MicroRNA (miRNA) are a class of endogenous single -stranded non-coding RNA, which regulate gene expression post-transcriptionally by combining with the target mRNA and play a vital role in biological and pathological processes including the growth of organism , metabolic regulation, disease prediction and intervention.Thus miRNA detection is of considerable significance in disease diagnosis and the research of miRNA function.Because of the restriction factors about miRNA itself such as short sequence, low abundance and highly homologous , traditional methods for miRNA detection cannot meet the current demands due to the limitations like unsatisfactory sensitivity and complicated operation .This review summarizes the newly development about signal amplification -based methods for miRNA, including the advantages and limitations of all kinds of novel methods , and highlights the future trends as well.

Objective:To explore the factors affecting the antiviral treatment efficacy of acquired immunodeficiency syndrome (AIDS) patients.Methods:A total of 107 patients diagnosed with human immunodeficiency virus (HIV) infection in the clinic of Beihai People′s Hospital from January 2016 to June 2018 were selected.The patients were divided into two groups according to whether they voluntarily accepted traditional Chinese medicine treatment, including treatment group who received highly active anti-retroviral therapy (HAART) and traditional Chinese medicine prescription of Ping Gan Jie Du (42 cases), and control group who were only treated with HAART (65 cases). The virological and immunological responses were compared between the two groups at 48 weeks of treatment. The interleukin-28B (IL-28B) rs12979860 genotypes were measured by using the direct sequencing of polymerase chain reaction products. Logistic regression was used to analyze the influencing factors of antiviral efficacy in AIDS patients.Comparison between groups was performed by independent sample t test、matched sample t test or chi-square test. Results:At week 48 of treatment, 41 (97.62%) of the 42 patients in the HAART plus Ping Gan Jie Du group obtained virological response, while 58 (89.23%) of the 65 patients in the HAART group alone acquired virological response, which was not significantly different ( χ2=0.100, P>0.05). The numbers of CD4 + T lymphocytes increased at week 48 of treatment in the HAART plus Ping Gan Jie Du group and HAART group were (244.32±101.83)/μL and (211.56±112.50)/μL, respectively. The was no statistically significant difference ( t=1.522, P>0.05). Among the 92 patients with IL-28B CC genotype, 88 (95.65%) acquired virological response, while 11 of the 15 patients with non-IL-28B CC genotype acquired virological response, which was not significantly different ( χ2=0.394, P>0.05). And CD4 + T lymphocytes in patients with IL-28B CC genotype increased ((229.72±101.17)/μL), which was higher than that without IL-28B CC genotype ((173.40±89.64)/μL), with statistically significant difference ( t=2.028, P=0.045). Multivariate logistic regression analysis showed that baseline CD4 + T lymphocyte count≤200/μL, IL-28B CC genotype, and treatment plan including protease inhibitor were helpful to improve the antiviral efficacy. Conclusion:Baseline CD4 + T lymphocyte count ≤200/μL, IL-28B CC genotype, and protease inhibitor in HAART regimen are the influencing factors of antiviral efficacy in AIDS patients.

Objective To study the regulation of microRNA - 22(miR - 22)on glycometabolism of hemato-poietic stem cell TF - 1 and its molecular mechanism. Methods TF - 1 cells were cultured for 2 h under hypoxic con-ditions. The expression levels of Glut4 and miR - 22 was detected by quantitative real-time PCR(qRT - PCR). The sgRNA vector of the miR - 22 gene was constructed and miR - 22 gene of TK - 1 cells was knocked out by the CRISPR/ Cas9 technique. Overexpression vectors were constructed,and miR - 22 knocked - out cells were introduced to overexpress miR - 22,the expression of miR - 22 was detected by qRT - PCR and the expression levels of Glut4 and PPAR - γ were detected by qRT - PCR and Western blot. Results Compared with the control group,the expression of miR - 22 in TF - 1 cells decreased (0. 015 ± 0. 000 vs. 0. 056 ± 0. 001)and the expression of Glut4 (0. 351 ± 0. 038 vs. 0. 152 ± 0. 005)and PPAR - γ (0. 421 ± 0. 017 vs. 0. 248 ± 0. 008)increased,when TF - 1 cells were cultured for 2 h under hypoxic conditions,and the differences were statistically significant (all P < 0. 05). Compared with the control group,the expression levels of Glut4 (0. 019 ± 0. 00 vs. 0. 008 ± 0. 000)and PPAR - γ (0. 038 ± 0. 001 vs. 0. 019 ± 0. 000)were significantly increased after miR - 22 gene silencing,and they were significantly decreased (Glut4:0. 005 ± 0. 000 vs. 0. 008 ± 0. 000;PPAR - γ:0. 137 ± 0. 000 vs. 0. 019 ± 0. 000)after overexpression of miR - 22,and the differences were statistically significant (all P < 0. 05). Conclusions It suggests that miR - 22 ex-erts a negative regulation on glycometabolism of hematopoietic stem cell TF - 1 by downregulating the expression of PPAR - γ. A new regulatory factor of TF - 1 glycometabolism and the mechanisms are identified,which has provided new ideas for the targeted medication of diseases induced by hematopoietic stem cell dysfunction.

