Objective To evaluate the performance of microfluidic chips in the identification and genotyping of Malassezia species.Methods This study included 6 reference Malassezia strains and clinical Malassezia isolates from the scrapings of patients with pityriasis versicolor and follicular contents of patients with Malassezia folliculitis.These isolates were identified by DNA sequencing,random amplified polymorphic DNA (RAPD)-PCR and microfluidic chips.Cluster analysis was carried out and tree diagrams were generated.Results A total of 83 Malassezia isolates were obtained from 72 patients with pityriasis versicolor and 11 patients with Malassezia folliculitis.Genomic DNA of most strains was successfully amplified by PCR with two primers S22 and S24,and PCR with S22 primer produced more stable and clear amplification bands than that with S24.Positive bands of different sizes were repetitively obtained by using microfluidic chips,with interspecies and intraspecies polymorphisms observed in all the strains.On the basis of DNA sequencing,microfluidic chips and RAPD-PCR could be used to successfully distinguish the following eight species,i.e.,M.furfur,M.sympodialis,M.globosa,M.pacbydermatis,M.slooffiae,M.Japonica,M.yamatoensis and M.dermatis.Conclusions As a rapid,high-throughput and high-sensitivity method,microfluidic chips combined with RAPD-PCR shows an advantage for analyzing interspecies genetic diversity,genetic relationship of Malassezia species,as well as for identifying new Malassezia species.

AIM: To establish the method of determining peoniflorin,ferulic acid and icariin in Bushen Huoxue Granule(Herba Epimedii,Radix Angelicae Sinensis,Radix Paeoniae Alba,etc.).METHODS: HPLC-DAD was used to determine peoniflorin,ferulic acid and icariin.The samples were separated on Alltima C18(5 ?m,250 mm ? 4.6 mm) with mobile phase consisted of methanol-0.1% phosphoric acid(gradient elution).RESULTS: The average recoveries of peoniflorin,ferulic acid and icariin were 96.7%,97.5%,99.6% and RSD were 1.37%,3.13%,0.96%.respectively.The peoniflorin,ferulic acid and icariin samples showed a good relationship in the ranges of 0.318-3.970 ?g,0.096-1.200 ?g,0.414-5.175 ?g in the chosen HPLC condition.CONCLUSION: The method is simple,accurate and reproducible and can be used for the quality control of granule.

Aim To study the combined effect Euphor-bia kansui of and Glycyrrhiza uralensis on metabolism of Kansuinine A and Kansuinine B. Methods Control, GU and GU plus EK groups were treated for 10 days, respectively. Then the liver microsomes were pre-pared. KA and KB were incubated with microsomes of each group, and concentrations of KA and KB were measured to reflect the metablism of KA and KB. E-rythromycin, diphenhydramine hydrochloride,benzbro-marone, cimetidine, fluconazole the inhibitors of CYP3 A4 , CYP2 D6 , CYP2 C9 , CYP1 A2 , CYP2 C19 respectively, were incubated with KA and KB. Con-centrations of KA and KB were measured to reveal the cytochrome P450 isoforms which involved in the metab-olism of them. KA and KB were incubated with glycyr-rhizic acid and enoxolone. Then concentrations of KA and KB were measured to reveal the effects of glycyr-rhizic acid and enoxolone to KA and KB. Results Concentrations of KA and KB in GU plus EK group were significantly higher than those in control and EK groups, which showed that the metablisom of KA and KB was inhibited in GU plus EK group. In addition, concentrations of KA and KB were higher in microsomes incubated with erythromycin, diphenhydramine hydrochloride, benzbromarone, cimetidine and fluconazole than in control group, which revealed that the metablism of KA and KB was slowed down when CYP3A4, CYP2D6, CYP2C9, CYP1A2 and CYP2C19 were inhibited. They may participate in the metablisom of KA and KB. After incubation with glycyrrhizic acid and enoxolone, the metablism of KA and KB was slowed down. Conclusions KA and KB may be the substrates of CYP2C19. GU plus EK can inhibit the activity of CYP2C19, which resulted in KA and KB metablism slowing down and accumulation. In addition, glycyrrhizic acid and enoxolone can inhibit the metablism of KA and KB. It may be one of the reasons of increased toxicity of GU plus EK.

