[The efficacy and safety of nerve loop blocking in the treatment of blepharospasm in Meige syndrome].

Objective: To evaluate the efficacy and safety of nerve loop blocking in the treatment of blepharospasm caused by Meige syndrome. Methods: It was a retrospective case series study. Patients with Meige syndrome characterized by blepharospasm or blepharospasm-oromandibular dystonia who underwent nerve loop blocking in the Ophthalmology Department of Henan No. 3 Provincial People's Hospital from April 2018 to January 2020 were included. Before and after surgery, blepharospasm was graded, and the Burke-Fahn-Marsden Dystonia Rating Scale (BFMDRS) was used to score the symptoms of ocular and oral dystonia. The improvement rate of ocular spasm was calculated after surgery according to the scores. The postoperative complications were observed. The Wilcoxon rank test was used to compare the BFMDRS movement subscale scores before and after surgery. The independent sample Mann-Whitney U test was used to compare the improvement rates of eye spasm between male and female patients and between patients with and without combined oromandibular dystonia. Results: Among the 199 patients included, 64 (32.2%) were males, and 135 (67.8%) were females, aged 58 (51, 64) years (22-79 years). The postoperative follow-up period was 24 (21, 28) months. Preoperatively, blepharospasm was graded as grade 3 in 12 patients (6.0%) and grade 4 in 187 patients (94.0%), while the postoperative blepharospasm grades were grade 0 in 100 patients (50.3%), grade 1 in 64 patients (32.2%), grade 2 in 31 patients (15.6%), and grade 3 in 4 patients (2.0%). There was statistically significant difference in the BFMDRS scores of ocular dystonia before and after surgery [8.0 (8.0, 8.0) vs. 0.0 (0.0, 1.0); Z=-12.41, P<0.001]. The improvement rate of blepharospasm in all patients was 100% (87.5%, 100%), ranging from 43.8% to 100%, with no statistically significant difference between patients of different genders and between patients with and without combined oral dystonia (both P>0.05). Statistically significant difference existed in the scores of oral dystonia before and after surgery [2.0 (0.5, 4.5) vs. 1.0 (0.5, 2.0); Z=-4.38, P<0.001], with 25 of 65 patients (38.5%) having their oral symptoms improved. Postoperative complications included eyelid valgus (7.5%, 15/199), frontal numbness (100%, 199/199) and tearing (89.9%, 179/199). Conclusion: Nerve loop blocking is a relatively safe and effective method in the treatment of blepharospasm symptoms of Meige syndrome.

[Investigation on influenza vaccination policy and vaccination situation during the influenza seasons of 2020-2021 and 2021-2022 in China].

Objective: To investigate the policies and coverage of influenza vaccine during the influenza epidemic seasons of 2020-2021 and 2021-2022 in China. Methods: The national influenza vaccination policy and vaccination rate were investigated in counties and districts and described in the two epidemic seasons. Results: In the epidemic seasons of 2020-2021 and 2021-2022, the vaccination coverage of influenza in China was 3.16% and 2.47%, respectively. The free vaccination policy had the highest vaccination coverage (51.75% and 38.32%), followed by the medical insurance reimbursement policy (9.74% and 7.36%). During the epidemic season of 2021-2022, the number of counties and districts implementing the free vaccination policy in China decreased 61 compared with the previous year, but the number of people covered increased by 51.29%. However, the vaccination coverage of the vast population decreased significantly, with the medical staff (75.69% and 40.15% for two epidemic seasons), preschool children (58.86% and 26.15%), and the elderly (45.71% and 32.94%). During the epidemic season of 2021-2022, the number of counties and districts implementing the medical insurance reimbursement policy increased by 6 compared with the previous year, and the number of people covered increased by 11.12%, but the vaccination coverage decreased. Conclusion: The influenza vaccination rate in China is low, and the implementation of cost preferential policy can greatly improve the influenza vaccination rate.

[Effects of duodenal ligation on gastroesophageal reflux and bleomycin-induced pulmonary fibrosis in rats].

