OBJECTIVE To study the improvement effects of Shaoyao gancao decoction （SGD） on acute lung injury （ALI） in rats and its effects on the intestinal flora. METHODS Sixty SD rats were randomly divided into normal group （CON group， normal saline）， model group （MOD group， normal saline）， positive control group （DEX group， 5 mg/kg dexamethasone）， SGD low-dose， medium-dose and high-dose groups （SGD-L， SGD-M， SGD-H groups， 5.8， 11.6， 23.2 g/kg， calculated by crude drug）， with 10 rats in each group. Each group was given relevant medicine 10 mL/kg intragastrically， for 7 consecutive days. Thirty minutes after the last administration， CON group was given constant volume of normal saline via airway infusion， and other groups were given lipopolysaccharide （5 mg/kg） via airway infusion to induce ALI model. After 12 hours of modeling， the lung tissue wet/dry weight ratio was calculated， and the contents of interleukin 1β （IL-1β）， IL-6 and tumor necrosis factor α（TNF-α） in rat bronchial alveolar lavage fluid （BALF） were all detected； the pathological changes of lung tissue were observed after hematoxylin-eosin staining. The intestinal flora of rat feces was analyzed by 16S rRNA sequencing technology， and the correlation of differential bacteria genera with inflammatory factors was also analyzed. RESULTS Compared with MOD group， the infiltration of inflammatory cells in the lung tissue of rats in each SGD dose group was decreased， and the thickening of alveolar septum and pulmonary edema improved； lung tissue wet/dry weight ratio， the levels of IL-1β， IL-6 and TNF-α in BALF significantly decreased （P＜0.05 or P＜0.01）. SGD （low dose） could improve the intestinal flora disorder in ALI rats， restore the diversity and richness of intestinal flora， regulate the structure of flora， reduce the abundance of Lactobacillus， Streptococcus and Escherichia-Shigella， and increase the abundance of Firmicutes， Lachnospira， Ruminococcus， Clostridia，Dubosiella and Akkermansia. Through correlation analysis， it was found that the relative abundance of Lactobacillus， Streptococcus and Escherichia-Shigella was positively related to the levels of inflammatory factor IL-1β， IL-6 and TNF-α （P＜0.05 or P＜0.01）. The relative abundance of Lachnospira， Dubosiella， Firmicutes was significantly negatively correlated with the levels of inflammatory factors mentioned above （P＜0.05 or P＜0.01）. CONCLUSIONS SGD may improve ALI by reducing lung tissue injury and inflammatory response and regulating flora structure in rats.