Objective To explore the clinical characteristics of metabolic disorder and the incidence rate of metabolic syndrome (MS) in the patients with posthepatitic cirrhosis.Methods Four hundred and fifty-five patients with posthepatitic cirrhosis were included in the study and divided into hepatitis B related cirrhosis group (LCB group,351 cases) and hepatitis C related cirrhosis group (LCC group,104cases).The prevalence of metabolic disorder was recorded and the characteristics of posthepatitic cirrhosis with MS and without MS were compared and analysed.Results The prevalence of hyperglycemia,hypertension,hyperlipemia,obesity and MS in the posthepatitic cirrhosis patients was 46.59% (212/455),15.16% (69/455),15.38% (70/455),22.64% ( 103/455),12.53% (57/455) respectively.The prevalence of MS in LCB and LCC was 8.26% (29/351) and 26.92% (28/104).The levels of body mass index (BMI),fasting blood glucose,hypertension,cholesterol,aminotransferase (ALT) in LCB with MS patients were significantly higher than those in LCB without MS patients.There were no differences in the levels of HBeAg and HBV DNA between LCB with MS patients and LCB without MS patients.The levels of BMI,hypertension,triglyceride in LCC with MS patients were significantly higher than those in LCC without MS patients.There were no differences in fasting blood glucose,cholesterol and ALT between LCC with MS patients and LCC without MS patients.Logistic regression revealed that BMI was the independent factor in LCB and LCC with MS.Conclusions The prevalence of hyperglycemia and obesity are higher in LCB and LCC.The incidence rate of MS in LCB is less than that in the general population,while the incidence rate of MS in LCC is significantly higher than that in the general population,and it's nothing to do with the viral replication.BMI is an important factor affected in posthepatitic cirrhosis with MS.

AIM: To study the effect of Qingqi Liangying Injection on febrile rats induced by zymonsa and on content of PGE_2 as well as cAMP in rat hypothalamus. METHODS: The rat febrific model induced by 10% zymonsa solution was used to observe QingqiLiangying Injection's effect on the rat temperature, and the radioimmunoassay(RIA) was used to determin the content of PGE_2 and cAMP in hypothalamus of rats. RESULTS: Qingqi Liangying Injection had an obvious antipyretic effect on febrile rats; Qingqi Liangying Injection could significantly reduce the content of PGE_2 and cAMP. CONCLUSION: Qingqi Liangying Injection can obviously reduce the rat body temperature and the content of PGE_2 and cAMP in hypothalamus of rats.

Objective:To investigate high mobility group protein 1(HMGB1), tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)levels and their clinical significance in elderly patients with viral pneumonia.Methods:One hundred and sixty elderly patients with viral pneumonia admitted to the Sixth Hospital Affiliated to Anhui Medical University were enrolled as research subjects.In addition, 40 elderly people who underwent regular physical examination were considered as the control group.Patients with viral pneumonia were divided into the low-risk group, middle-risk group and high-risk group according to CURB-65 scores and pneumonia severity index(PSI)scores.HMGB1, TNF-α and IL-6 levels were compared between different groups.The correlations of CURB-65 scores and PSI scores with HMGB1, TNF-α, IL-6 levels were analyzed.Multivariate Logistic regression analysis was used to examine influencing factors for the severity of viral pneumonia in elderly patients.Results:HMGB1, TNF-α and IL-6 levels were higher in research subjects than in the control group.As the severity of viral pneumonia increased, so did HMGB1, TNF-α and IL-6 levels(all P<0.05). HMGB1, TNF-α and IL-6 levels in the severe viral pneumonia group were significantly higher than those in the non-severe viral pneumonia group( P<0.05). HMGB1, TNF-α and IL-6 levels were positively correlated with CURB-65 scores and PSI scores(CURB-65 score: r=0.463, 0.392 and 0.497, P=0.015, 0.003 and 0.025; PSI score: r=0.596, 0.515 and 0.381, P=0.007, 0.011 and 0.009). HMGB1, TNF-α and IL-6 levels were influencing factors for the severity of viral pneumonia in elderly patients( OR=1.344, 1.422 and 1.351, P=0.006, 0.015 and 0.009). Conclusions:HMGB1, TNF-α and IL-6 levels are closely correlated with the severity of viral pneumonia and are helpful for early assessment of viral pneumonia.