Gallbladder carcinoma is the most common malignant tumor of the biliary tract system. It is characterized by atypical early clinical manifestations, rapid progress and poor prognosis. There is no specific marker for gallbladder cancer, and its preoperative diagnosis mainly relies on imaging examination, while pet-ct is more advantageous in detecting the metastasis of locally advanced gallbladder cancer. Surgical treatment is still the main treatment, but most patients have lost the opportunity of surgical resection by the time of treatment. In recent years, there have been a lot of researches on the diagnosis and treatment of gallbladder cancer at home and abroad. This paper reviews the diagnosis and treatment of gallbladder carcinoma.

Objective To investigate the relationship between vitamin K2,insulin-like growth factor bind-ing protein 3(IGFBP-3),Omentin-1 and the therapeutic effect on children with idiopathic short stature(ISS),and to build a prediction model.Methods A total of 242 ISS children in Jinan Second Maternal and Child Health Hospital from 2019 to 2021 were selected.All of them received recombinant human growth hormone(rhGH)treatment and were divided into effective group and ineffective group according to the therapeutic effect after 12 months of treatment.The general data,vitamin K2,IGFBP-3 and Omentin-1 in the two groups were analyzed.The influencing factors of ISS children's therapeutic effect were analyzed by Logistic regression model and decision tree model.The predictive performance of two models was analyzed by using receiver oper-ating characteristic(ROC)curve.Results There were statistically significant differences in 25-hydroxy vita-min D[25(OH)D],parathyroid hormone(PTH),thyroid stimulating hormone(TSH),vitamin K2,IGFBP-3,Omentin-1,rhGH dosage and weekly outdoor exercise time between the two groups(P＜0.05).Logistic re-gression showed that PTH(OR=7.011,95％CI:2.456-20.014),vitamin K2(OR=0.605,95％CI:.0.465-0.788),IGFBP-3(OR=0.458,95％CI:0.321-0.654),Omentin-1(OR=0.514,95％CI:0.389-0.679)and rhGH dose(OR=0.563,95％CI:0.445-0.712)]were the influential factors for treatment ineffectiveness in ISS children(P＜0.05).The decision tree model showed that vitamin K2,IGFBP-3 and Omentin-1 were the factors influencing the therapeutic effect of ISS,and IGFBP-3 had the most significant impact.ROC curve re-sults showed that the area under the curve of decision tree model and Logistic regression model were 0.922 and 0.908,respectively,with good classification effect.Conclusion The therapeutic effect of ISS children is in-fluenced by factors such as vitamin K2,IGFBP-3,Omentin-1,and so on,and IGFBP-3 has the most significant impact.Logistic regression model and decision tree model could complement each other so as to provide refer-ence for improving the therapeutic effect of ISS children from different aspects.

