ObjectiveTo reveal the active substances and mechanisms of modified Qing'e formula （MQEF） in delaying skin photoaging by ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry （UPLC-Q-TOF-MS），network pharmacology， and cell experiments. MethodsUPLC-Q-TOF-MS and a literature review were employed to analyze the transdermally absorbed components in mice after the topical application of MQEF. The potential targets of MQEF in treating skin photoaging were retrieved from databases.The compound-potential target network and protein-protein interaction network were constructed to screen the key components and core targets. A photoaging cell model was established by irradiating HaCaT cells with medium-wave ultraviolet B （UVB）. The safe doses of bakuchiol （BAK） and salvianolic acid B （SAB） for treating HaCaT cells and the effects of BAK and SAB on the viability of cells exposed to UVB irradiation were determined by the cell counting kit-8 （CCK-8） method.The reactive oxygen species （ROS） fluorescent probe was used to measure the ROS production in the cells treated with BAK and SAB.The expression levels of genes related to oxidative stress，inflammation，collagen metabolism，and circadian rhythm clock were measured by Real-time PCR. ResultsA total of 24 transdermally absorbed components of MQEF were identified，which acted on 367 potential targets，and 417 targets related to skin photoaging were screened out，among which 47 common targets were predicted as the targets of MQEF in treating skin photoaging. MQEF exerted the anti-photoaging effect via key components such as BAK and SAB，which acted on core proteins such as serine/threonine kinase 1 （Akt1） and mitogen-activated protein kinase 3 （MAPK3） and intervened in core pathways such as the tumor necrosis factor （TNF） and phosphatidylinositol-3-kinase （PI3K）/protein kinase B （Akt） signaling pathways.Compared with the model group，the administration of BAK and SAB increased the survival rate of HaCaT cells （P<0.01），down-regulated the mRNA levels of cyclooxygenase-2 （COX-2），interleukin-6 （IL-6），tumor necrosis factor-α （TNF-α），matrix metalloproteinase-1 （MMP-1），and matrix metalloproteinase-9 （MMP-9） （P<0.01），and up-regulated the mRNA levels of heme oxygenase-1 （HO-1） and NAD（P）H quinone dehydrogenase 1 （NQO-1） （P<0.05，P<0.01） in photoaged HaCaT cells.In addition，it eliminated excess ROS production induced by UVB and up-regulated the mRNA levels of brain and muscle ARNT-like 1 （BMAL1） and circadian locomotor output cycles kaput （CLOCK） associated with circadian clock （P<0.05，P<0.01）. ConclusionMQEF delays skin photoaging through the coordinated effects of various components，multiple targets，and diverse pathways.The key components BAK and SAB in MQEF exhibit anti-photoaging properties，which involve inhibiting oxidative stress，preventing collagen degradation，mitigating inflammation，and maintaining normal skin circadian rhythms by regulating clock gene expression.