Objective To observe the effect of combined opioid-free and loco-regional anesthesia on the quality of recovery after retrolaparoscopic renal surgery.Methods Sixty patients undergoing elective retrolaparoscopic renal surgery were divided into two groups(30 patients per group):the opioid-free anesthesia group(OFA group)and conventional anesthesia group with opioid(OA group)u-sing the random number table method.Both groups underwent ultrasound-guided anterior quadratus lumbar block(QLB)30minutes be-fore induction of anesthesia.In the OFA group,dexmedetomidine was given before induction,and induction was performed with propofol,midazolam,esketamine,and rocuronium,maintained with dexmedetomidine,propofol,sevoflurane anesthesia,and intermittent push of esketamine.In the OA group,propofol,midazolam,sufentanil,and rocuronium were given to induce anesthesia and maintained with remifentanil,propofol,and sevoflurane.Both groups were given intravenous self-administered analgesia after surgery.The mean arterial pressure(MAP)and heart rate(HR)at different time points,and the resting and motor VAS scores at 2,6,12,24 and 48h after sur-gery were recorded.QoR-40 was used to compare the quality of recovery at 24 hours after surgery between the two groups,and adverse effects and recovery of patients were recorded.Results QoR-40 scores were higher in the OFA group than that in the OA group(P＜0.05).Postoperative pain,postoperative opioid consumption,and incidence of nausea and vomiting were reduced in the OFA group(P＜0.05);and time to first ambulation,first exhaustion,and length of hospital stay were all reduced(P＜0.05).Conclusion Opioid-free anesthesia combined with quadratus lumbar block can significantly improve the quality of postoperative recovery,reduce postoperative pain,reduce postoperative opioid drug consumption,and promote postoperative recovery in patients undergoing retrolaparoscopic renal surgery.

Objective To investigate the effects of glucocorticoid combined with spleen aminopeptide on immune function and pulmonary function in children with combined allergic rhinitis and asthma syndrome (CARAS).Methods A total of 166 cases of CARAS were divided into observation group (84 cases) and control group (82 cases),the patients in the control group were treated by nasal inhalation of Budesonide Aerosol,in addition to the treatment of control group the observation group was given spleen aminopeptide oral lyophilized powder for treatment,and two groups were treated continuously for 3 months.The effect of 2 groups of children,and the changes of immune and lung function before and after treatment were compared.Results Rhinitis and asthma were significantly reduced in two groups of children after treatment,and the reductions of the score of rhinitis symptoms and the asthma score in the observation group were more significant than those in the control group(P＜0.05).The humoral immunity index (IgG,IgM and IgA) of the 2 groups increased significantly after treatment.CD3+,CD4+ and the ratio of CD4+/CD8+ in the cellular immune indexes increased significantly,and CD8+ decreased significantly.The immune indexes in the observation group were significantly improved compared with those in the control group (P＜0.05).After treatment,the forced expiratory volume in one second(FEV1) and maximal expiratory flow rate (PEFR) of the 2 groups increased significantly compared with those before treatment,and the degree of increase in the observation group was significantly higher than that in the control group(P＜0.01).Conclusion Glucocorticoid combined with spleen aminopeptide could not only improve the symptoms and signs of children with CARAS,but also significantly enhance cellular immunity,humoral immunity and pulmonary function.

