OBJECTIVE To study the sedative and hypnotic effects of zolpidem and the content of amino acid neurotransmitters in the thalamus and hypothalamus after treatment with zolpidem.METHODS Experiments on the loss of righting reflex(LORR)induced by the upper-threshold dose pentobarbital sodium(50 mg·kg-1,ip)were conducted to establish a hypoxic insomnia model in mice by simulating an altitude of 5500 m.Based on this model,the synergistic effect of zolpidem(0.33,1,3,9 and 27 mg·kg-1,ip)and the subthreshold(20 mg·kg-1,ip)and upper-threshold pentobarbital sodium,as well as the sedative hypnotic effect of zolpidem(10,13,17,20,23,30 and 40 mg·kg-1,ip)were evaluated via the LORR in normoxic and hypoxic environments.One hour after ip given zolpidem,the levels of glutamic acid(Glu)and γ-aminobutyric acid(GABA)in the thalamus and hypothalamus of mice in either environment were determined by the high-performance liquid chromatography(HPLC)with fluorescence detection.RESULTS One-day treatment with hypoxia significantly shortened the duration of LORR induced by the upper-threshold dose pentobarbital sodium.Compared with normoxia vehicle and hypoxia induced insomnia vehicle groups,zolpidem 9 and 27 mg·kg-1 significantly shortened the latency to LORR(P<0.01,P<0.05)and prolonged duration of LORR induced by subthreshold and upper-threshold pentobarbital sodi-um(P<0.01,P<0.05).The median effective dose(ED50)of LORR induced by zolpidem was 16.21 and 20.55 mg·kg-1 in normoxic and hypoxic environments,respectively.The results of neurotransmitter level detection showed that Glu contents in the thalamus and hypothalamus and the ratio of Glu/GABA in the hypothalamus were decreased after treatment with zolpidem 40 mg·kg-1 in a normoxic environment(P<0.01,P<0.05).Compared with the normoxia control group,Glu content and the ratio of Glu/GABA in the hypothalamus were significantly increased after treatment with hypoxia(P<0.01,P<0.05),and zolpidem 40 mg·kg-1 could reverse their elevation.CONCLUSION The sedative-hypnotic effect of zolpidem is weakened in a hypoxic environment,and the effect of zolpidem on the levels of Glu and GABA in the hypothalamus may play an important role in the sedative-hypnotic effect of zolpidem.

Objective To investigate the clinical efficacy of Kangfuxin solution combined with lidocaine and gentamicin in the treatment of oral ulcers. Method A total of 120 patients with oral ulcer admitted to Tongzhou District Maternity and Child Health Care Hospital of Beijing from May 2019 to July 2021 were selected and divided into observation group and control group randomly. 58 patients were treated with lidocaine as a control group and 62 patients in the observation group were treated with Kangfuxin solution combined with lidocaine and gentamicin. The clinical efficacy, including oral ulcer condition, visual analogue scale (VAS score) after 3d and 5d of treatment, inflammatory factor levels and adverse reactions were observed and compared between the two groups before and after treatment. Results The overall effective rate of the treatment group (95.12%) was higher than that of the control group (74.36%) and the difference was statistically significant (P<0.05). The ulcer area, ulcer healing time, and chewing improvement period were remarkably reduced compared with those in the control group (P<0.05). The VAS scores after 3d and 5d of treatment, the levels of tumor necrosis factor-α (TNF-α), C-reactive protein (CRP) and interleukin-6 (IL-6) between the two groups were statistically significant (P<0.05). There was no significant difference in the incidence of adverse reactions (3.22%) between the observation group and the control group (6.90%) (P>0.05). Conclusion Kangfuxin solution combined with lidocaine and gentamicin showed better curative effect on oral ulcer. This treatment regimen could effectively ameliorate various clinical symptoms of patients with oral ulcer, reduce patient pain and downregulate the levels of inflammatory factors, and help patients recover with high safety profile.

