ObjectiveTo establish a quality evaluation method for Zhuye Shigao granules（Zhuye Shigaotang） based on fingerprint and determination of index components， and to investigate the therapeutic effect of Zhuye Shigao granules on mice with cold-dampness pestilence attacking lung syndrome. MethodsThe fingerprint of Zhuye Shigao granules was established by high performance liquid chromatography（HPLC）， and the methods for determination of total calcium， orientin， isoorientin， ginsenosides Rg₁， Re and Rb₁ and other 2 index components were established. Fifty ICR mice were randomly divided into the blank group， model group， Zhuye Shigao granules low， medium and high dose groups（9.3， 18.6， 37.2 g·kg^-1·d^-1）， with 10 mice in each group. In addition to the blank group， the model mice with cold-dampness pestilence attacking lung syndrome was prepared by nasal drip of lipopolysaccharide combined with cold-dampness environment. Each administration group was given the corresponding liquid by gavage according to the dose， while the blank group and model group were given the same volume of normal saline by gavage. Then， the body temperature and organ index of mice in each group were measured， hematoxylin-eosin（HE） staining was used to investigate the lung tissue injury of mice in each group， and enzyme-linked immunosorbent assay（ELISA） was used to detect the changes of tumor necrosis factor-α（TNF-α）， interleukin（IL）-lβ， IL-6， IL-10 levels in serum and lung tissue， as well as immunoglobulin（Ig）A and IgM levels in serum. ResultsThe fingerprint similarity of 10 batches of Zhuye Shigao granules was>0.950， and 20 common peaks were calibrated. Seven of them were identified， including peak 11（isoorientin）， peak 12（orientin）， peak 14（apioside liquiritin）， peak 15（liquiritin）， peak 17（apioside isoliquiritin）， peak 19（isoliquiritin） and peak 20（liquiritigenin）. The results of quantitative analysis showed that the content range of each index component in 10 batches of Zhuye Shigao granules was as follows：Total calcium of 9.978-11.294 mg·g^-1， isoorientin of 0.033-0.041 mg·g^-1， orientin of 0.046-0.055 mg·g^-1， ginsenoside Rg₁+ginsenoside Re of 0.748-0.762 mg·g^-1， ginsenoside Rb₁ of 0.151-0.197 mg·g^-1， liquiritin of 1.106-1.366 mg·g^-1， glycyrrhizic acid of 0.904-1.182 mg·g^-1_. Compared with the blank group， the body temperature of mice in the model group was significantly increased， the organ indexes of liver， lung and spleen were significantly decreased， the organ index of thymus was significantly increased， HE staining of lung tissue showed infiltration of inflammatory cells， a small amount of serous exudation was observed in the alveoli， and lung tissue was damaged. After the intervention of Zhuye Shigao granules， the pathological changes were improved compared with the model group. The expression levels of IL-1β， IL-6 and TNF-α were significantly increased， the expression level of IL-10 was significantly decreased in serum and lung tissue. The levels of IgA and IgM in serum were significantly decreased（P<0.01）. Compared with the model group， the body temperature， the organ indexes and immune factor levels in serum and lung tissue of mice in the Zhuye Shigao granules medium and high dose groups were significantly reduced（P<0.05， P<0.01）. ConclusionIn this study， the quality evaluation of Zhuye Shigao granules was carried out based on fingerprint combined with determination of index components， and the fingerprint of four herbs（Lophatheri Herba， Ophiopogonis Radix， Pinelliae Rhizoma and Glycyrrhizae Radix et Rhizoma） in this formula and the determination of 8 index components were established. The therapeutic effect of Zhuye Shigao granules on mice with cold-dampness pestilence attacking lung syndrome may be related to inhibiting inflammatory response and mediating immune regulation.