BACKGROUND: Women comprise more than half of people living with human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS) worldwide and incomplete immune recovery and metabolic abnormalities affect them deeply. Studies of HIV antiretroviral therapy (ART) have a low female representation in China. We aimed to investigate immune reconstitution and metabolic changes of female HIV-positive cohort in China longitudinally. METHODS: HIV-positive women who initiated ART from January 2005 to June 2021 and were followed up regularly at least once a year were included in this study. Immunological indicators (cluster of differentiation 4 [CD4] counts and CD8 counts), viral load (VL), and metabolic indicators were collected at follow-up. All data were collected from the China Disease Prevention and Control Information System (CDPCIS). VL was tested half a year, 1 year after receiving ART, and every other year subsequently according to local policy. CD4/CD8 ratio normalization was considered as the primary outcome and defined as a value ≥1. Incidence rate and probability of CD4/CD8 ratio normalization were estimated through per 100 person-years follow-up (PYFU) and Kaplan-Meier curve, respectively. Multivariate Cox regression was used to identify independent risk factors associated with CD4/CD8 ratio normalization. We further studied the rate of dyslipidemia, hyperuricemia, diabetes, liver injury, and renal injury after ART initiation with the chi-squared tests or Fisher's exact probability tests, and a generalized estimating equation model was used to analyze factors of dyslipidemia and hyperuricemia. RESULTS: A total of 494 female patients with HIV/AIDS started ART within 16 years from January 2005 to June 2021, out of which 301 women were enrolled with a median duration of ART for 4.1 years (interquartile range, 2.3-7.0 years). The overall incidence rate of CD4/CD8 ratio normalization was 8.9 (95% confidence interval [CI], 7.4-10.6) per 100 PYFU, and probabilities of CD4/CD8 normalization after initiating ART at 1 year, 2 years, 5 years, and 10 years follow-up were 11.7%, 23.2%, 44.0%, and 59.0%, respectively. Independent risk factors associated with CD4/CD8 normalization were baseline CD4 cell counts <200 cells/μL, CD8 counts >1000 cells/μL, and more than 6 months from the start of combined ART (cART) to first virological suppression. Longitudinally, the rate of hypercholesterolemia (total cholesterol [TC]) and high triglyceride (TG) showed an increasing trend, while the rate of low high-density lipoprotein cholesterol (HDL) showed a decreasing trend. The rate of hyperuricemia presented a downtrend at follow-up. Although liver and renal injury and diabetes persisted during ART, the rate was not statistically significant. Older age and protease inhibitors were independent risk factors for increase of TC and TG, and ART duration was an independent factor for elevation of TC and recovery of HDL-C. CONCLUSIONS This study showed that women were more likely to normalize CD4/CD8 ratio in comparison with findings reported in the literature even though immune reconstruction was incomplete.
Humans ; Female ; CD4-CD8 Ratio ; HIV ; Immune Reconstitution ; Hyperuricemia/drug therapy* ; HIV Infections/drug therapy* ; Acquired Immunodeficiency Syndrome/drug therapy* ; Anti-Retroviral Agents/therapeutic use* ; Cholesterol ; Viral Load ; CD4 Lymphocyte Count ; Anti-HIV Agents/therapeutic use*

ObjectiveTo study the effect of Huangqintang on intestinal T helper cells（Th）17/regulatory T cells（Treg） and Th1/Th2 balance in experimental ulcerative colitis （UC） mice. MethodSixty male C57BL/6J mice were randomly divided into normal group， model group， sulfasalazine group （0.2 g·kg^-1）， and Huangqintang low-dose， medium-dose and high-dose groups （4.55，9.1，18.2 g·kg^-1， respectively） according to body weight. Except for the normal group， all mice were given 2.5% dextran sodium sulfate （DSS， added to free drinking water） for 5 days to induce UC. After 7 days of continuous intragastric administration， the mice were sacrificed and the mesenteric lymph nodes （MLNs） and colon tissues were collected. The Th17/Treg and Th1/Th2 cell ratios in MLNs of mice were measured by flow cytometry. Interleukin-2 （IL-2）， interleukin-17A （IL-17A）， interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， interferon-γ （IFN-γ）， interleukin-4 （IL-4）， interleukin-10 （IL-10） and other cytokines in colon tissues were detected by flow cytometric microsphere-based assay. The colon tissues samples were sectioned， and immunohistochemical method was used to detect the expression levels of T-box transcription factor （T-bet）， GATA binding protein 3 （GATA-3）， IL-17A and forkhead box P3 （FoxP3） in colon tissues. The content of transforming growth factor-β （TGF-β） in colon tissue of mice was determined by enzyme-linked immunosorbent assay （ELISA）. ResultCompared with the normal group， the number of Th1 and Th17 cells in the model group was significantly increased， while the number of Th2 and Treg cells was significantly decreased， and Th1/Th2 and Th17/Treg were significantly increased (P<0.05， P<0.01). The expressions of T-bet， GATA-3， IL-17A and FoxP3 in colon tissues of mice were significantly increased (P<0.05， P<0.01)， and the contents of cytokines IL-2， IL-17A， IL-6， TNF-α and IFN-γ were significantly increased. The contents of IL-4， TGF-β and IL-10 were significantly decreased (P<0.05， P<0.01). Compared with the model group， the imbalance of Th1/Th2 and Th17/Treg cells was reversed in each dose group of Huangqin decoction (P<0.01). The expressions of T-bet and GATA-3 in colon of mice were significantly decreased in low-dose， medium-dose and high-dose groups of Huangqintang (P<0.05， P<0.01)， and the expression of IL-17A was significantly decreased in high-dose and low-dose groups of Huangqintang (P<0.01). The contents of cytokines IL-2， IL-17A， IFN-γ， IL-6 and TNF-α were significantly decreased in the high-dose and medium-dose groups of Huangqintang， and the contents of TGF-β and IL-10 were significantly increased in the high-dose group of Huangqintang (P<0.05， P<0.01). ConclusionHuangqintang could reduce colonic inflammation and improve colonic tissue damage in UC mice， possibly by regulating Th17/Treg and Th1/Th2 balance and related cytokines levels， and the release level of pro-inflammatory cytokines.

