[Immunoprotective effect of recombinant peptidyl-prolyl cis-trans isomerase from Babesia microti against B. microti infection in mice].

OBJECTIVE: To evaluate the immunoprotective effect of active immunization with recombinant peptidyl-prolyl cis-trans isomerase from Babesia microti against B. microti infection in mice. METHODS: Female BALB/c mice at 6 weeks of age, each weighing approximately 20 g, were divided into the recombinant protein immunization group, the infection control group and the normal control group, of 25, 18, 15 mice in each group, respectively. Mice in the recombinant protein immunization group were given active immunization with recombinant BmPPIase protein, and 18 mice with the highest antibody titers were intraperitoneally injected with 100 µL of B. microti-infected whole blood 2 weeks after the last immunization. Mice in the infection control group were intraperitoneally injected with 100 µL of B. microti-infected whole blood, while 15 mice in the normal control group received no treatment. Blood samples were collected from mice in the recombinant protein immunization group and the infection control group on days 0 to 30 post-immunization for detection of B. microti infection, and blood samples were collected on days 0, 7, 14, 21, and 28 post-immunization for routine blood tests with a blood cell analyzer and for detection of serum cytokines using cytometric bead array. RESULTS: Anti-BmPPIase antibodies were detected in 25 mice in the recombinant protein immunization group 2 weeks after the last immunization, with titers of 5 × 103 to 8 × 104. B. microti infection rate peaked in mice in both the recombinant protein immunization and the infection control group on day 7 post-immunization, with positive infection rates of 13.3% and 50.0%, and there were significant differences between the two groups in terms of B. microti infection rate on days 3 (χ2= 113.18, P < 0.01), 5 (χ2 = 475.22, P < 0.01), 7 (χ2 = 465.98, P < 0.01) and 9 post-infection (χ2= 18.71, P < 0.01), while the B. microti infection rate tended to be 0 in both groups on day 11 post-immunization. Routine blood tests showed higher red blood cell counts [(5.30 ± 0.50) × 1012 to (9.87 ± 0.24) × 1012 counts/L)] and hemoglobin levels [(89.67 ± 22.80) to (148.60 ± 3.05) g/L)] in the recombinant protein immunization group than in the infection control group on days 0 to 28 post-immunization. Cytometric bead array detected higher serum interferon-γ [(748.59 ± 17.56) to (3 858.28 ± 1 049.10) fg/mL], tumor necrosis factor-α [(6 687.34 ± 1 016.64) to (12 708.13 ± 1 629.79) fg/mL], interleukin (IL)-6 [(611.05 ± 75.60) to (6 852.68 ± 1 554.00) fg/mL] and IL-17a [(167.68 ± 185.00) to (10 849.27 ± 355.40) fg/mL] and lower IL-10 levels [(247.65 ± 138.00) to (18 787.20 ± 2 830.22) fg/mL] in the recombinant protein immunization group than in the infection control group during the study period. CONCLUSIONS: Recombinant BmPPIase protein induces up-regulation of interferon-γ, tumor necrosis factor-α and presents a high immunoprotective activity against B. microti infection in mice, which is a potential vaccine candidate protein.

Advances in the study of molecular identification technology of Echinococcus species.

The larvae of Echinococcus (hydatidcyst) can parasitize humans and animals, causing a serious zoonotic disease-echinococcosis. The life history of Echinococcus is complicated, and as the disease progresses slowly after infection, early diagnosis is difficult to establish. Due to the limitations of imaging and immunological diagnosis in this respect, domestic and foreign scholars have established a variety of molecular detection techniques for the pathogen Echinococcus over recent years, mainly including nested polymerase chain reaction (PCR), multiplex PCR, real-time quantitative PCR, and nucleic acid isothermal amplification technology. In this article, the research progress of molecular detection technology for Echinococcus infection currently was reviewed and the significance of these methods in the detection and diagnosis of hydatid and hydatid diseases was also discussed.

[CircRNA circTNPO1 promotes the proliferation and metastasis of osteosarcoma by sponging miR-338-3p].