Objective:To explore the predictive value of 18F-FDG PET/CT metabolic parameters combined with inflammatory markers for the medium-term efficacy of chemotherapy in patients with primary gastrointestinal diffuse large B cell lymphoma (PGI-DLBCL). Methods:From April 2011 to May 2020, 67 patients (37 males, 30 females, age: 28-85 years) with PGI-DLBCL examined by 18F-FDG PET/CT before chemotherapy in Changhai Hospital, Navy Medical University were retrospectively analyzed. All patients were treated with cyclophosphamide+ doxorubicin+ vincristine+ prednisone (CHOP) or rituximab+ CHOP (R-CHOP) regimens, and the medium-term efficacy was evaluated after 2-4 cycles of chemotherapy. The effect outcome was divided into complete remission (CR) group and non-CR (NCR) group based on the Lugano lymphoma response evaluation criteria. Mann-Whitney U test was used to compare the differences of SUV max, peak of SUV (SUV peak), metabolic tumor volume (MTV), total lesion glycolysis (TLG), platelet/lymphocyte ratio (PLR) and neutrophil/lymphocyte ratio (NLR) between two groups. The independent risk factors of NCR were analyzed by multivariate logistic regression and the binary logistic regression model was established according to the results. The model was tested with external validation data ( n=15). Results:Of 67 PGI-DLBCL patients, 28(41.8%) were CR and 39(58.2%) were NCR. SUV peak, MTV, TLG, PLR and NLR in NCR group (17.3(12.3, 28.1), 73.8(42.9, 141.7) cm 3, 887.5(300.9, 2 075.3) g, 203.9(155.7, 297.1), 3.9(3.0, 4.9)) were significantly higher than those in CR group (9.5(6.2, 15.2), 11.3(4.7, 23.2) cm 3, 85.2(35.5, 214.6) g, 149.3(102.8, 173.1), 2.2(1.8, 4.6); z values: from -6.41 to -2.33, all P<0.05). The logistic regression model was as follows: P=1/(1+ e - x), x=0.100×MTV+ 0.024×PLR-8.064. The prediction accuracy for NCR risk was 86.57%(58/67), with the accuracy of 13/15 tested by external validation data. Conclusion:MTV combined with PLR has a good predictive value for medium-term efficacy of CHOP/R-CHOP chemotherapy in patients with PGI-DLBCL.

Candida albicans is the most abundant fungal species in oral cavity. As a smart opportunistic pathogen, it increases the virulence by switching its forms from yeasts to hyphae and becomes the major pathogenic agent for oral candidiasis. However, the overuse of current clinical antifungals and lack of new types of drugs highlight the challenges in the antifungal treatments because of the drug resistance and side effects. Anti-virulence strategy is proved as a practical way to develop new types of anti-infective drugs. Here, seven artemisinins, including artemisinin, dihydroartemisinin, artemisinic acid, dihydroartemisinic acid, artesunate, artemether and arteether, were employed to target at the hyphal development, the most important virulence factor of C. albicans. Artemisinins failed to affect the growth, but significantly inhibited the hyphal development of C. albicans, including the clinical azole resistant isolates, and reduced their damage to oral epithelial cells, while arteether showed the strongest activities. The transcriptome suggested that arteether could affect the energy metabolism of C. albicans. Seven artemisinins were then proved to significantly inhibit the productions of ATP and cAMP, while reduced the hyphal inhibition on RAS1 overexpression strain indicating that artemisinins regulated the Ras1-cAMP-Efg1 pathway to inhibit the hyphal development. Importantly, arteether significantly inhibited the fungal burden and infections with no systemic toxicity in the murine oropharyngeal candidiasis models in vivo caused by both fluconazole sensitive and resistant strains. Our results for the first time indicated that artemisinins can be potential antifungal compounds against C. albicans infections by targeting at its hyphal development.
Animals ; Mice ; Candida albicans ; Candidiasis, Oral/drug therapy* ; Antifungal Agents/pharmacology* ; Hyphae ; Artemisinins/pharmacology*