OBJECTIVE To study the intervention effect and mechanism of Zhongfeng yure decoction on ischemic stroke model rats. METHODS Totally 85 rats were randomly divided into sham operation group （normal saline， n＝15）， model control group （normal saline， n＝18）， Nimodipine tablet group （positive control， 10.8 mg/kg， n＝18）， high-dose group of Zhongfeng yure decoction （20.52 g/kg， n＝17） and low-dose group of Zhongfeng yure decoction （5.13 g/kg， n＝17）， respectively. After 7 days of preventive continuous administration （once a day）， except for the sham operation group， the rats’ middle cerebral artery occlusion （MCAO） model was established by the modified suture method in other groups. After modeling， the rats in each group continued to be administered for 3 days. During experiment， general condition of the rats was observed， and the neurological function score was performed. After the last administration， the organ index was calculated， the cerebral infarction area and pathological changes of brain tissue were observed. The levels of tumor necrosis factor-α （TNF-α） and interleukin 6 （IL-6） in brain tissue and serum， and the average optical density value of caspase-3 and phosphorylated protein kinase B（p-AKT） protein in brain tissue were detected. RESULTS Three days after modeling， compared with sham operation group， the neurological function score， in brain tissue index， spleen tissue index， proportion of cerebral infarction area， the levels of TNF-α and IL-6 in brain tissue and serum， and the average optical density value of caspase-3 protein in brain tissue were significantly increased in the model control group （P＜0.05 or P＜0.01）； karyopyknosis， diffuse edema and other lesions appeared in brain tissue. Compared with the model control group， the above indexes in each administration group were improved to varying degrees. Among them， there were significant regression in brain tissue index， spleen tissue index， proportion of cerebral infarction area， TNF-α level in brain tissue and serum， and the average optical density values of caspase-3 protein and p-AKT protein in brain tissue of rats in high-dose group of Zhongfeng yure decoction （P＜0.05 or P＜0.01）. CONCLUSIONS Zhongfeng yure decoction has a certain intervention and therapeutic effect on MCAO model rats. The mechanism may be to reduce the secretion of inflammatory factors TNF-α and IL-6， down-regulate the expression of caspase-3 protein in ischemic brain tissue， up-regulate the expression of p-AKT protein， so as to protect the neurons.

ObjectiveTo evaluate the methodological quality of traditional Chinese medicine （TCM） diagnosis and treatment guidelines/consensus of constipation with Appraisal of Guidelines for Research and Evaluation Ⅱ （AGREE Ⅱ）tool， and to study the attention situation of the included Chinese patent medicines in China's National Reimbursement Drug List in the guidelines/consensus. MethodThe data of CNKI，VIP，Wanfang Data，SinoMed，PubMed and Cochrane from the inception of the databases to October 2021 were searched to collect the TCM diagnosis and treatment guidelines/consensus of constipation. Then，the diagnosis and treatment standards and recommended Chinese patent medicines were extracted. Two researchers assessed the methodological quality of the guidelines/consensus with AGREE Ⅱ tool independently. The quality of reports was evaluated by Reporting Items for practice Guidelines in HealThcare （RIGHT） Statement. The recommended Chinese patent medicines in the guidelines/consensus were compared with those in the National Reimbursement Drug List. ResultEleven consensus and 2 guidelines were included，involving 794 experts. The scores of AGREE II were clarity of presentation（59.0%），scope and purpose（44.0%），stakeholder involvement（23.1%），rigor of development （12.1%），applicability （11.1%），and editorial independence （8.3%） from high to low. Five articles were recommended at B level（recommended after revision） and 8 articles were at C level （not recommended）. The average scores of RIGHT Statement were as follows：basic information （93.59%），background （57.69%），evidence （18.46%），recommendations （20.88%），review and quality assurance （19.23%），funding，declaration and management of interests （0.00%）， and other information （0.00%）. The included guidelines/consensus recommended a total of 27 Chinese patent medicines，among which 20 were included in the National Reimbursement Drug List，with 4 species of Class A and 16 species of Class B， accounting for 74.1% of all recommended Chinese patent medicines. Ten purgative Chinese patent medicines in the National Reimbursement Drug List were recommended by the guidelines/consensus，accounting for 50% of all purgative drugs， and 8 were not recommended. There were prescriptions for purgation， for promoting digestion and removing food stagnation， for clearing heat and purging fire，and for warming the middle and dissipating cold，Tibetan medicine and Mongolian medicine. ConclusionBy the AGREE Ⅱ assessment，the methodological quality of the TCM diagnosis and treatment guidelines/consensus of constipation included in this study needed to be improved in the future. The report quality evaluated with RIGHT Statement was low. Most drugs included in the National Reimbursement Drug List were paid attention in the TCM diagnosis and treatment guidelines/consensus of constipation. Moreover，the drugs included in the National Reimbursement Drug List could basically fulfill the clinical needs reflexed from the guidelines/consensus recommendations. However， the reasons of some drugs failing to be included in the National Reimbursement Drug List needed to be studied in the future.