Gastric cancer is one of the malignancies with high incidence in the world. Xiangsha Liu Junzitang，a common prescription for the prevention and treatment of gastric cancer，has the effects of moving Qi to relieve pain，drying dampness， and invigorating the spleen. It is especially indicated for gastric cancer of the spleen and stomach qi deficiency syndrome. Based on the databases such as CNKI，Wanfang Data，and PubMed，the clinical efficacy and experimental studies of Xiangsha Liu Junzitang for the prevention and treatment of gastric cancer were summarized and sorted out，and the mechanism of Xiangsha Liu Junzitang for the prevention and treatment of gastric cancer was elaborated in order to provide useful references for the clinical and basic research on Xiangsha Liu Junzitang in the field of gastric cancer in the future. In clinical practice，Xiangsha Liu Junzitang can treat gastric precancerous lesions，increase the body immunity of patients with gastric cancer，improve the symptoms of spleen and stomach weakness after gastric cancer surgery，and reduce the adverse reactions of the digestive tract after chemotherapy for gastric cancer. Its clinical efficacy is superior to that of western medicine alone whether it is combined with western medicine or used alone. In the experimental research，Xiangsha Liu Junzitang has the effects of regulating inflammatory factors，inhibiting the proliferation of gastric cancer cells，promoting the apoptosis of gastric cancer cells，and improving the activity of pepsin. Modern pharmacological research has shown that Xiangsha Liu Junzitang can conduct a comprehensive intervention with multiple components and multiple targets. The main components of a single drug contained include saponins，polysaccharides，lactones，volatile oils，organic acids，and others， with the effects of protecting gastric mucosa，regulating endocrine，and promoting apoptosis of epithelial cells in gastric mucosal dysplasia，reflecting the advantages and values of traditional Chinese medicine in the prevention and treatment of gastric cancer.

Objective To investigate the infection of human parvovirus B19 in Suzhou voluntary blood donors under the current blood screening model. Methods A total of 893 blood donor samples from September to December 2022 were randomly collected. Samples were tested to determine the seroprevalence (anti-B19 IgG and IgM) of B19 antibodies by enzyme-linked immunosorbent assay(ELISA), and B19 DNA of positive samples was further detected by real-time polymerase chain reaction(PCR) assay. Results Among 893 samples, the total seroprevalence of B19 antibody was 20.7% (185/893), with anti-B19 IgG and IgM positive rate at 19.4% (173/893) and 1.9% (17/893), respectively, showing significant difference (P<0.05). No difference in the positive rates of B19 IgG and IgM (20.1%, 1.5% vs 18.0%,2.6%) was noticed by gender(P>0.05). The prevalence of anti-B19 IgG statistically increased with age (P<0.05), while there was no difference in the prevalence of anti-B19 IgM (P>0.05). No statistical difference was not found in anti-B19 IgG and IgM samples among different blood groups. The anti-B19 IgG in repeated blood donors was higher than that in first-time donors(21.5% vs 15.9%)(P<0.05) while there was no difference in the positive rate of IgM antibodies (P>0.05). Three cases were found to be positive for B19 DNA in the B19 antibody positive samples, with the positive rate at 1.6%(3/185). Conclusion Although the prevalence of B19 infection in Suzhou was lower than that in other areas and was mostly past infection, there was still a certain proportion of persistent infection and acute infection, which posed the potential risk of blood transfusion transmission. Therefore, attention should be paid to blood transfusions, especially for the high-risk and susceptible groups.