Objective: To investigate the effects of duodenal ligation on gastroesophageal reflux and bleomycin-induced pulmonary fibrosis in rats. Methods: Wistar rats were randomized into the control (Ctrl) group, bleomycin (BLM) group, duodenal ligation (GER) group and duodenal ligation plus bleomycin treatment (BLM+GER) group. At day 0 (d0), duodenum ligation was performed in the GER and the BLM+GER group through an open-abdomen surgery at 1.0 cm below the pylorus by about 30% of the circumference. Meanwhile, sham operation was performed in the Ctrl and the BLM group with similar procedures to the above without ligation of the duodenum. At day 14, bleomycin solution (5 mg/kg, for the BLM and BLM+GER groups) or saline (for the Ctrl and GER groups) was intratracheally instilled. Rats were sacrificed at d28 or at d42. HE, Masson's trichrome or TUNEL staining was performed on lung sections of the groups. The levels of hyrdoxyproline (HYP) or malondialdehyde (MDA) were measured respectively by alkaline hydrolysis or thiobarbituric acid colorimetry. The levels of pepsin and cytokines in bronchoalveolar lavage fluid (BALF) samples were assessed by ELISA. Western blot or RT-PCR was used to quantify relative lung expression of proteins or mRNA, respectively. Results: Lungs of the GER group rats were presented with mild inflammatory cell infiltration. Alveolitis and lung fibrosis was prominent in the BLM group but even more severe in the BLM+GER group. Of the Ctrl, GER, BLM and BLM+GER group, the average numbers of apoptotic cells per each magnified field (×200) on d28 lung sections was (5.6±3.0), (6.4±5.3), (15.4±5.3) and (18.4±9.1), respectively (P=0.008); the proportion (%) of blue-stained area under Masson's trichrome at d42 was (21.5±2.8), (23.4±2.5), (34.0±5.8) and (41.3±2.9) (P<0.05); the HYP contents (mg/L) at d42 of each group was (0.77±0.01), (1.26±0.01), (2.02±0.01) and (2.39±0.01) (P<0.01); the BALF levels of MDA (µmol/L) at d42 were (0.51±0.09), (0.87±0.12), (1.40±0.31) and (1.71±0.12) (P<0.001), and differences of these three indices at d42 reached statistical significance when comparing the Ctrl or GER group with the BLM or BLM+GER group (all P<0.05). The levels of pepsin, pH, interleukin (IL)-1ß, transforming growth factor (TGF)-ß1 and HYP at d28 and d42 were statistically different between the GER group and the Ctrl group (all P<0.05). As compared with the BLM group, the values of TGF-ß1, HYP, p-Smad3, vimentin, p-ERK1/2 and cleaved caspase-3 at d28 and d42 were different in the BLM+GER group (all P<0.05). At both d28 and d42, the BALF levels of pepsin and pH were statistically different between the BLM and the Ctrl group, or between the BLM+GER group and the GER group (all P<0.05). Conclusions: Gastroesophageal reflux is induced through duodenal ligation, which activates proinflammatory and profibrotic signals in the lungs and significantly aggravates bleomycin-induced lung injury and fibrosis. In addition, pulmonary fibrosis may induce or worsen the extent of reflux.

[Correlation between serum bilirubin and cardiovascular autonomic neuropathy in patients with type 2 diabetes].

Objective: To investigate the correlation between serum bilirubin and cardiovascular autonomic neuropathy (CAN) in type 2 diabetes mellitus patients. Methods: A total of 369 patients with type 2 diabetes mellitus who were hospitalized at the Department of Endocrinology, Nanjing Jinling Hospital from April 2017 to October 2018 were enrolled, including 226 males and 143 females, with an average age of (54.6±12.1) years. According to cardiovascular reflex tests (CARTs), all the patients were divided into Non CAN group(149 patients without CAN) and CAN group (220 patients complicated with CAN). The difference of serum bilirubin levels between the two groups was compared. The differences of CARTs and the incidence of CAN were compared by tertiles of serum bilirubin levels. The binary logistic regression was used to analyze the risk factors for diabetic cardiovascular autonomic neuropathy. Results: The serum total bilirubin [(9.28±2.74) µmol/L vs (11.08±2.98) µmol/L, P<0.001], direct bilirubin [(3.17±1.20) µmol/L vs (3.71±1.24) µmol/L, P<0.001] and indirect bilirubin levels [(6.11±1.89) µmol/L vs (7.37±2.10) µmol/L, P<0.001] in CAN group were significantly lower than that in Non CAN group. With the increase of serum bilirubin, the incidence of CAN decreased (P<0.01). Multivariate Logistic regression analysis showed that serum total bilirubin (OR=0.819, 95%CI: 0.744-0.901, P<0.001), direct bilirubin (OR=0.739, 95%CI: 0.601-0.908, P=0.004) and indirect bilirubin (OR=0.749, 95%CI: 0.653-0.860, P<0.001) were inversely correlated with the incidence of CAN. Conclusions: Within the physiological range, lower level of serum bilirubin is inversely correlated with the incidence of CAN. It is noteworthy to screen diabetic cardiovascular autonomic neuropathy in patients with type 2 diabetes mellitus who had a lower serum bilirubin level.