Objective: To explore the effects of circTNPO1 on the proliferation and metastasis of osteosarcoma (OS) by sponging miR-338-3p. Methods: The expression of circTNPO1 on osteoblasts and multiple OS cell lines were detected by qRT-PCR. CircTNPO1 stable knockdown 143B cell line was constructed by sh-circTNPO1. Cell count kit 8 (CCK-8) assay and wound healing assay were applied to evaluate the proliferation and metastasis of this cell. Luciferase reporter assay was used to explore the binding between circTNPO1 and miR-338-3p. In xenograft tumor model, miR-338-3p inhibitor or its control was injected into the circTNPO1 knockdown tumors. The weight and size of the tumors were evaluated and Ki-67 expression was detected by immunohistochemistry. Results: The RNA expression of circTNPO1 in OS cell lines U2OS, HOS, MG63, 143B, ZOS and ZOSM were 2.73±0.27, 3.18±0.54, 4.33±0.52, 5.75±0.65, 4.50±0.49 and 3.96±0.35, respectively, higher than 1.00±0.09 in hFOB1.19 (P<0.001). CCK-8 assay revealed that after 48 h and 72 h, the absorbance of sh-circTNPO1 #1 was 0.81±0.05 and 1.09±0.06, while sh-circTNPO1 #2 143B cells was 0.84±0.04 and 1.2±0.04, which were sharply reduced compared with the control (1.00±0.06 and 1.49±0.06, P<0.001); after 48 h and 72 h, the absorbance of 143B cells transfected with circTNPO1 #1 and miR-338-3p (0.92±0.06 and 1.32±0.07) were higher than those of cells transfected with sh-circTNPO1 cells and miR NC (0.92±0.06 and 1.32±0.07, P<0.050). Wound healing assay demonstrated that the 24 hour-migration rates of sh-circTNPO1 #1 and sh-circTNPO1 #2 cells were (24.43±2.15)% and (39.70±4.20)% respectively, which were significantly lower than that of the control [(56.51±3.27)%, P<0.010]; the migration rates of sh-circTNPO1 #1+ miR NC and sh-circTNPO1 #1+ miR-338-3p inhibitor were (26.70±2.21)% and (46.10±5.71)%, with a significant difference (P<0.005). In xenograft tumor model, the weight and size of tumors in control, sh-circTNPO1 #1+ miR NC and sh-circTNPO1 #1+ miR-338-3p inhibitor mice were (458.80±158.10) mg, (262.50±82.09) mg, (395.40±137.60) mg and (593.00±228.40) mm(2,) (203.30±144.20) mm(2,) (488.60±208.60) mm(2,) respectively. Compared with control, sh-circTNPO1 tumors were significantly smaller (P<0.01). Injection with miR-338-3p inhibitor significantly reversed both the weight and size of tumors (P<0.05). Conclusion: CircTNPO1 promotes the proliferation and metastasis of OS by sponging miR-338-3p, which could be a new target for OS treatments.

[Effect of enhanced recovery after surgery on surgical stress response in patients with gastric cancer complicated with type 2 diabetes mellitus].

Objective: To investigate the effect of enhanced recovery after surgery on the stress response of gastric cancer patients complicated with type 2 diabetes mellitus. Methods: We retrospectively analyzed the data of 49 patients with type 2 diabetes who underwent radical gastrectomy for gastric cancer in the Department of gastroenterology of the Affiliated Hospital of Jiangsu University from Jan to Dec 2020. They were randomly divided into experimental group and control group according to different perioperative management measures. The perioperative C-reactive protein (CRP), white blood cell (WBC), interleukin-6(IL-6), insulin resistance (HOMA-IR), blood glucose fluctuation and postoperative recovery were compared between the two groups. Results: A total of 49 patients were enrolled in the study (23 in the experimental group and 26 in the control group). The degree of stress reaction of the experimental group was lighter than that of the control group. The levels of CRP were significantly different on the 5th and 7th day after operation, IL-6 was significantly different on the 1st, 3rd, 5th and 7th day after operation, WBC and HOMA-IR were significantly different on the 1st day postoperatively. And the changes of HOMA-IR and blood glucose in experimental group were more gentle than those in control group. All the differences were statistically significant(P<0.05). In the experimental group, the time of first anal exhaust, indwelling time of drainage tube or nasointestinal tube and the total hospitalization time were significantly shorter than those of the control group(P<0.05). Conclusion: ERAS can reduce the degree of inflammatory stress and the postoperative IR level promote the early recovery of patients with gastric cancer complicated with type 2 diabetes mellitus.