ZHANG Zhongjing's Zhenwutang is a classic formula for warming Yang and excreting water. It is composed of Aconiti Lateralis Radix Praeparata， Poria， Atractylodis Macrocephalae Rhizoma， Paeoniae Radix Alba， and Zingiberis Rhizoma Recens. Physicians of later generations have inherited and developed this formula by detailed recording and application. This paper adopted bibliometrics method to analyze Zhenwutang in terms of history， indications， dosage， drug processing， usage， and modification. The results showed that Zhenwutang was most widely used in Ming and Qing dynasties. Many physicians have inherited ZHANG Zhongjing's theory regarding the application of Zhenwutang in disease treatment， and a few physicians have used it to treat diphtheria and water-related diseases. Some physicians modified this formula to treat maculae， intermittent dysentery， jaundice and so on. Zhenwutang was mainly used to treat diseases of the circulatory system， respiratory system and urinary system in modern clinical practice. The processing of herbal medicines in this formula was clear. Specifically， the raw material of Aconiti Lateralis Radix Praeparata needed to be processed and peeled， while those of Poria， Atractylodis Macrocephalae Rhizoma， Paeoniae Radix Alba， and Zingiberis Rhizoma Recens can be used directly. Although being different， most of the dosages were consistent with those in Treatise on Febrile Diseases. According to the textual research， it is suggested that the reference dosage of this prescription in clinical practice is 41.25 g for Poria， Paeoniae Radix Alba， and Zingiberis Rhizoma Recens， respectively， 27.5 g for Atractylodes macrocephala， and 15 g for Aconiti Lateralis Radix Praeparata. The medicinal materials should be decocted in 1 600 mL water to reach a volume of 600 mL. After removal of the residues， the decoction should be taken warm with 140 mL each time， three times a day. The textual research of Zhenwutang is expected to provide a theoretical reference for the clinical application and formulation of Zhenwutang.

ObjectiveTo study the changes of primary metabolites and phenols in the fruits of Acanthopanax senticosus at different development stages, so as to provide a theoretical basis for the rational utilization of A. senticosus fruit resources. MethodThe primary metabolites and phenols in the fruits at different development stages were determined via gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) and then compared by multivariate statistical analysis. ResultA total of 274 chromatographic peaks were obtained by GC-MS-based non-targeted metabonomics and 24 differential metabolites were screened out by multivariate statistical analysis. The differential metabolites were mainly concentrated in pentose phosphate pathway, galactose metabolism, ascorbic acid and aldose metabolism pathways. After color conversion, the pentose phosphate pathway and galactose metabolism were activated and increasing sugars were accumulated. The ascorbic acid and aldose metabolism pathways were active before color conversion, with high accumulation of the end product ascorbic acid. The ultra-high liquid chromatography-mass spectrometry (UPLC-MS) identified 28 phenols in the fruits at different development stages. Flavonoids were accumulated mainly at the green ripening stage before color conversion, and phenolic acids were accumulated mainly after color conversion. ConclusionThe accumulation of primary metabolites and phenols in A. senticosus fruits varies significantly among different development stages