[A cross-sectional study on application of glucocorticoid in systemic lupus erythematosus patients].

Objective: To explore the status of glucocorticoid application in patients with systemic lupus erythematosus (SLE) in China. Methods: Epidemiological survey was used. The SLE patients who met the 1997 classification criteria of American College of Rheumatology were enrolled. The usage of glucocorticoid and related adverse reactions were recorded and analyzed. Results: A total of 400 SLE patients were enrolled, including 35 men and 365 women. The average age was (37.4±14.0) years old, and the average duration of disease was (6.7±5.8) years. There were 310 patients using glucocorticoid as maintenance. Sixty-one percent (n=244) patients started using medium dose (prednisone 30-<60 mg/d) as the initial treatment of glucocorticoid, which lasted for(37±11)days.The time of drug duration in patients with low dose prednisone (7.5-<30 mg/d)and high dose (60-100 mg/d) was(92±20)and(17±3)days respectively (P<0.05 between 3 groups). However, patients receiving different initial dosage were of no discrepancy in the maintenance therapy. During maintenance, even though 51.0% (n=158) patients were on prednisone 2.5-5 mg/d, the duration of drug use in >5-10 mg/d groupwas longer[(29.9±3.3) months]. Patients with involvement of internal organs had a higher tendency to use 60-100 mg/d prednisone or pulse-dose therapy in the initial treatment, nevertheless these two groups had no difference of maintenance dosage. Among all 400 patients, 62 patients withdrew glucocorticoid, including 17 patients with disease remission (4.3%), 44 by self-withdrawal and one with adverse reaction. Conclusion: In China, the medium dosage of glucocorticoid is the most common initial treatment in patients with SLE.Prednisone 2.5-5 mg/d was the most common choicefor maintenance therapy. Currently, the proportion of glucocorticoid withdrawal remains low in SLE patients achieving remission.

[Survey on dietary nutrients intake of Chinese residents between 2010 and 2012].

Objective: To investigate the average dietary nutrients intake status among Chinese residents between 2010-2012, and to compare the difference between rural and urban areas. Methods: Data were collected from Chinese Nutrition and Health Surveillance between 2010-2012. Based on regional features and economic development levels, China's county-level divisions were classified into four types: big city, medium and small city, ordinary rural area and poor rural area. A total of 62 857 valid subjects, including 14 017, 16 539, 19 910 and 12 391 respondents in each group separately, were enrolled by Multi-stage stratified proportion to the population cluster random sampling method. The dietary survey was conducted by recalling the 24 h diet in 3 consecutive days and weighting condiment. The daily dietary intake of energy, macronutrient, dietary fiber, vitamin, and mineral substance per resident were calculated based on China Food Composition. Results: The average energy intake per resident in Mainland China was 9 047.1 kJ, with highest (9 719.3 kJ) in poor rural area and lowest (8 490.4 kJ) in medium and small city. The average dietary protein intake was 64.2 g, with highest (73.9 g) in big city and lowest (60.7 g) in poor rural area. The average dietary fat intake was 79.7 g, with highest (89.4 g) in big city and lowest (70.6 g) in poor rural area. The average intake of micronutrients of vitamin A, vitamin B(2), vitamin C, calcium, iron, phosphorus, zinc was separately 441.9 µg, 0.8 mg, 80.1 mg, 364.3 mg, and 21.4 mg; which were lower than Chinese Dietary Reference Intakes. The percentage of population whose average intake of vitamin A, vitamin B(2), vitamin C, calcium were lower than estimated average requirement was separately 77.0%, 90.2%, 67.7%, and 96.6%. Conclusion: The supply of 3 macronutrient was sufficient in mainland China. However, it was a common issue that there was excessive intake of fat but deficient intake of micronutrients such as vitamin, calcium, iron and zinc. There were differences in nutrition intake among big city, medium and small city, ordinary rural area and poor rural area.

Interaction of Lysinibacillus sphaericus Cry48Aa/Cry49Aa toxin with midgut brush-border membrane fractions from Culex quinquefasciatus larvae.