Objective: To explore the curative effect and safety of botulintum toxin A (BTX-A) on depressive disorder in patients with Parkinson′s disease (PD). Methods: Forty-two cases of PD with depression prospectively recruited in the Second Hospital Affiliated to Soochow University from August 2016 to November 2018 were divided into two groups: 28 patients in BTX-A group (administered with 100 U BTX-A injection on patients′ eyebrow, forehead, bilateral lateral canthus and temporal region at 20 loci), 14 patients in sertraline (control) group (administered with 50-100 (55.36±14.47) mg/d sertraline). The scores of Hamilton Depression Rating Scale (HAMD), Self-rating Depression Scale (SDS), Hamilton Rating Scale for Anxiety (HAMA), Self-rating Anxiety Scale (SAS) after treatment for 2 weeks, 4 weeks, 8 weeks and 12 weeks were compared with the scores of each emotional rating scale for baseline respectively. Meanwhile, the differences in the scores of each emotional scale between the two treatment groups were compared. In addition, the remission rates of depression and anxiety (defined as HAMD, HAMA scores<7) at each follow-up time point between the two groups were compared to evaluate the efficacy and safety of BTX-A in the treatment of PD patients with depression. Results: The scores of HAMD, HAMA, SDS, SAS in the BTX-A group and the sertraline group reduced compared to baseline after treatment (at the 2nd, 4th, 8th, 12th weeks). The scores of HAMD and SDS in the BTX-A group (HAMD scores: F=12.930, P<0.01; SDS scores: F=5.022, P=0.001) and those in the sertraline group (HAMD scores: F=2.883, P=0.030; SDS scores: F=3.427, P=0.013) were significantly lower compared to baseline, but there was no statistically significant difference in the scores of HAMD and SDS between the two groups (P>0.05). HAMD score showed that the remission rate of depression in the BTX-A group (17.9% (5/28), 35.7% (10/28)) was higher than that of the sertraline group (2/14, 4/14) at the 2nd and 4th weeks. At the 8th and 12th weeks, the remission rate of depression in the sertraline group (7/14, 9/14) was higher than that of the BTX-A group (46.4% (13/28), 53.6% (15/28)). There was no statistically significant difference in remission rate of depression between the two groups at each follow-up time point (P>0.05). There was no statistically significant difference in HAMD scores between males and females in the BTX-A group (P>0.05). Two of the 28 patients in the BTX-A group had frown muscle stiffness, which lasted for two weeks and improved in one month. Two patients in the sertraline group had headache and dizziness, and two patients had dry mouth and nausea, which improved after two weeks. There was no statistically significant difference in the incidence of adverse reactions between the two groups (P=0.197). Conclusion BTX-A intraocular facial muscle injection can significantly improve the depressive symptoms of PD patients, and the effect lasts for a long time, with low incidence of side effects and high safety, which can be considered as a safe and effective new method for PD patients with depressive symptoms.

Objective To explore the curative effect and safety of botulintum toxin A (BTX?A) on depressive disorder in patients with Parkinson′s disease (PD). Methods Forty?two cases of PD with depression prospectively recruited in the Second Hospital Affiliated to Soochow University from August 2016 to November 2018 were divided into two groups: 28 patients in BTX?A group (administered with 100 U BTX?A injection on patients′eyebrow, forehead, bilateral lateral canthus and temporal region at 20 loci), 14 patients in sertraline (control) group (administered with 50-100 (55.36±14.47) mg/d sertraline). The scores of Hamilton Depression Rating Scale (HAMD), Self?rating Depression Scale (SDS), Hamilton Rating Scale for Anxiety (HAMA), Self?rating Anxiety Scale (SAS) after treatment for 2 weeks, 4 weeks, 8 weeks and 12 weeks were compared with the scores of each emotional rating scale for baseline respectively. Meanwhile, the differences in the scores of each emotional scale between the two treatment groups were compared. In addition, the remission rates of depression and anxiety (defined as HAMD, HAMA scores<7) at each follow?up time point between the two groups were compared to evaluate the efficacy and safety of BTX?A in the treatment of PD patients with depression. Results The scores of HAMD, HAMA, SDS, SAS in the BTX?A group and the sertraline group reduced compared to baseline after treatment (at the 2nd, 4th, 8th, 12th weeks). The scores of HAMD and SDS in the BTX?A group (HAMD scores: F=12.930, P<0.01; SDS scores: F=5.022, P=0.001) and those in the sertraline group (HAMD scores: F=2.883, P=0.030; SDS scores:F=3.427, P=0.013) were significantly lower compared to baseline, but there was no statistically significant difference in the scores of HAMD and SDS between the two groups (P>0.05). HAMD score showed that the remission rate of depression in the BTX?A group (17.9% (5/28), 35.7% (10/28)) was higher than that of the sertraline group (2/14, 4/14) at the 2nd and 4th weeks. At the 8th and 12th weeks, the remission rate of depression in the sertraline group (7/14, 9/14) was higher than that of the BTX?A group (46.4% (13/28), 53.6% (15/28)). There was no statistically significant difference in remission rate of depression between the two groups at each follow?up time point (P>0.05). There was no statistically significant difference in HAMD scores between males and females in the BTX?A group (P>0.05). Two of the 28 patients in the BTX?A group had frown muscle stiffness, which lasted for two weeks and improved in one month. Two patients in the sertraline group had headache and dizziness, and two patients had dry mouth and nausea, which improved after two weeks. There was no statistically significant difference in the incidence of adverse reactions between the two groups (P=0.197). Conclusion BTX?A intraocular facial muscle injection can significantly improve the depressive symptoms of PD patients, and the effect lasts for a long time, with low incidence of side effects and high safety, which can be considered as a safe and effective new method for PD patients with depressive symptoms.