BACKGROUND: Pulmonary microvascular endothelial cells (PMVECs) were not complex, and the endothelial barrier was destroyed in the pathogenesis progress of acute lung injury (ALI)/acute respiratory distress syndrome (ARDS). Previous studies have demonstrated that hepatocyte growth factor (HGF), which was secreted by bone marrow mesenchymal stem cells, could decrease endothelial apoptosis. We investigated whether mTOR/STAT3 signaling acted in HGF protective effects against oxidative stress and mitochondria-dependent apoptosis in lipopolysaccharide (LPS)-induced endothelial barrier dysfunction and ALI mice. METHODS: In our current study, we introduced LPS-induced PMEVCs with HGF treatment. To investigate the effects of mammalian target of rapamycin (mTOR)/signal transducer and activator of transcription 3 (STAT3) pathway in endothelial oxidative stress and mitochondria-dependent apoptosis, mTOR inhibitor rapamycin and STAT3 inhibitor S3I-201 were, respectively, used to inhibit mTOR/STAT3 signaling. Moreover, lentivirus vector-mediated mTORC1 (Raptor) and mTORC2 (Rictor) gene knockdown modifications were introduced to evaluate mTORC1 and mTORC1 pathways. Calcium measurement, reactive oxygen species (ROS) production, mitochondrial membrane potential and protein, cell proliferation, apoptosis, and endothelial junction protein were detected to evaluate HGF effects. Moreover, we used the ALI mouse model to observe the mitochondria pathological changes with an electron microscope in vivo. RESULTS: Our study demonstrated that HGF protected the endothelium via the suppression of ROS production and intracellular calcium uptake, which lead to increased mitochondrial membrane potential (JC-1 and mitochondria tracker green detection) and specific proteins (complex I), raised anti-apoptosis Messenger Ribonucleic Acid level (B-cell lymphoma 2 and Bcl-xL), and increased endothelial junction proteins (VE-cadherin and occludin). Reversely, mTOR inhibitor rapamycin and STAT3 inhibitor S3I-201 could raise oxidative stress and mitochondria-dependent apoptosis even with HGF treatment in LPS-induced endothelial cells. Similarly, mTORC1 as well as mTORC2 have the same protective effects in mitochondria damage and apoptosis. In in vivo experiments of ALI mouse, HGF also increased mitochondria structural integrity via the mTOR/STAT3 pathway. CONCLUSION In all, these reveal that mTOR/STAT3 signaling mediates the HGF suppression effects to oxidative level, mitochondria-dependent apoptosis, and endothelial junction protein in ARDS, contributing to the pulmonary endothelial survival and barrier integrity.
Animals ; Apoptosis ; Calcium/metabolism* ; Endothelial Cells/metabolism* ; Endothelium/metabolism* ; Hepatocyte Growth Factor/metabolism* ; Lipopolysaccharides/pharmacology* ; Mammals/metabolism* ; Mechanistic Target of Rapamycin Complex 1/metabolism* ; Mechanistic Target of Rapamycin Complex 2/metabolism* ; Mice ; Mitochondria/metabolism* ; Oxidative Stress ; Reactive Oxygen Species/metabolism* ; Respiratory Distress Syndrome ; Sirolimus/pharmacology* ; TOR Serine-Threonine Kinases/metabolism*

Targeted programmed death-ligand 1 (PD-L1) and CXC chemokine receptor type 4 (CXCR4), gene sequences encoding anti-PD-L1 nanobody and anti-CXCR4 nanobody were cloned into the pET-22b (+) vector to construct recombinant expression plasmid of anti-PD-L1&CXCR4 bispecific nanobody, which was connected with 6 × His tag and transformed into E.coli BL21 (DE3). The expressed proteins were then found to exist as a soluble form in the supernatant of bacterial lysate after induction of IPTG.Three purification methods were used to obtain the target protein in order to improve the yield and purity of the bispecific nanobody.The bacterial supernatant was separated and purified by His Trap FF affinity chromatographic column.The target protein output could exceed 1 mg/L, and the product purity could reach up to 97%.Besides, the anti-PD-L1&CXCR4 bispecific nanobody shows a specific binding ability to two antigens on the cell surface, enhancing the cytotoxicity of IL-2 activated human peripheral blood mononuclear cells (PBMC) to tumor cell line AsPC-1, which lays the foundation for further evaluation of its drug efficacy in vivo.