Isochrysis galbana is considered an ideal bait for functional foods and nutraceuticals of humans because of its high fucoxanthin(Fx)content.However,multi-omics analysis of the regula-tory networks for Fx biosynthesis in I.galbana has not been reported.In this study,we report a high-quality genome assembly of I.galbana LG007,which has a genome size of 92.73 Mb,with a contig N50 of 6.99 Mb and 14,900 protein-coding genes.Phylogenetic analysis confirmed the monophyly of Haptophyta,with I.galbana sister to Emiliania huxleyi and Chrysochromulina tobinii.Evolutionary analysis revealed an estimated divergence time between I.galbana and E.huxleyi of～133 million years ago.Gene family analysis indicated that lipid metabolism-related genes exhibited significant expansion,including IgPLMT,IgOAR1,and IgDEGS1.Metabolome analysis showed that the content of carotenoids in I.galbana cultured under green light for 7 days was higher than that under white light,and β-carotene was the main carotenoid,accounting for 79.09％of the total carotenoids.Comprehensive multi-omics analysis revealed that the content of β-carotene,antheraxanthin,zeaxanthin,and Fx was increased by green light induction,which was significantly correlated with the expression of IgMYB98,IgZDS,IgPDS,IgLHCX2,IgZEP,IgLCYb,and IgNSY.These findings contribute to the understanding of Fx biosynthesis and its regulation,pro-viding a valuable reference for food and pharmaceutical applications.

Background and Objectives: Psoriasis is a chronic inflammatory skin disease, which the mechanisms behind its initiation and development are related to many factors. DMSCs (dermal mesenchymal stem cells) represent an important member of the skin microenvironment and play an important role in the surrounding environment and in neighbouring cells, but they are also affected by the microenvironment. We studied the glucose metabolism of DMSCs in psoriasis patients and a control group to reveal the relationship among glucose metabolism, cell proliferation activity，and VEC (vascular endothelial cell) differentiation in vitro, we demonstrated the biological activity and molecular mechanisms of DMSCs in psoriasis. Methods: and Results: We found that the OCR of DMSCs in psoriatic lesions was higher than that in the control group, and mRNA of GLUT1 and HK2 were up-regulated compared with the control group. The proliferative activity of DMSCs in psoriasis was reduced at an early stage, and mRNA involved in proliferation, JUNB and FOS were expressed at lower levels than those in the control group. The number of blood vessels in psoriatic lesions was significantly higher than that in the control group (p＜0.05), which the mRNA of VEC differentiation, CXCL12, CXCR7, HEYL and RGS5 tended to be increased in psoriatic lesions compared to the control group, in addition to Notch3. Conclusions We speculated that DMSCs affected local psoriatic blood vessels through glucose metabolism, and the differentiation of VECs, which resulted in the pathophysiological process of psoriasis.

Objective:To evaluate the effect of electroacupuncture (EA) on pyroptosis in renal tubular epithelial cells of rats with acute kidney injury (AKI) induced by endotoxin.Methods:Twenty-four healthy clean-grade Sprague-Dawley rats of either gender, aged 6-8 weeks, weighing 160-182 g, were divided into 4 groups ( n=6 each) using a random number table method: control group (group C), group AKI, EA plus AKI group (group EA), sham EA at non-acupoint plus AKI group (group SEA). The model of endotoxemia was established by intraperitoneally injecting 10 mg/kg lipopolysaccharide.Bilateral 30 min EA stimulation of Zusanli and Shenyu (according to atlas of animal acupoint) was performed starting from 5 days before establishing the model (once a day) and at 30 min before lipopolysaccharide administration on the day of establishing the model, with disperse-dense waves, frequency of 15 Hz, and the needle was kept until 6 h after injection of LPS in group EA.EA was performed at the points 0.5 cm lateral to the acupoints of Zusanli and Shenyu in group SEA.At 6 h after LPS injection, blood was taken from the heart, and the concentrations of serum blood urea nitrogen (BUN) and creatinine (Cr) were detected by an automatic biochemical analyzer, and the serum concentrations of neutrophil gelatinase-associated lipocalin (NGAL), kidney injury molecule-1 (KIM-1), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) by enzyme-linked immunosorbent assay.The rats were then sacrificed, and the left renal cortex was obtained for determination of pyroptosis rate of renal tubular epithelial cells (by TUNEL). The right renal cortex was obtained to detect the expression of caspase-1 and IL-1β by Western blot, and the expression of caspase-1 mRNA and IL-1β mRNA was detected by real-time polymerase chain reaction. Results:Compared with group C, the concentrations of BUN, Cr, NGAL, KIM-1, TNF-α, and IL-6 were significantly increased, the pyroptosis rate of renal tubular epithelial cells was increased, the expression of caspase-1 and IL-1β protein and mRNA in the renal cortex was up-regulated in group AKI ( P<0.05). Compared with group AKI, the concentrations of BUN, Cr, NGAL, KIM-1, TNF-α, and IL-6 were significantly decreased, the pyroptosis rate of renal tubular epithelial cells was decreased, the expression of caspase-1 and IL-1β protein and mRNA in the renal cortex was down-regulated in group SEA ( P>0.05). Conclusion:The mechanism by which EA reduces AKI may be related to inhibiting pyroptosis in renal tubular epithelial cells of rats.