OBJECTIVE To establish the fingerprint of Pinghuo tea standard decoction and a method for determination of multi-component to clarify the transfer relationship of quantities and quality from pieces and standard decoction. METHODS Fifteen batches of Pinghuo tea standard decoction were prepared and the extract rate was determined； the fingerprint of the preparation was established by using high-performance liquid chromatography（HPLC）； the similarity evaluation and the determination of common peaks were performed， and chemometric analysis was performed； the same method was used to determine the content of indicator components and the transfer rate was calculated. The chromatographic column was Venusil C18 column with mobile phase consisted of acetonitrile-0.1% phosphoric acid solution （gradient elution）； the column temperature was 30 ℃， and the detection wavelengths were 238 nm （0-37 min， 85-102 min） and 330 nm （37-85 min） at a flow rate of 1.0 mL/min with an injection volume of 10 μL. RESULTS The similarity of HPLC fingerprints for 15 batches of Pinghuo tea standard decoction was not lower than 0.968. A total of 24 common peaks were calibrated and 9 peaks were recognized， which were as follows neochlorogenic acid （peak 3）， chlorogenic acid （peak 6）， geniposide （peak 9）， glycyrrhizin （peak 10）， galuteolin （peak 11）， isochlorogenic acid A （peak 14）， luteolin （peak 21）， kaempferol （peak 23） and glycyrrhizic acid （peak 24）. Cluster analysis， principal component analysis and orthogonal partial least squares discriminant analysis showed consistent results， all of which could classify the 15 batches of samples into three categories. The linear range of indicator components in 15 batches of Pinghuo tea standard decoction， such as geniposide， luteolin， isochlorogenic acid A， glycyrrhizin， and glycyrrhizic acid， were 0.020 580-0.411 600， 0.001 617-0.080 850， 0.006 076-0.607 600， 0.005 125-0.071 740， and 0.017 288-0.432 200 mg/mL， respectively； RSDs of precision， repeatability， stability and recovery rate tests were all not higher than 4% （n＝6）. The mass fractions ranged 3.227 9-10.002 2， 0.297 4-0.554 6， 3.350 1-6.159 6， 0.720 6-1.073 3， 2.003 1-3.030 1 mg/g； transfer rates from the pieces and standard decoction were 19.762 8%-35.840 5%， 12.123 3%-21.254 0%， 46.097 2%-82.869 4%， 58.708 8%-91.629 6%， 39.114 3%-63.710 6%. The transfer rates of the extract from 15 batches of Pinghuo tea standard decoction ranged from 61.15%-84.68%. CONCLUSIONS Established HPLC fingerprint and content determination methods in this study are simple and accurate， which can provide reference for the quantitative value transfer study， quality control， clinical application and the development of subsequent formulations of Pinghuo tea standard decoction.

AIM:To investigate the ameliorative effect of nociceptin/orphanin(N/OFQ)on anxiety-like be-havior in nicotine(NIC)withdrawal-induced rats and its regulatory mechanisms on the expression of neurotransmitters as-sociated with the hypothalamic-pituitary-adrenal(HPA)axis and inflammatory factors.METHODS:Thirty-two adult male Sprague-Dawley rats were randomly divided into 4 groups:normal control group,NIC withdrawal model group,low-dose N/OFQ treatment group,and high-dose N/OFQ treatment group,with 8 rats in each group.To establish NIC with-drawal model,the rats in the NIC model and N/OFQ treatment groups were subcutaneously injected with NIC(0.4 mg/kg),twice a day for 7 consecutive days followed by 3 days of withdrawal.During the withdrawal period,the rats in the low and high-dose N/OFQ treatment groups received intracerebroventricular injection of N/OFQ(1 nmol or 10 nmol)once per day for 3 consecutive days.Ten minutes after the third administration,all rats underwent open filed(OF)and elevated plus maze(EPM)tests to detect behavioral changes.The serum concentrations of corticotrophin-releasing hormone(CRH),adrenocorticotropic hormone(ACTH),corticosterone(CORT),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and IL-6 were measured by ELISA.The mRNA expression levels of TNF-α,IL-1β and IL-6 in the central nu-cleus of the amygdala(CeA)of the brain were detected by RT-qPCR.Histological changes in neuron morphology in the CA1 region of the hippocampus were observed under a light microscope following hematoxylin and eosin(HE)staining.Norepinephrine(NE)levels in the CeA of the brain were determined by HPLC.The protein expression of tyrosine hydroxy-lase(TH)in the CeA of the brain was detected by Western blot.RESULTS:Compared with the NIC withdrawal model group,rats in the low and high-dose N/OFQ treatment groups showed significant increase in the distance and time spent in the central area of the open field(P<0.05 or P<0.01),as well as significant increase in the number of entries and the per-centage of time spent in the open arms of the EPM(P<0.05 or P<0.01).Furthermore,both low and high-dose N/OFQ treatment groups significantly inhibited serum concentrations of CRH,ACTH and CORT in NIC withdrawal rats(P<0.01).N/OFQ administration also significantly reduced the levels of inflammatory cytokines TNF-α,IL-1β,and IL-6 in the serum,as well as expression levels of TNF-α,IL-1β and IL-6 mRNA in the CeA(P<0.05 or P<0.01).The treatment with N/OFQ at both doses significantly alleviated neuronal damage in the CA1 region of the hippocampus and markedly re-duced thelevels of NE and TH protein expression in the CeA of NIC withdrawal rats(P<0.01).CONCLUSION:N/OFQ alleviates anxiety-like behavior in NIC withdrawal rats through mechanisms related to the regulation of HPA axis hormone levels and inflammatory factors.