OBJECTIVE To study the “property-effect” correlation material basis of the classic famous prescription Taohe chengqi decoction in removing blood stasis and purging heat. METHODS The rat model of blood storage syndrome was established to investigate the effects of Taohe chengqi decoction （17.29 g/kg） and its property grouping components [bitter-cold property group （Rhei Radix et Rhizoma+Natrii Sulfas） 9.43 g/kg， pungent-warm property group （Cinnamomi Ramulus+Persicae Semen） 4.71 g/kg， sweet-flat property group （Glycyrrhizae Radix et Rhizoma） 3.14 g/kg] on coagulation indexes （prothrombin time， prothrombin activity， activated partial thrombin time， fibrinogen， thrombin time and endothelin-1） and inflammation indicators （C- reactive protein， tumor necrosis factor-α and superoxide dismutase） in rats. On the basis of determining the dominant property groups， ultra-performance liquid chromatography-quadrupole-orbitrap high-resolution mass spectrometry was used to identify the components. The molecular docking was performed， and the components with binding energy ≤－5.0 kcal/mol were selected as the basis of “property-effect” correlation material basis. RESULTS Taohe chengqi decoction total prescription group and pungent-warm property group could significantly improve the coagulation indexes of model rats （P＜0.05 or P＜0.01）， and the total prescription group and bitter-cold property group could significantly improve the inflammation indexes of model rats （P＜0.05 or P＜0.01）. The pungent-warm property group was the dominant property group for removing blood stasis，and the bitter-cold property group was the dominant property group for purging heat. Thirty-two and thirty-five components were identified from the pungent-warm property group and bitter-cold property group， respectively. With binding energy ≤－5.0 kcal/mol， there were 10 components （abscisic acid， 3，4-dihydroxypheny- lethanol， procyanidin B1， etc.） in the pungent-warm property group and 13 components （baicalein， aloe-emodin， demethylwedelolactone， etc.） in the bitter-cold property group. CONCLUSIONS Taohe chengqi decoction has the effect of removing blood stasis and purging heat， and its material basis comes from pungent-warm property group （Cinnamomi Ramulus， Persicae Semen） and bitter-cold property group （Rhei Radix et Rhizoma， Natrii Sulfas） respectively.