[The expression and clinical significance of long non-coding RNA GIHCG in cholangiocarcinoma].

Objective: To screen long non-coding RNA (lncRNA) related to the prognosis of cholangiocarcinoma patients, detect its expression in cholangiocarcinoma tissue, and analyze its clinical significance by analyzing The Cancer Genome Atlas (TCGA) database. Methods: Using limma package, survival package, and survival receiver operating characteristic curve (ROC) package of R software to analyze the data of cholangiocarcinoma in TCGA and screen the differentially expressed lncRNAs related to patient survival. Real-time PCR and Fish were used to detect the expression of lncRNA and analyze its correlation with the clinical characteristics of patients. Small interfering RNA was used to knock down the expression of lncRNA GIHCG, and its effect on the migration ability of cholangiocarcinoma cell lines was detected by Transwell. Results: The results of the comprehensive analysis of survival, ROC, and correlation analysis with clinical data showed that lncRNA GIHCG has a significant correlation with lymph node metastasis in patients with cholangiocarcinoma. The expression of lncRNA GIHCG in cholangiocarcinoma tissue is significantly increased, closely related to tumor size and lymph node metastasis. Transwell results showed that lncRNA GIHCG could promote the migration of cholangiocarcinoma cells. Conclusion: The expression of lncRNA GIHCG is significantly increased in cholangiocarcinoma tissues and is closely related to patient survival and lymph node metastasis. It is expected to become a new molecular marker for diagnosing or treating cholangiocarcinoma.

[Mechanism of hepatic fibrosis associated with Echinococcus: a review].

Echinococcosis is a zoonotic parasitic disease caused by Echinococcus infections, and this disorder may cause fibrosis of multiple vital organs, which may further progress into cirrhosis. Early-stage hepatic fibrosis is reversible, and unraveling the mechanisms underlying hepatic fibrosis induced by Echinococcus infections is of great significance for the prevention and treatment of early-stage hepatic fibrosis. Recently, the studies pertaining to hepatic fibrosis associated with Echinococcus infections focus on cytokines and immune cells. This review summarizes the advances in the mechanisms underlying host immune cells- and cytokines-mediated hepatic fibrosis in humans or mice following Echinococcus infections.

[Clinical and prognostic study of anti-N-methyl-D-aspartate receptor encephalitis children with paroxysmal sympathetic hyperactivity syndrome].

The data of clinical characteristics, medical cost and prognosis of 22 anti-N-methyl-D-aspartate receptor (anti-NMDAR) encephalitis children from the Department of Neurology, Capital Institute of Pediatrics between May 2018 and January 2021 were analyzed, and 6 of them occurred paroxysmal sympathetic hyperactivity syndrome (PSH). It was found that the anti-NMDAR encephalitis children with PSH had severer consciousness disorder [median Glasgow Coma Scale (GCS) score at admission: 7.5], longer duration of consciousness disorder (median time: 53 days), higher hospitalization cost (median cost: 230 000 RMB), severer neurological injury at onset [median modified Rankin Scale (mRS) score at admission: 4], and longer recovery time of neurological function (median time of mRS score recovered to 0-2: 7 months), compared with those without PSH (all P<0.05). Therefore, more attention should be paid to sympathetic excited symptoms of anti NMDAR encephalitis, and thus identify and intervene early on PSH to reduce the neurological damage and economic burden.

[Construction of a cDNA library for Sparganum mansoni and screening of diagnostic antigen cadidates].