Objective To observe the effect of extracorporeal shockwave therapy (ESWT) in regulating endothelial cell phosphatase and tensin homolog deleted on chromosome ten (PTEN) expression on lower ex-tremity vascular lesion and its possible mechanism.Methods Twenty-four 2-month-old healthy male SD rats were randomly divided into the three groups:control group (group A),diabetes angiopathy group (group B),diabetes angiopathy+ESWT group (group C).The group B and C were fed with high fat and high sugar and intraperitoneally injected with streptozotocin 60 mg/kg to establish the rat model of diabetes vascular lesion. The group C received ESWT at 1 week (T1),2 weeks (T2),3 weeks (T3) and 4 weeks (T4) after modeling,and the blood stream velocity of rat femoral artery vascular lesion area and vascular internal diameter were measured at T4 by ultrasound.At the end of ESWT,the rats were immediately killed for taking their femoral arteries and gastrocnemius.The structures of the femoral arteries in each group were observed under electron microscopy.Western blot was used to detect the expression levels of PTEN,PI3K and Akt proteins,while qRT-PCR was used to detect the expression levels of PTEN mRNA.Immunofluorescence was used to detect the expression level of CD31 in gastrocnemius muscle .Results The peak systolic flow velocity and end-dias-tolic flow velocity of femoral artery at T4 in group B and C were significantly lower than those in group A (P<0.05),but group C was higher than group B (P<0.05).The internal diameter of femoral artery had no statistical difference among 3 groups (P>0.05).The PTEN expression level in group B and group C was sig-nificantly lower than that in group A (P<0.05),while group C was higher than group B (P<0.05).The ex-pression levels of PI3K and Akt in group B were higher than those in group A (P<0.05),and group C was lower than group B (P<0.05).The PTEN mRNA expression level in group B and group C was significantly lower than that in group A (P<0.05),but group C was higher than group B (P<0.05).Under electron mi-croscopy,it was observed that after ESWT,the endothelial cell damage in group C was obvious when com-pared with group B.The CD31 expression level in group B and group C was significantly lower than that in group A (P<0.05),but group C was higher than group B (P<0.05).Conclusion ESWT could improve the vascular function,increase the peak velocity during systolic period of femoral artery in diabetes rats and im-prove the microvessel density of gastrocnemius muscle by up-regulating PTEN in lower extremity artery and down-regulating PI3K and Akt in diabetes rats.

Objective To investigate the relationship between spicy diet and uremic pruritus(UP)in the patients with maintenance haemodialysis(MHD).Methods A total of 403 patients receiving the treat-ment in the blood purification center of this hospital from December 2023 to February 2024 were selected as the study subjects and grouped by the sum of the scores of frequency and degree of pepper intake.The visual analogue rating scale(VAS)was used to conduct the preliminary pruritus score in all patients,and the pa-tients with the score＞0 point conducted the multidimensional evaluation by the 14-item uremic skin pruritus scale.The blood routine and itch-related blood biochemical indexes levels of all patients were measured.Results There were 65 cases in the bland diet group,119 cases in the mild spicy diet group and 219 cases in the spicy diet group,and there was no significant difference in the number of genders between the groups(P＞0.05).There were statistically significant differences in age,dialysis age and lymphocyte count among the groups(P＜0.05).There was no statistically significant difference in the degree of pruritus among the groups(Z=9.301,P=0.157),but it was seen that the proportion of moderate and severe pruritus in the mild spicy diet group and the spicy diet group was decreased,and the proportions of no pruritus and mild pruritus showed the increasing trend.The itching score of the bland diet group was higher than that of the mildly spicy diet group and spicy diet group,and the difference was statistically significant(P＜0.05),but there was no statistically significant difference between the mild spicy diet group and spicy diet group(P＞0.05).There was a statistically significant difference in the itch score of the patients aged 40-60 years in each group(P＜0.05),and there was no statistically significant difference in the itch score between the patients aged＞60 years old and＜40 years old in each group(P＞0.05).There was no statistically significant difference in the itch-related blood biochemical indexes among the groups(P＞0.05).Conclusion The spicy diet may reduce the degree of pruritus in patients with MHD,moreover which is not affected by the age and other factors,and may be associated with lymphocyte level decrease in the patients.

