 7 in the calcipotriol group， were reduced； the IOD and AOD values of GSDMD protein， and the protein level of Caspase-1， GSDMD， and GSDMD-N， as well as level of IL-18 and IL-1β decreased in both groups； in the calcipotriol group， Pro-Caspase-1 protein level also decreased （P＜0.05 or P＜0.01）. Compared with the calcipotriol group， the QAO group showed slightly redder skin， more obvious thickening of the stratum corneum， and less capillary dilation； the PASI scores on day 3 and day 7 increased， while the score on day 5 was reduced； the protein level of Pro-Caspase-1， GSDMD， GSDMD-N， and the level of IL-18 and IL-1β were increased in the QAO group （P＜0.05）. ConclusionQAO can effectively relieve psoriasis dermatitis in mice. Its potential mechanism may be related to the regulation of the Caspase-1/GSDMD protein pathway， down-regulation of IL-18 and IL-1β levels， and alleviation of pyroptosis.

Objective To evaluate the morphological characteristics of dynamic contrast-enhanced magnetic resonance imaging(DCE-MRI)and diffusion-weighted imaging(DWI),and the diagnostic performance of quantitative and qualitative parameters in the diagnosis of neoadjuvant chemotherapy for breast cancer.Methods The database of PubMed,EMBASE,Cochrane Library,Web of Sci-ence,CNKI and Wanfang Data Knowledge Service Platform databases were searched.The search period was from January 2014 to July 2024.QUADAS-2 was used to evaluate the methodological quality of included studies.Data were extracted,and the data of true posi-tive,false positive,true negative and false negative were sorted out,and the sensitivity,specificity,positive predictive value(PLR)and negative predictive value(NLR)were calculated.Data merging,heterogeneity test,Meta regression analysis and subgroup analysis were performed using Stata 13.0 and RevMan 5.3.Results A total of 2244 breast cancer patients with an average age of 49.2 years were in-cluded in 17 literatures according to the inclusion and exclusion criteria.The sensitivity,specificity,PLR and NLR of DCE-MRI were 80％,75％,3.20,0.27,respectively.The sensitivity,specificity,PLR and NLR of DWI were 81％,64％,2.30,0.29,respectively.Subgroup analysis showed that magnetic field intensity had an effect on heterogeneity,but age,study design and different evaluation pa-rameters had no significant effect on heterogeneity.Conclusion There is no significant difference between DCE-MRI and DWI in the diagnostic efficacy of neoadjuvant chemotherapy for breast cancer.When patients are unable to undergo DCE-MRI,DWI can be a poten-tial non-invasive alternative examination.

ObjectiveTo evaluate the clinical efficacy and safety of Huaban Jiedu Formulation （化斑解毒方， HJF） in treating psoriasis vulgaris with blood-heat syndrome. MethodsA randomized， double-blind， placebo-controlled study was conducted with 60 patients diagnosed with psoriasis vulgaris of blood-heat syndrome. Patients were randomly assigned to either a treatment group or a control group， with 30 cases in each. The treatment group received HJF granules orally， one dose a day， combined with topical Qingshi Zhiyang Ointment （青石止痒软膏）， while the control group received placebo granules， one dose a day， combined with the same topical ointment. Both groups were topically treated twice daily of 28 days treatment cours. Psoriasis area and severity index （PASI）， visual analogue scale for pruritus （VAS）， traditional Chinese medicine （TCM） syndrome scores， dermatology life quality index （DLQI）， and psoriasis life stress inventory （PLSI） were assessed before treatment and on day 14 and day 28. Response rates for PASI 50 （≥50% reduction） and PASI 75 （≥75% reduction）， as well as overall clinical efficacy， were compared between groups. Serum levels of interleukin-6 （IL-6） and interleukin-17 （IL-17） were measured before and after 28 days of treatment. Adverse reactions during treatment were recorded. ResultsAfter 28 days of treatment， both groups showed significant reductions in PASI total score， lesion area score， erythema， scaling， and infiltration scores， pruritus VAS score， TCM syndrome score， DLQI， PLSI， and serum IL-6 and IL-17 levels （P＜0.05）. Compared to the control group， the treatment group had significantly greater improvements in PASI total score and erythema score， TCM syndrome score， serum IL-6 and IL-17 levels， and PASI 50 response rate after 28 days （P＜0.05）. Between-group comparisons of score differences before and after 28-day treatment revealed that the treatment group showed significantly better improvements in PASI total， lesion area score， erythema score， TCM syndrome score， DLQI， PLSI， and inflammatory markers （P＜0.05 or P＜0.01）. The total effective rate on day 14 and day 28 was 40.00% （12/30） and 83.33% （25/30） in the treatment group， versus 6.90% （2/29） and 41.38% （12/29） in the control group， respectively. The clinical efficacy in the treatment group was significantly superior to that in the control group （P＜0.05）. Mild gastric discomfort occurred in 3 patients in the treatment group and 1 in the control group. ConclusionHJF can effectively improve skin lesions and TCM symptoms relieve pruritus， enhance quality of life， and reduce inflammatory markers IL-6 and IL-17， in patients with blood-heat syndrome of psoriasis vulgaris， with a good safety profile.