OBJECTIVE: To construct a cDNA library of Sparganum mansoni and immunoscreen antigen candidates for immunodiagnosis of sparganosis mansoni. METHODS: Total RNA was extracted from S. mansoni, and reversely transcribed into cDNA, which was ligated into the phage vector. These recombinant vectors were packaged in vitro to construct the SMART cDNA library of S. mansoni. Then, the cDNA library was immunoscreened with sera from patients with sparganosis mansoni to yield positive clones. The inserted fragments of positive clones were sequenced and subjected to homology analyses, and the structure and functions of the coding proteins were predicted. RESULTS: The SMATR cDNA library of S. mansoni was successfully constructed. The titer of the cDNA library was 6.25 × 106 pfu/mL, with a recombinant efficiency of 100%, and the mean length of the inserted fragments in the library was larger than 1 100 bp. A total of 12 positive clones were obtained by immunoscreening, and were categorized into Sm-I (Sm60-1), Sm-II (Sm58-1), Sm-III (Sm20-1) and Sm-IV (Sm22-3), with 1 134, 1 063, 883 bp and 969 bp long inserted fragments. Their coding proteins were highly homologous with the Spirometra erinaceieuropaei antigenic polypeptide, cytoplasmic antigen, ribosomal protein S4-like protein and unnamed protein product, respectively. CONCLUSIONS: A SMART cDNA library of S. mansoni has been successfully constructed and 4 categories of positive clones have been identified, which provides a basis for further studies on diagnostic antigens for sparganosis mansoni.

Comparison of component separation technique, partition technique, and extended anterolateral thigh flap in complex abdominal wall closure.

PURPOSE: This study tries to compare three methods in complex abdominal wall reconstruction. METHODS: A retrospective review was conducted at a single medical center between December 2008 and May 2019. Forty-seven patients who received abdominal fascia repair were enrolled. The patients were divided into three groups: A [component separation technique (CST)], B (partition technique), and C [extended anterolateral thigh (ALT) flap]. All relevant patient information was collected. Statistical analysis including one-way analysis of variance, Chi-square test, and the receiver operating characteristic curve were used. RESULTS: There were no significant differences between the group results related to gender, age, BMI, follow-up, diabetes mellitus, tobacco, or short-, and long-term complications. However, there were significant differences in fascia defect size between groups (group A: 7.6 cm vs. group B: 10.76 cm vs. group C: 13.64 cm). The averaged operative time in group C (339.25 mins) was significantly longer than that in group A (145.40 mins) and B (152.37 mins). The hospitalization in group C (24.1 days) was significantly longer than that in group A (8.2 days) and B (10.3 days). The complication thresholds of group A and group B are 9.45 cm and 11.75 cm, respectively. CONCLUSION: This study suggests that extended ALT flap provides the largest fascia defect closure, followed orderly by partition technique and CST, but requires longer operative time and hospitalization. There are no significant differences in postoperative complications between three groups. A prospective study with indications based on these findings is suggested.

Hsa_circRNA_101036 acts as tumor-suppressor in oral squamous cell carcinoma cells via inducing endoplasmic reticulum stress.

OBJECTIVE: Endoplasmic reticulum (ER) stress has an effect on cancer cell proliferation and survival. TMTC1 has been reported to be involved in cell proliferation and inflammation, and development of ER. Hsa_circRNA_101036 is an exon circRNA formed by splicing of TMTC1 mRNA precursor. This study intends to explore the effect of hsa_circRNA_101036 on the malignant behavior of oral squamous cell carcinoma through endoplasmic reticulum stress. MATERIALS AND METHODS: We firstly evaluated the levels of Hsa_circRNA_101036 in human oral mucous fibroblasts (hOMF), and in several OSCC cell lines, including FaDu, OECM1, SAS, HSC3. Then, we studied the effects of overexpression of Hsa_circRNA_101036 on the cell proliferation, apoptosis, invasion, migration, and cytokine release in OSCC cells. Finally, we evaluated the levels of CHOP that are critical in ER and the ROS levels in OSCC cells. RESULTS: We found that compared with hOMF, a significantly lower mRNA expression of Hsa_circRNA_101036 was found in OECM1 and HSC3 cells. In OECM1 and HSC3 cells, with overexpression of Hsa_circRNA_101036, a significant decrease in cell proliferation, apoptosis, invasion, migration, and cytokine release was found. A significantly increased ROS, as well as increased protein level of CHOP, P38 and Bcl-2, was found in cells with Hsa_circRNA_101036 overexpression. CONCLUSIONS: This study indicated that Hsa_circRNA_101036 may acts as a tumor suppressor in OSCC via regulating the ER in cancer cells.

Overexpression of XIST facilitates cell proliferation, invasion and suppresses cell apoptosis by reducing radio-sensitivity of glioma cells via miR-329-3p/CREB1 axis.