Objective:To investigate the factors affecting phenotypes in the patients of methylmalonic acidemia combined with homocysteinemia cblC type with MMACHC c. 609G>A homologous variant. Methods:A retrospective study on the clinical manifestations, complications, treatment, and outcome in 164patients of cblC type with MMACHC c. 609G>A homologous variant was conducted.The patients were diagnosed by biochemical and genetic analysisfrom January 1998 to December 2020. Results:Among the 164 patients, 2 cases were prenatally diagnosed and began treatment after birth. They are 3 and 12 years old with normal physical and mental development. Twenty-one cases were diagnosed by newborn screening. Among them, 15 cases had with normal development. They were treated fromthe age of two weeks at the asymptomatic period. Six cases began treatment aged 1 to 3 months after onset. Their development was delayed. One hundred and forty-one cases were clinically diagnosed. Their onset age ranges from a few minutes after birth to 6 years old. 110 cases had early-onset (78.0%). 31 cases had late-onset (22.0%). Five of them died. 24 patients lost to follow-up. Of the 141 clinically diagnosed patients, 130 (92.2%) with psychomotor retardation, 69 (48.9%) with epilepsy, 39 (27.7%) with anemia, 30 (21.3%) had visual impairment, 27 (19.1%) had hydrocephalus, 26 (18.4%) had feeding difficulties, 7 (5.0%) with liver damage, and 5 (3.5%) with metabolic syndrome. The frequency of hydrocephalus and seizures was significantly higher in the early-onset group. The urinary methylmalonic acid increased significantly in the patients with epilepsy. During the long-term follow-up, the level of plasma total homocysteine in the seizure-uncontrolled group was significantly higher than that in the seizure-controlled group, the difference had a statistical significance ( P<0.05). Conclusion:Most of the patients with MMACHC c. 609G>A homozygous variant had early-onset disease, with a high mortality and disability rate. If not treated in time, it will lead to neurological damage, resulting in epilepsy, mental retardation, hydrocephalus, and multiple organ damage. Pre-symptomatic diagnosis and treatment are crucial to prevent irreversible neurological damage. Neonatal screening and prenatal diagnosis are important to improve the outcome of the patients.

Objective:Use the droplet digital PCR (ddRCR) technology to establish, optimize and evaluate the method of EGFR-T790M mutation detection.Methods:The relevant probes and primers were designed for EGFR-T790M mutations. The ddPCR reaction system was established, the optimal annealing temperature was set and the basic performance of the method was tested. On this basis, from January 2019 to October 2019, 72 cell-free DNA (cfDNA) samples from NSCLC patients were collected from Shandong Cancer Hospital Affiliated to Shandong First Medical University, and clinically verified. The consistency of the gene mutation detections with Bole ddPCR products was analyzed using Kappa test.Results:The ddPCR reaction system was established and optimized. Linear evaluation showed the R2 value was greater than 0.99. Using ddPCR, the blank detection limit was determined to be the numbers of mutant droplets≥2, with excellent specificity. For the sensitivity analysis, the lower limit of mutation detection was determined to be at least 0.05%. In the repeatability and inter-assay precision tests, the results had a coefficient of variation( CV)<20%. The relative deviation of the results was within the range of±10% for the accuracy analysis. Using the established T790M mutation detection method, 72 samples from the NSCLC patients were tested for genetic mutation in cfDNA, and the overall agreement with the Bole ddPCR products was 91.67% (66/72, Kappa=0.749; P<0.001). Conclusion:Using ddPCR, the method of EGFR-T790M mutation detection for NSCLC was successfully established.

The Philippines’ medical system is mainly based on the provincial responsibility system and the limited hierarchical. The Philippine government implement Philhealth program which can provide medical insurance for most people. The top 10 fatal diseases in this country includes ischemic heart disease, stroke, lower respiratory tract infection and so on. Of these diseases, the increasing rate of hypertensive heart disease, chronic kidney disease and diabetes are fast. Bone setting, massage and herbal medicine are the major form of traditional medicine in the Philippines. The acceptance of acupuncture and moxibustion by the government and local people is relatively high acupuncture and moxibustion therapy has been included in its medical insurance. There are many limitations on the development of Traditional Chinese Medicine (TCM) theory and Chinese herbal medicine in the Philippines, and the clinical application of acupuncture and Chinese herbal medicine is still limited. TCM education in the Philippines is still not systematic. Therefore, it is suggested to improve the education system of TCM, strengthen the promotion of acupuncture and moxibustion, give full play of the advantages of TCM for native high-risk diseases, and to make use of modern technologies such as telemedicine.