Objective To explore the diagnostic value of the phenomenon of protein-cell separation in cerebrospinal fluid(CSF),as well as the single and combined detections of CSF immunoglobulin G(CSF-IgG),CSF albumin(CSF-ALB)and serum immunoglobu-lin G(S-IgG)for Guillain-Barre syndrome(GBS).Methods A retrospective analysis was conducted on the clinical data of 65 GBS patients(GBS group)and 65 patients with other neurological diseases(non-GBS group).CSF and serum samples were collected from both the groups to analyze the diagnostic efficacy of protein-cell separation phenomenon in CSF,as well as the single and combined de-tections of CSF-IgG,CSF-ALB,and serum-IgG(S-IgG)in GBS.Results CSF protein-cell separation phenomenon was observed in 60.00％(39/65)of GBS patients.Prodromal events were presented in 67.69％(44/65)of GBS patients,which mainly were upper re-spiratory tract infection(43.08％,28/65),digestive tract infection(9.23％,6/65),and herpetic virus infection(7.69％,5/65).The levels of CSF-IgG,CSF-ALB,and S-IgG in the GBS group were significantly higher than those in non-GBS group(P＜0.05).The areas under the ROC curves(AUCROC)for single CSF-IgG,CSF-ALB,S-IgG,protein-cell separation phenomenon IN CSF,and the combination of all the four parameters in diagnosis of GBS were 0.754,0.705,0.682,0.708,and 0.840,respectively.The diagnostic specificities were 87.7％,75.4％,87.7％,81.5％,and 96.9％,respectively.Conclusion Most of the GBS patients were found to have CSF protein-cell separation phenomenon and history of prodromal infection.CSF-IgG,CSF-ALB,S-IgG,and CSF protein-cell separation phenomenon exhibited independent diagnostic value for GBS.The combined detection of the four indicators could improve the diagnostic efficacy for GBS.

L-citrulline is a nonprotein amino acid that plays an important role in human health and has great market demand. Although microbial cell factories have been widely used for biosynthesis, there are still challenges such as genetic instability and low efficiency in the biosynthesis of L-citrulline. In this study, an efficient, plasmid-free, non-inducible L-citrulline-producing strain of Escherichia coli BL21(DE3) was engineered by combined strategies. Firstly, a chassis strain capable of synthesizing L-citrulline was constructed by block of L-citrulline degradation and removal of feedback inhibition, with the L-citrulline titer of 0.43 g/L. Secondly, a push-pull-restrain strategy was employed to enhance the L-citrulline biosynthesis, which realized the L-citrulline titer of 6.0 g/L. Thirdly, the NADPH synthesis and L-citrulline transport were strengthened to promote the synthesis efficiency, which achieved the L-citrulline titer of 11.6 g/L. Finally, fed-batch fermentation was performed with the engineered strain in a 3 L fermenter, in which the L-citrulline titer reached 44.9 g/L. This study lays the foundation for the industrial production of L-citrulline and provides insights for the modification of other amino acid metabolic networks.
Citrulline/biosynthesis* ; Escherichia coli/genetics* ; Metabolic Engineering/methods* ; Fermentation ; NADP/biosynthesis*