OBJECTIVE: Glioma is a malignant brain cancer capable of spreading to the microenvironment. Long non-coding RNA (lncRNA) X inactive specific transcript (XIST) was recognized as a significant regulator in many cancers. However, the molecular mechanism of XIST in glioma cell radio-sensitivity requires further exploration. PATIENTS AND METHODS: The expression of XIST, microRNA (miR)-329-3p and cyclic AMP response element-binding protein 1 (CREB1) was evaluated by quantitative Real-time polymerase chain reaction (qRT-PCR). Cell viability and apoptosis were examined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and flow cytometry, respectively. Transwell assay was performed to detect cell invasion. Protein expression of gamma-H2AX (γ-H2AX) and CREB1 was determined by Western blot. The correlation between miR-329-3p and XIST or CREB1 was determined by dual-luciferase reporter assay. Animal models were established by subcutaneously injecting U251 cells transfected with sh-XIST and sh-NC. RESULTS: XIST and CREB1 were overexpressed whereas miR-329-3p was low-expressed in glioma tumors and cells compared with the normal counterparts. XIST knockdown inhibited cell proliferation, invasion and induced cell apoptosis by enhancing cell sensitivity to X-ray radiation in glioma. Then, we discovered that miR-329-3p directly interacted with XIST or CREB1 in glioma. In addition, miR-329-3p inhibitor abolished XIST silencing-induced regulatory effects on cell proliferation, apoptosis, invasion, and radio-sensitivity. Meanwhile, miR-329-3p inhibitor counteracted CREB1 silencing-induced inhibition on cell progression and facilitation on radio-sensitivity in glioma. Moreover, we found that XIST could increase CREB1 expression by sponging miR-329-3p. Animal experiments revealed that XIST silencing restrained tumor growth in vivo. CONCLUSIONS: XIST accelerates cell proliferation, invasion and inhibits cell apoptosis by repressing radio-sensitivity of glioma via enhancing CREB1 expression through sponging miR-329-3p, representing prospective methods for glioma treatment.

[Epidemiological characteristics and risk factors of Blastocystis hominis infection among patients with HIV/AIDS in Fuyang City Anhui Province].

OBJECTIVE: To investigate the prevalence and risk factors of Blastocystis hominis infections among patients with HIV/AIDS in Fuyang City, Anhui Province. METHODS: A cross-sectional study was conducted in Fuyang City, Anhui Province in 2016. The demographic and socioeconomic status, and the lifestyle and production style were collected using a questionnaire survey. B. hominis DNA was detected in subjects'stool samples using a PCR assay, and the CD4+ T lymphocyte count and HIV viral load were measured in the subjects' blood samples. The risk factors of B. hominis infections among patients with HIV/AIDS were identified using univariate and multivariate logistic regression analyses. RESULTS: A total of 398 HIV/AIDS patients were enrolled in this study, with a mean age of 49.3 years, a mean body weight of 55.9 kg and a mean height of 164.4 cm. The prevalence of B. hominis infection was 6.78% in the study subjects, and no gender- (χ2 = 1.589, P = 0.207), education level- (χ2 =0.508, P = 0.776), marital status- (χ2 = 0.419, P = 0.811) or occupation-specific prevalence (χ2 = 2.744, P = 0.615) was detected. Among the patients with HIV/AIDS, there were no significant differences in the age (t = 0.370, P = 0.712), height (t = 1.587, P =0.113), body weight (t = 0.516, P = 0.606), CD4+ T lymphocyte count (t = 1.187, P = 0.230) or HIV viral load (t = 0.193, P =0.496) between B. hominis-infected and uninfected individuals. Dinking non-tap water [OR = 6.554, 95% CI: (1.876 to 22.903)] and keeping dogs [OR = 5.895, 95% CI: (2.017 to 17.225)] were identified as risk factors for B. hominis infection in patients with HIV/AIDS. CONCLUSIONS: The prevalence of B. hominis infection is high in HIV/AIDS patients, and drinking non-tap water and keeping dogs are risk factors for B. hominis infection among HIV/AIDS patients.

[Differentiation of prostatitis and clinically significant prostate cancer in peripheral zone using the prostate imaging-reporting and data system].