Objective:To analyze the clinical features, genetic characteristics, treatment and follow-up results of patients with hydrocephalus caused by methylmalonic acidemia combined with homocysteinuria, and to discuss the optimal strategies for assessing and treating such patients.Methods:From January 1998 to December 2020, 76 patients with hydrocephalus due to methylmalonic acidemia combined with homocysteinuria in the Department of Pediatrics in 11 hospitals including Peking University First Hospital were diagnosed by biochemical, genetic analysis and brain imaging examination. The patients were divided into operation-group and non-operation-group according to whether they underwent ventriculoperitoneal shunt. The clinical features, laboratory examinations, genotype, and follow-up data were retrospectively analyzed. Data were compared between the two groups using rank sum test, and categorical data were compared using χ 2 test. Results:Among the 76 patients (51 male, 25 female), 5 were detected by newborn screening, while 71 were diagnosed after clinical onset, 68 cases (96%) had early-onset, 3 cases (4%) had late-onset. The most common clinical manifestations of 74 cases with complete data were psychomotor retardation in 74 cases (100%), visual impairment in 74 cases (100%), epilepsy in 44 cases (59%), anemia in 31 cases (42%), hypotonia or hypertonia in 21 cases (28%), feeding difficulties in 19 cases (26%) and disturbance of consciousness in 17 cases (23%). Genetic analysis was performed in 76 cases, all of whom had MMACHC gene variations, including 30 homozygous variations of MMACHC c.609G>A. The most common variations were c.609G>A (94, 62.7%), followed by c.658_660del (18, 12.0%), c.567dupT (9, 6.0%) and c.217C>T (8, 5.3%). Therapy including cobalamin intramuscular injection, L-carnitine and betaine were initiated immediately after diagnosis. A ventriculoperitoneal shunt operation was performed in 41 cases (operation group), and 31 patients improved after metabolic intervention (non-operation group). There was no significant difference in the age of onset, the age of diagnosis, the blood total homocysteine, methionine, and urinary methylmalonic acid concentration between the two groups (all P>0.05). The symptoms of psychomotor development, epilepsy, and visual impairments improved gradually after a long-term follow-up in the operation group. Conclusions:Hydrocephalus is a severe complication of methylmalonic acidemia combined with homocysteinuria. The most common clinical manifestations are psychomotor retardation, visual impairment, and epilepsy. It usually occurs in early-onset patients. Early diagnosis and etiological treatment are very important. Hydrocephalus may improve after metabolic intervention in some patients. For patients with severe ventricular dilatation, prompt surgical intervention can improve the prognosis.

Meridian theory plays an important role in the guidance of clinical practice of acupuncture and moxibustion. Since the publication of(), the meridian theory has been developed. In the paper, in view of complex science, the topological properties of acupoint-symptom network were analyzed quantitatively by taking acupoint as node and indication as the connection, such as high clustering coefficient and the small world effect. It was the first time to give the abstraction for the topological proof of the high efficiency information transmission property of acupoint-symptom network meridian system at different times. Its quantitative and digitalized significance was analyzed on the development of meridian theory under the complex scientific background so as to provide a new thought and method for the study of meridian theory and acupuncture modernization.

Objective: To prepare ibuprofen sustained-release dropping pills, to evaluate the accumulative release percentage in vitro and to study the drug state in the base.Methods: With the drug content, mass ratio of water-soluble base to insoluble base and mass ratio of stearic acid to glyceryl monostearate as the investigation factors, and the comprehensive score of 2-hour and 10-hour cumulative dissolution rate as the evaluation index, a Box-Behnken response-surface method was used to screen the optimal formula of ibuprofen sustained-release dropping pills.The drug state in the matrix was examined by differential scanning calorimetry (DSC).Results: The optimal formula of ibuprofen sustained-release dropping pills was as follows: the drug content of 10%, water-soluble and insoluble matrix ratio of 4∶1, and stearic acid and glyceryl monostearate ratio of 3∶1.The maximum cumulative dissolution rate of ibuprofen sustained-release dropping pills was 78.85%.The DSC analysis showed that the drug crystallization peak disappeared in the sustained-release dropping pills, and formed a solid dispersion.Conclusion: The preparation has good sustained-release effect, and the preparation process is simple.