BACKGROUND:Resistance to the inflammatory response is an important part of promoting the repair of damaged tissue and improving the local inflammatory response caused by medical bio-implant materials has been a key issue to be addressed in recent years. OBJECTIVE:To summarize the anti-inflammatory effects of common metal ions and related molecular mechanisms to provide some theoretical references for improving the early inflammatory response of hosts caused by bio-implant materials. METHODS:A computer search of the relevant literature in PubMed,Web of Science,CNKI and WanFang databases was conducted using"metal ions,magnesium ion,zinc ion,silver ion,copper ion,inflammation,anti-inflammatory effects,oxidative stress,immunoregulation,signaling pathways"as Chinese and English search terms.Preliminary screening was conducted by reading the titles and abstracts.Finally,80 papers were included for result analysis and summary. RESULTS AND CONCLUSION:(1)Metal ions such as magnesium,zinc,silver and copper have a good anti-inflammatory effect.The strength of this anti-inflammatory effect is strongly correlated with the dose and duration of action.In the future,consideration can be given to controlling the release rate of ions and adjusting the appropriate therapeutic concentration to achieve the best anti-inflammatory effect.(2)Magnesium ions and zinc ions exhibit excellent anti-inflammatory activity,with magnesium ions often being beneficial in anti-inflammatory therapy in the form of compounds such as magnesium sulfate and zinc ions regulating the body's inflammatory response with zinc feed as the main source of zinc supplementation.(3)Silver and copper ions have some anti-inflammatory effects,but are still predominant for their excellent antibacterial activity,mainly in the form of nanoparticles and bio-coatings.(4)Magnesium and zinc metal ions can be combined with natural extracts to form complexes to exert anti-inflammatory effects,and this method has the advantage of being inexpensive and widely available and is a sustainable and green approach,which is worthy of clinical promotion.(5)Metal ions such as magnesium,zinc,silver and copper exert anti-inflammatory effects by reducing host oxidative stress damage,modulating immune cells and inhibiting inflammatory signaling pathways such as nuclear factor-κB,Toll-like receptor,STAT3 and NOD.(6)The molecular mechanism related to the anti-inflammation of metal ions is a complex network,which is not the effect of a single pathway,but should be a combination of multiple signaling pathways.There are still many potential mechanisms that have not yet been explored,and more systematic elucidation of the interconnections between various signaling pathways is needed in the future.