Objective:To investigate clinical manifestations and dermoscopic characteristics of lichen planus-like keratosis (LPLK) .Methods:Clinical data were collected from 21 patients with LPLK who visited Shanghai Skin Disease Hospital and underwent both dermoscopic and histopathological examinations from January 2017 to September 2019, and clinical and dermoscopic features were retrospectively analyzed.Results:These patients were aged 64.69 ± 13.29 years, and the ratio of males to females was 1∶2. Skin lesions were located on the face of 18 cases and legs of 3 cases, and were red/violaceous in color in 7 cases, reddish-brown in 5, brown/gray in 8, and brown/reddish in 1. There were 3 types of skin lesions, including plaque-like type in 10 cases, flat pigmented patch type in 6, and flat erythema-like type in 5. As dermoscopy showed, 12 cases were non-pigmented LPLK, and 9 were pigmented LPLK. Pigment granules were found in 13 lesions, and there was no significant difference in the prevalence of pigment granules between pigmented and non-pigmented LPLK ( P=0.07) ; pigment granules were often diffusely distributed (9/13) , and the diffuse distribution pattern was common paticularly in pigmented LPLK (8/9) ; locally distributed pigment granules were found in 4 cases of non-pigmented LPLK. Coarse pigment granules were seen in 10 cases (10/13) , including 8 of pigmented LPLK and 2 of non-pigmented LPLK, and the prevalence rate of coarse pigment granules significantly differed between the pigmented LPLK and non-pigmented LPLK groups ( P=0.002) . Moreover, special distribution patterns of pigment granules included the annular granular pattern (8/13) and peppered pattern (7/13) , and no significant difference was observed in the prevalence of the 2 special distribution patterns between the pigmented LPLK and non-pigmented LPLK groups (both P > 0.05) . Scales were seen in 13 cases (13/21) , and vascular structures in 7 (7/21) , and there was no significant difference in the prevalence of the 2 structures between the pigmented and non-pigmented LPLK groups ( P=0.67, 0.16, respectively) . Conclusions:LPLK mostly occurs on the face, and manifests as solitary red, reddish-brown or brownish-gray plaques or patches, whose surfaces may be covered with scales. The characteristic dermoscopic feature of LPLK is the presence of pigment granules, which are coarse, often diffusely distributed, and commonly observed in pigmented LPLK.

Objective:To analyze clinical and histopathological features of cutaneous horns, and to explore their relationship.Methods:A total of 204 patients with cutaneous horns confirmed by clinical and histopathological examinations were collected from Department of Pathology, Shanghai Skin Disease Hospital between January 2014 and May 2020, and their clinical and histopathological data were retrospectively analyzed.Results:Among 204 patients, there were 88 males and 116 females with the age being 70.47±14.76 years and median ( P25, P75) course being 6 (6, 24) months. Cutaneous horns were solitary in 201 cases, and multiple in 3. There were a total of 207 cutaneous horns, including 163 (78.74%) located on the head, face and neck, 18 (8.70%) on the trunk and 26 (12.56%) on the extremities. According to histopathological conditions at the base of the cutaneous horns, 154 (74.40%) cutaneous horns were considered to arise from benign lesions (named as benign horns) , including viral warts ( n=129) , seborrheic keratosis ( n=15) , inverted follicular keratosis ( n=3) , trichilemmomas ( n=5) , pyogenic granuloma ( n=1) and calcifying epithelioma ( n=1) ; 40 (19.32%) were considered to arise from premalignant lesions (actinic keratosis) and named as premalignant horns; 13 (6.28%) were considered to arise from malignant lesions (named as malignant horns) , including Bowen′s disease ( n=5) and squamous cell carcinoma ( n=8) . The average ages of patients with premalignant and malignant horns were significantly higher than those of patients with benign horns ( H=4.05, 3.16, respectively, both P < 0.01) . The base width of the malignant horns was significantly higher than that of the premalignant and benign horns ( H=2.74, 3.49, P < 0.05, 0.01, respectively) , and there was no significant difference between the premalignant and benign horns ( H=0.73, P > 0.05) . The height to base width ratio was significantly lower in the premalignant and malignant horns than in the benign horns ( H=4.06, 3.72, respectively, both P < 0.01) . The incidence rate of basal erythema was significantly higher in the premalignant and malignant horns than that in the benign horns ( χ2=29.57, 6.23, P < 0.01, 0.05, respectively) , and bleeding and crusting occurred more frequently in the malignant horns compared with the benign horns ( χ2=4.89, P < 0.05) . Conclusions:The base of most cutaneous horns is benign, which mainly includes viral warts, and premalignant and malignant horns account for about a quarter. Male, older age, wide base, low ratio of the height to base width, basal erythema, history of bleeding and crusting can serve as clues to early recognition of malignant lesions at the base of cutaneous horns.