Objective To evaluate the effect of sufentanil on apoptosis in spinal cord neurons of mice with peripheral nerve injury. Methods One hundred and fifty clean-grade healthy male BALB∕c mice, aged 6-8 weeks, weighing 18-22 g, were divided into 3 groups ( n=50 each) using a random number table method: sham operation group (group Sham), peripheral nerve injury group (group PNI) and sufentanil group ( group SF) . The model of unilateral sciatic nerve injury was established in PNI and SF groups. After establishing the model, sufentanil 5. 0 μg∕kg was intraperitoneally injected once a day for 3 consecutive days in group SF, while the equal volume of normal saline was given instead of sufentanil in Sham and PNI groups. Five mice in each group were sacrificed at days 1, 3, 7, 14 and 28 after surgery ( T0-4 ) , and L4-6 segments of the injure ipsilateral spinal cord were removed for examination of pathological changes ( with a light microscope) and for determination of neuronal apoptosis ( by TUNEL assay) . The ap-optosis index ( AI) was calculated. Five mice in each group were sacrificed at T0-4 , and L4-6 segments of the injured ipsilateral spinal cord were removed for detection of the expression of Bcl-2, Bax and cleaved caspase-3 by Western blot. The ratio of Bcl-2 expression to Bax expression ( Bcl-2∕Bax ratio) was calculat-ed. Results Compared with group Sham, the AI was significantly increased, the expression of Bcl-2 pro-tein was down-regulated, and the expression of cleaved caspase-3 and Bax was up-regulated in PNI and SF groups ( P<0. 05) . Compared with group PNI, the AI was significantly decreased, the expression of Bcl-2 protein was up-regulated, the expression of cleaved caspase-3 and Bax was down-regulated, the Bcl-2∕Bax ratio was increased (P<0. 05), and the pathological changes were significantly attenuated in group SF. Conclusion Sufentanil can inhibit apoptosis in spinal cord neurons of mice with peripheral nerve injury.

Objective To investigate the role of circular RNA (circRNA) in the occurrence of psoriasis.Methods Mesenchymal stem cells were isolated from skin lesions of 15 patients with psoriasis (psoriasis group) and skin tissues of 15 healthy human controls (control group) separately,and then subjected to cultivation.Flow cytometry and cell differentiation assay were performed to identify these mesenchymal stem cells.RNA sequencing was conducted to measure the expression of circRNA,and detailed bioinformatics analysis was performed.Then,7 differentially expressed circRNAs were selected,and a circRNA-microRNA interaction network was established.In this network,3 related microRNAs were selected and validated by real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR).Statistical analysis was carried out by a two-sample t-test for comparison of qRT-PCR results between the psoriasis group and control group.Results As RNA sequencing showed,a total of 6 323 circRNAs were identified,and 3 227 out of these circRNAs were first reported in this study.Compared with the control group,129 circRNAs were differentially expressed in the psoriasis group,including 123 up-regulated circRNAs and 6 down-regulated circRNAs.The predicted circRNA-microRNA interaction network showed that several psoriasis-related microRNAs were associated with the 7 circRNAs,such as miR-17-5p,miR-30e-5p,miR-142-3p/5p,miR-369-3p,miR-184,miR-4490,miR-654-3p and miR-423-5p.qRT-PCR also confirmed that the 7 differentially expressed circRNAs were up-regulated in the psoriasis group.Compared with the control group,the expression of the 3 selected microRNAs significantly decreased in the psoriasis group (t =3.993,3.217,2.918,respectively,all P ＜ 0.05).Conclusion There is aberrant expression of circular RNAs in the mesenchymal stem cells from psoriatic skin lesions,which may take part in the occurrence of psoriasis.