Objective To investigate the effects of long noncoding RNA(lncRNA)-NRON on apoptosis following myocardial infarc-tion(MI)in mice.Methods The C57BL/6 mice were randomly divided into four groups:sham operation(Sham)group,MI group,MI combined with lncRNA-NRON interference lentivirus(MI+shNRON)group,and MI combined with the negative control(NC)lentivirus(MI+NC)group.The expression of lncRNA-NRON was detected using real-time PCR.In addition,the pathology of the myocardial tissue injury was analyzed using HE staining,the myocardial infarction size was examined using TTC staining,and the extent of apoptosis was assessed using the TUNEL assay,respectively.The RPISeq database was used to predict the probability of interaction between lncR-NA-NRON and the voltage-dependent anionic channel protein(VDAC).The effect of lncRNA-NRON on the expression of VDAC protein was detected using Western blotting.Results The lncRNA-NRON expression was significantly increased in the MI group,and the tar-geted knockdown of lncRNA-NRON resulted in alleviation of the pathological myocardial tissue injury,reduction in the myocardial infarc-tion area,and inhibition of apoptosis.The probability of interaction between lncRNA-NRON and VDAC reached 0.9,indicating a high probability of their association.Additionally,lncRNA-NRON could regulate the protein expression of VDAC.Conclusion Knockdown of lncRNA-NRON could reduce the occurrence of myocardial injury following myocardial infarction.This effect may be attributable to a spe-cific mechanism wherein lncRNA-NRON affects the process of apoptosis by binding to VDAC,consequently suppressing its expression.

Background Drinking water contains a variety of chemicals that may pose certain health risks to the human body. Objective To evaluate carcinogenic and non-carcinogenic health risks of chemical pollutants in drinking water in Nanjing from 2014 to 2022. Methods According to the Standard examination methods for drinking water (GB/T 5750-2006) and the Standards for drinking water quality (GB 5749-2006), the conventional water quality indexes of finished water, tap water, and secondary water supply in Nanjing from 2014 to 2022 were monitored. The health risk assessment model recommended by the United States Environmental Protection Agency (US EPA) was used to assess the health risks of 16 chemicals [arsenic, cadmium, chromium (hexavalent), lead, mercury, selenium, cyanide, fluoride, nitrate nitrogen, trichloromethane, carbon tetrachloride, aluminum, iron, manganese, copper, and zinc] in drinking water through different routes (drinking water and skin contact) in different populations (adult males, adult females, and children). Region (urban and rural), water period (dry period and wet period), and water sample type (finished water, tap water, and secondary water supply) were stratified for analysis. Results From 2014 to 2022, a total of 4198 samples of drinking water were monitored in Nanjing, including 483 samples of finished water, 3313 samples of tap water, and 402 samples of secondary water supply. The pass rates of all indicators were above 99%. The health risk assessment results showed that the carcinogenic/non-carcinogenic risks of arsenic and lead presenting a trend of first increasing from 2015 and 2016 respectively, remaining relatively high for 3 and 4 consecutive years respectively, and then decreasing from 2018 and 2020 respectively before remaining stable; the carcinogenic risk of cadmium was much higher than that of other chemicals, and the risk was greater than 1.00×10⁻⁴ except for 2018 and 2019. The non-carcinogenic risks of drinking water in different populations (adult males, adult females, and children) under different exposure modes (intake of drinking water and skin contact) were less than 1. The values of carcinogenic risk via drinking water in total population and of cadmium intake via drinking water in all sub-groups (adult males, adult females, and children) were greater than 1.00×10⁻⁴. The carcinogenic/non-carcinogenic risk of drinking water was the highest in children, and the carcinogenic/non-carcinogenic risk via drinking water ingestion was higher than that via skin contact. The stratified analysis showed that the carcinogenic/non-carcinogenic risks of arsenic and cadmium in urban drinking water were higher than those in rural drinking water (P<0.05); the carcinogenic/non-carcinogenic risks of arsenic and trichloromethane in wet season were higher than those in dry season (P<0.05); the carcinogenic risks of cadmium in different types of water were all greater than 1.00×10⁻⁴; the health risks varied by chemicals and water sample types for the other chemicals, but all were within an acceptable level. Conclusion The non-carcinogenic risks and carcinogenic risks of selected chemical pollutants in drinking water in Nanjing City from 2014 to 2022 show decreasing or stable trends. Among the 16 chemicals, the carcinogenic risk of cadmium exceeds the acceptable range recommended by the US EPA, and the carcinogenic/non-carcinogenic risks of the remaining chemicals are within the acceptable ranges. Cadmium should be given priority attention in future risk management of drinking water.