The Cry48Aa/Cry49Aa mosquitocidal toxin from Lysinibacillus sphaericus was uniquely composed of a three-domain (Cry) toxin and binary (Bin) toxin-like protein, with high toxicity against Culex spp. However, its mode of action against the target mosquitoes is still unknown. In this study, Cry48Aa, Cry49Aa and its N- and C-terminal truncated proteins were expressed and purified, and the binding affinities of the purified proteins with midgut brush-border membrane fractions (BBMFs) from Culex quin-quefasciatus larvae were performed. The results showed that both Cry48Aa and Cry49Aa have specific and high binding affinity to BBMFs, with dissociation constants of 9.5 ± 1.8 and 25.4 ± 3.8 nM, respectively. Competition assays demonstrated that Cry49Aa C-terminal derivatives were able to bind to the BBMFs, whereas Far-Western dot blot analysis revealed that its N-terminal constructs interacted with Cry48Aa. Nevertheless, larvicidal activity was almost lost when Cry49Aa truncated proteins, either individually or in pairs, combined with Cry48Aa. It is concluded that Cry49Aa is responsible for receptor binding and interaction with Cry48Aa and plays an important role in the mechanism of action of these two-component toxins.

Bone-cartilage interface crosstalk in osteoarthritis: potential pathways and future therapeutic strategies.

Currently, osteoarthritis (OA) is considered a disease of the entire joint, which is not simply a process of wear and tear but rather abnormal remodelling and joint failure of an organ. The bone-cartilage interface is therefore a functioning synergistic unit, with a close physical association between subchondral bone and cartilage suggesting the existence of biochemical and molecular crosstalk across the OA interface. The crosstalk at the bone-cartilage interface may be elevated in OA in vivo and in vitro. Increased vascularisation and formation of microcracks associated with abnormal bone remodelling in joints during OA facilitate molecular transport from cartilage to bone and vice versa. Recent reports suggest that several critical signalling pathways and biological factors are key regulators and activate cellular and molecular processes in crosstalk among joint compartments. Therapeutic interventions including angiogenesis inhibitors, agonists/antagonists of molecules and drugs targeting bone remodelling are potential candidates for this interaction. This review summarised the premise for the presence of crosstalk in bone-cartilage interface as well as the current knowledge of the major signalling pathways and molecular interactions that regulate OA progression. A better understanding of crosstalk in bone-cartilage interface may lead to development of more effective strategies for treating OA patients.

Basic science and clinical application of platelet-rich plasma for cartilage defects and osteoarthritis: a review.

Cartilage defects (CDs) and the most common joint disease, osteoarthritis (OA), are characterized by degeneration of the articular cartilage that ultimately leads to joint destruction. Current treatment strategies are inadequate: none results in restoration of fully functional hyaline cartilage, for uncertain long-term prognosis. Tissue engineering of cartilage with auto-cartilage cells or appropriate mesenchymal stem cell (MSC)-derived cartilage cells is currently being investigated to search for new therapies. Platelet-rich plasma (PRP), an autologous source of factors obtained by centrifugation, possesses various functions. For culture of MSCs and cartilage cells, it might be substituted for fetal bovine serum (FBS) with high efficiency and safety. It enhances the regeneration of cartilage cells when added to cartilage tissue engineering constructs for repairing CDs and as regenerative injection therapy for OA. But challenges also remain. Some of the growth factors (GFs) present in PRP have negative effects on the OA joint. It is therefore unlikely that a mix of GFs some of which have negative effects in the OA joint, as present in PRP, will be of benefit in OA. Future directions of PRP application may concentrate on seeking an appropriate and innocuous agent like anti-VEGF antibody that can modulate and control the effect of PRP.

Metabolic changes of human dental pulp after rapid palatal expansion.

OBJECTIVES: To investigate rapid palatal expansion (RPE)-induced metabolic changes in human dental pulp by measuring the expression and activity of aspartate aminotransferase (AST). METHODS: mRNA and protein levels of AST in human dental pulp were measured by quantitative real-time polymerase chain reaction and Western blot, respectively. Furthermore, the activity of AST was measured by a full automatic biochemical analyzer. RESULTS: AST mRNA and protein levels were found to be expressed in normal dental pulp. Moreover, the expression of AST was increased significantly after 14 days of RPE and then decreased at 1 month in retention. Three and 6 months after RPE, the AST expression level was gradually decreased to its baseline level. Similarly, AST activity was significantly elevated after 14 days of RPE, which was then down-regulated at 1 month in retention but was still kept at a higher level as compared with the control group. The enzymatic activity of AST was slowly decreased to its baseline level at 3 and 6 months in retention. CONCLUSIONS: These results showed that significant reversible metabolic changes occurred in dental pulp during RPE, which revealed the high capacity of the pulp tissue for adaptation to this orthopedic method.

Retrospective study on 696 cases of traumatic dental injuries of primary dentition in Xi'an, China.