Objective To investigate the value of spectral CT with iodine parameters in distinguishing moderately-differentiated adnocarcinoma from poorly-differentiated adenocarcinoma.Methods 61 patients with gastric adenocarcinoma underwent preoperative CT scanning that included arterial phase(AP) and venous phase(VP) with gemstone spectral imaging(GSI) mode.All measurements were performed on the GSI viewer.The iodine concentration (IC) and the water concentration (WC) of the primary lesion were measured.Then the normalized iodine concentration (NIC) and contrast enhancement ratio (CER) were calculated.The CT values were measured at 70 keV Mono image.All the values of CER, NIC, IC and WC in the primary lesion were recorded and assessed by independent-samples t test.Receiver operating characteristic (ROC) was used to determine the threshold of IC and NIC for differentiating poorly-differentiated adenocarcinoma and moderately-differentiated adnocarcinoma.Results The IC values between moderate differentiation group and poor differentiation group were 8.73±4.05 vs 11.07±4.80(100 μg/cm3) in AP and 16.89±4.89 vs 21.18±5.96(100 μg/cm3) in VP.The values of NIC between moderately differentiated group and poorly differentiated group were 0.10±0.06 vs 0.13±0.06 in AP and 0.38±0.10 vs 0.49±0.12 in VP respectively.Significant difference of NIC (tVP=3.38, PVP<0.01) and IC (tVP=2.87, PVP=0.01) were only found between poor differentiation group and moderate differentiation group in venous phase.The value of WC and CER (both P>0.05) in double phases revealed no significant difference between the two groups.Conclusion Iodine quantitative parameters of spectral CT can be potentially used to preoperatively evaluate the differentiation degree of gastric adenocarcinoma.Thus, the iodine parameters can reflect the differentiation degree of gastric adenocarcinoma.

Objective To evaluate the effect of sufentanil on the expression of basic fibroblast growth factor(bFGF)in nerve tissues after peripheral nerve injury in mice. Methods One hundred pathogen-free healthy male Balb∕c mice, aged 6-8 weeks, weighing 18-22 g, were divided into 4 groups (n=25 each)using a random number table: peripherial nerve injury group(group PNI), high dose sufentanil group(group H), medium dose sufentanil group(group M)and low dose sufentanil group (group L). The model of unilateral sciatic nerve transaction was established in ketamine-anesthetized mice. Sufentanil 100, 50 and 25 μg∕kg were intraperitoneally injected immediately after establishment of the model once a day for 3 consecutive days in H, M and L groups, respectively. The equal volume of normal saline was given instead in group PNI. On 1, 2, 4, 8 and 12 weeks after establishment of the model, 5 mice were sacrificed and the nerve tissues were obtained from the site 05 cm up and down the lesion site of the nerve for examination of the shape of the myelin sheath of nerve fibers(with an electronic microscope).The expression of bFGF in the sciatic nerve tissue was detected by Western blot. Results The shape of medulla sheath was irregular, the thickness of myelin lamellae was thin, the separation of myelin lamellae was aggravate, demyelinate was found, and the proliferation of Schwann cells was not marked in group PNI. The shape of medulla sheath was regular, the thickness of myelin lamellae was dense, and the proliferation of Schwann cells was marked in group H. The shape of medulla sheath was irregular, the separation of mye-lin lamellae was observed, demyelinate was found, and the proliferation of Schwann cells was not marked in group L. Compared with group PNI, the expression of bFGF was significantly up-regulated in H, M and L groups(P<005). Compared with group L, the expression of bFGF was significantly up-regulated in H and M groups(P<005). Compared with group M, the expression of bFGF was significantly up-regulated in group H(P<005). Conclusion The mechanism by which sufentanil improves regeneration and repair after peripheral nerve injury may be related to up-regulating the expression of bFGF in nerve tissues of mice.