Objective:To establish an automated labeling method of 18F-AlF-1, 4, 7-triazocyclohexane-1, 4, 7-triacetic acid- D-Phe1-Tyr3-Thr8-octreotide (NOTATATE) and perform neuroendocrine tumor (NET) imaging. Methods:Based on the GE-FASTLab2 synthesis module, 18F-AlF-NOTATATE was automatically prepared by one-step chelation labeling with aluminum fluoride, and its labeling conditions were optimized. The product quality was analyzed. One patient (male, 47 years old) with lower rectal segment NET and one patient (female, 52 years old) with pancreatic NET underwent 18F-AlF-NOTATATE PET/CT imaging. Results:18F-AlF-NOTATATE was successfully prepared with a total synthesis time of 35 min. The optimized radiochemical yield was (23.8±3.1)% (without decay correction, n=3), the radioactivity was (4.63±0.68) GBq, and the radiochemical purity was >95%. The stability was good, and the product quality met the requirements. 18F-AlF-NOTATATE showed clear imaging in the patient with rectal segment NET, with SUV max of 13.3 and tumor/liver ratio of 3.3. Metastatic lesions in the liver, lymph nodes, and ribs showed high SUV max and tumor/liver ratios. The imaging of the pancreatic NET patient showed an abnormal increase in local radioactive uptake at the uncinate process of the pancreatic head, with SUV max of 5.6 and SUV max of 6.3 and the tumor/liver ratio of 2.3 after 2-hours imaging. Conclusions:Using the GE-FASTLab2 synthesis module, 18F-AlF-NOTATATE can be prepared with high activity. The preparation is simple, the method is stable, and the product has high radiochemical purity. 18F-AlF-NOTATATE exhibits good imaging performance in NET patients, providing valuable information for diagnosis, treatment, and prognosis evaluation.

Objective:To evaluate the effect of extracorporeal shock wave on the phosphoproteomics of the spinal cord in rats with diabetic neuralgia.Methods:Thirty-six healthy male SPF-grade Sprague-Dawley rats, aged 2 months, weighing 200-250 g, were divided into 3 groups ( n=12 each) using the random number table method: control group (group C), diabetic neuralgia group (group D), and extracorporeal shock wave + diabetic neuralgia group (group E). The rats were continuously fed a common diet in group C, while the rats were fed a high-sugar and high-fat diet for 8 weeks in D and E groups. Streptozotocin 35 mg/kg was intraperitoneally injected, and the successful induction of diabetic neuralgia was defined as the blood glucose >14.6 mmol/L and the mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) ≤85% of baseline values. Group E received extracorporeal shock wave treatment after developing the model, with 1, 000 shocks per session at a frequency of 10 Hz and an energy of 1.0 bar, once per week for a total of 4 sessions. The MWT and TWL were measured before developing the model (T 0) and at 1, 2, 3 and 4 weeks after developing the model (T 1-T 4). After the last extracorporeal shock wave treatment, the rats were anesthetized and sacrificed, and lumbar spinal cord tissues were obtained for proteomic analysis and for detection of the expression of glial fibrillary acidic protein (GFAP), interleukin-1beta (IL-1β), and tumor necrosis factor-alpha (TNF-α) (by immunohistochemistry). Results:Compared with group C, the MWT and TWL were significantly decreased at T 1-T 4 in D and E groups ( P<0.05). Compared with group D, the MWT and TWL were significantly increased at T 1-T 4 in group E ( P<0.05). The results of phosphoproteomics screening revealed 284 differentially phosphorylated proteins in D and C groups, 282 in E and C groups, and 303 in E and D groups ( P<0.05). The results of immunohistochemistry showed that the expression of GFAP, IL-1β and TNF-α was significantly up-regulated in group D compared with group C ( P<0.05); the expression of GFAP, IL-1β and TNF-α was significantly down-regulated in group E compared with group D ( P<0.05). Conclusions:The mechanism by which extracorporeal shock wave alleviates diabetic neuralgia is related to inhibition of astrocyte activation and excessive phosphorylation of mGluR5 in rats.