Objective:To analyze the dynamic changes of T lymphocytes in patients with COVID-19.Methods:Blood samples were collected from 40 COVID-19 cases and 40 healthy controls in Beihai People′s Hospital from January to February, 2020. The counts of CD4 + T and CD8 + T lymphocytes were detected by flow cytometry. Moreover, the T lymphocyte counts in 24 convalescent patients with two consecutive negative nucleic acid test results were also detected. Results:The leukocytes and lymphocytes in the patients with acute COVID-19 were significantly lower than those in the healthy controls [(4.71±1.54)×10 9 cell/L vs (6.26±1.44)×10 9 cell/L, (1.13±0.41)×10 9 cell/L vs (1.51±0.39)×10 9 cell/L; both P<0.05]. The counts of CD4 + T and CD8 + T lymphocytes in the patients with acute COVID-19 were significantly lower than those in the healthy controls [(447.15±144.42) cell/μl vs (592.83±146.76) cell/μl, (309.35±173.05) cell/μl vs (397.20±136.94) cell/μl; both P<0.05], while no significant difference was observed in the CD4 + /CD8 + T cell ratio ( P>0.05). In the 24 convalescent COVID-19 patients, the counts of CD4 + T and CD8 + T lymphocytes were higher during convalescence than in the acute phase [(598.08±138.71) cell/μl vs (420.67±147.38) cell/μl, (439.08±166.94) cell/μl vs (296.67±151.06) cell/μl; both P<0.05], but there was no significant difference in the T lymphocyte counts between the convalescent patients and the healthy controls ( P>0.05). Conclusions:A transient immune deficiency occurred in patients with acute COVID-19, but the impaired immune function could restore to normal level during recovery.

Objective: To explore the improvement rate of liver fibrosis in patients with chronic hepatitis B virus infection who received entecavir alone or in combination with anluohuaxianwan for 78 weeks. Methods: Patients with chronic HBV infection were randomly treated with entecavir alone or in combination with anluohuaxian for 78 weeks. Ishak fibrosis score was used for blind interpretation of liver biopsy specimens. The improvement in liver fibrosis condition before and after the treatment was compared. Student's t test and non-parametric test (Mann-Whitney U-Test and Kruskal-Wallis test) were used to analyze the measurement data. The categorical variables were analyzed by Chi-square test method and Spearman’s rank correlation coefficient was used to test bivariate associations. Results: Liver fibrosis improvement rate after 78 weeks of treatment was 36.53% (80/219) and the progression rate was 23.29% (51/219). The improvement of liver fibrosis was associated to the degree of baseline fibrosis and treatment methods (P < 0.05). The improvement rate of hepatic fibrosis in patients treated with anluohuaxianwan combined with entecavir at baseline F < 3 (54.74%, 52/95) was significantly higher than that in patients treated only with entecavir (33.33%, 16/48), P = 0.016 and the progression rate of hepatic fibrosis (13.68%, 13/95) was lower than that in patients treated alone (18.75%, 9/48), P = 0.466. In patients with baseline F < 3, the proportion of patients with improved and stable liver fibrosis in the combined treatment group (68.1%, 32/47) was higher than that in the treatment group alone (51.7%, 15/29). Conclusion Combined anluohuaxianwan and entecavir treatment can significantly improve the improvement rate of liver fibrosis in patients with chronic hepatitis B virus infection. Furthermore, it has the tendency to improve the stability rate and reduce the rate of progression of liver fibrosis.