Objective: To determine whether clinically significant prostate cancer (PCa) and prostatitis in the peripheral zone can be distinguished using prostate imaging reporting and data system version 1 (PI-RADS V1) and version 2(PI-RADS V2). Methods: Between September 2010 and August 2016, mpMRI data of 77 patients with PCa and 29 prostatitis obtained at 3.0 T were collected in Zhangjiagang Hospital Affiliated to Soochow University. Every lesion was scored according to PI-RADS (V1 and V2), as well as a sum score and a PI-RADS V2 score. The non-parametric Kruskal-Wallis test was used to assess differences between PCa and prostatitis regarding PS3, PS4 and PI-RADS V2 score. The diagnostic performance of PI-RADS V1 and V2 for detection of prostatitis in peripheral zone was compared by analyzing ROC curve. Results: The PI-RADS V1 score for PS3, PS4 and the PI-RADS V2-score were all significantly higher for PCa (PS3:12.1±2.1; PS4:16.2±2.9; V2:4.6±0.8) than for prostatitis (PS3:8.0±0.7; PS4:10.6±1.0; V2:3.0±0.5) (all P<0.01). Of these parameters, PS4 achieved the highest predictive value for the presence of prostatitis with an AUC of 0.937, sensitivity and specificity were 87.0%, 97.0% with a threshold of 12.5. Conclusion: Prostatitis can be differentiated from clinically significant PCa in peripheral zone on mpMRI using PI-RADS system, PS4 achieved better results compared to PS3 and V2.

Second-harmonic imaging microscopy for identifying colorectal intraepithelial neoplasia.

In this study, second-harmonic imaging microscopy was used to monitor precancerous colorectal lesions at different stages. It was found that the morphology of glands and lamina propria in mucosa changes with the progression of colorectal diseases from normal to low-grade intraepithelial neoplasia to high-grade intraepithelial neoplasia and this microscopy has the ability of direct visualization of these warning symptoms. Furthermore, two morphologic variables were quantified to determine the changes of glands and collagen in lamina propria during the development of colorectal intraepithelial neoplasia. These results suggest that second-harmonic imaging microscopy has the potential in label-freely and effectively distinguishing between normal and precancerous colorectal tissues, and will be helpful for early diagnosis and treatment of colorectal diseases.

Multichannel continuous-wave fiber cavity ringdown gas sensing utilizing frequency-shifted interferometry.

We present a multichannel continuous-wave (CW) fiber cavity ringdown (FCRD) gas sensing method based on frequency-shifted interferometry (FSI). This scheme detects gas concentration by measuring the intensity decay rates of continuous light from different ringdown cavities in the spatial domain, unlike conventional FCRD techniques, which measure the decay rates of pulse light in the time domain. This method shares one CW light source, one slow detector, and one slow data collector. In order to illustrate the theory, acetylene gas concentration measurement in a two-channel FSI-FCRD system was experimentally conducted in the range of 0%-1%. A linear relation was established between concentration and absorption loss, which is proportional to the intensity decay rate, and the measurement resolutions of 3.871%/dB and 3.658%/dB were achieved, respectively. The results reveal that the proposed system has the advantages of low cost, high sensitivity, high precision, and good stability in multichannel gas detection.

Label-free detection of fibrillar collagen deposition associated with vascular elements in glioblastoma multiforme by using multiphoton microscopy.

Glioblastoma multiforme (GBM-WHO grade IV) is the most common and the most aggressive form of brain tumors in adults with the median survival of 10-12 months. The diagnostic detection of extracellular matrix (ECM) component in the tumour microenvironment is of prognostic value. In this paper, the fibrillar collagen deposition associated with vascular elements in GBM were investigated in the fresh specimens and unstained histological slices by using multiphoton microscopy (MPM) based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG). Our study revealed the existence of fibrillar collagen deposition in the adventitia of remodelled large blood vessels and in glomeruloid vascular structures in GBM. The degree of fibrillar collagen deposition can be quantitatively evaluated by measuring the adventitial thickness of blood vessels or calculating the ratio of SHG pixel to the whole pixel of glomeruloid vascular structure in MPM images. These results indicated that MPM can not only be employed to perform a retrospective study in unstained histological slices but also has the potential to apply for in vivo brain imaging to understand correlations between malignancy of gliomas and fibrillar collagen deposition.