AIM: The aim of the present study was to explore the epidemiological characteristics and treatment of traumatic dental injuries of primary dentition (TDI-p) in Xi'an, China. MATERIALS: Data of children with TDI-p admitted between January 2018 and December 2019 at the Department of Pediatric Dentistry, Xi'an Jiaotong University Stomatology Hospital were retrospectively analysed. The epidemiological distribution characteristics and related treatment information of TDI-p were described. CONCLUSION: Compared with permanent dentition, TDI-p has its particularities, such as type of trauma, treatment procedure, and behavioural management. More attention should be paid to TDI-p by both dentists and guardians.

MiR-188 inhibits proliferation and promotes apoptosis of lung adenocarcinoma cells by targeting SIX1 to negatively regulate ERK signaling pathway.

OBJECTIVE: To explore the effects of micro ribonucleic acid (miR)-188 on proliferation and apoptosis of lung adenocarcinoma (LUAD) cells, and its potential mechanism. MATERIALS AND METHODS: The expression level of miR-188 in LUAD cell lines was detected via quantitative Real Time-Polymerase Chain Reaction (PCR). The effects of miR-188 overexpression on proliferation and apoptosis of A549 cells were detected using methyl thiazolyl tetrazolium (MTT) assay, colony formation assay, and flow cytometry. The potential targets for miR-188 were predicted using the TargetScan Human database, and the interaction between miR-188 and target gene was determined through Dual-Luciferase reporter assay. Moreover, the associations of miR-188 and sine oculis homeobox homolog 1 (SIX1) with the extracellular signal-regulated kinase (ERK) pathway were detected via Western blotting. RESULTS: The expression of miR-188 significantly declined in LUAD cell lines (p<0.05). The overexpression of miR-188 significantly reduced the proliferation rate of A549 cells and increased the percentage of apoptotic A549 cells (p<0.05). Similarly, it was found in colony formation assay that the overexpression of miR-188 inhibited the colony formation ability of A549 cells most significantly (p<0.05). SIX1 was a direct target for miR-188, and its mRNA and protein expressions were downregulated by the overexpression of miR-188. The remarkable downregulation of phosphorylated ERK was observed in A549 cells overexpressing miR-188, while the decline in phosphorylated ERK was reversed in A549 cells overexpressing miR-188 and SIX1. CONCLUSIONS: The expression of miR-188 is downregulated in LUAD cell lines. The overexpression of miR-188 inhibits proliferation and promotes apoptosis of LUAD cells, whose functional mechanism may be related to its regulation on the ERK signaling pathway by targeting SIX1.

Clinical applications and advances of positron emission tomography with fluorine-18-fluorodeoxyglucose (18F-FDG) in the diagnosis of liver neoplasms.

Positron emission tomography (PET) using 18F-fluorodeoxyglucose (18F-FDG) has been widely used not only for detecting and staging malignant tumours but also for monitoring therapy response and for differentiating malignant lesions from benign lesions. However, the sensitivity of 18F-FDG PET for hepatocellular carcinoma (HCC) is low. Recent research about the novel drug shows that the sensitivity of 11C-acetate PET for HCC is higher than that of 18F-FDG PET and 11C-acetate also indicates tumour cell differentiation. 18F-FDG PET proved to be highly sensitive in detecting hepatic metastases from different primaries. 18F-FDG PET has also been shown to be useful and more accurate than conventional imaging modalities in demonstrating the effect of local treatments in liver tumours. With the development of small animal imaging, animal research about 18F-FDG PET is increasing.

LncRNA H19 polymorphisms associated with the risk of OSCC in Chinese population.

OBJECTIVE: H19, a maternally expressed imprinted gene transcribing a long noncoding RNA, has previously been reported to be involved in tumorigenesis and cancer progression. The relationship between H19 and susceptibility of multiple tumors is inconsistent, and currently there is a lack of association of H19 with the risk of oral squamous cell carcinoma (OSCC). The objective to design this research is to investigate and discuss the association of H19 with susceptibility to oral squamous cell carcinoma in Chinese population. PATIENTS AND METHODS: In this research, Illumina Infinium Human Exome BeadChip technology was used for genetic tests in 362 patients who were pathologically diagnosed with new-onset OSCC and 741 healthy controls with matching gender and age, so as to analyze the association of H19 gene polymorphism sites (rs2735971, rs217727, rs2839698 and rs3024270) with susceptibility of OSCC. RESULTS: The results of this research showed that rs217727 [Additive model: adjusted odds ratio (OR) = 1.22, 95% confidence interval (CI) = 1.02-1.46; AA vs. GG: Adjusted OR = 1.20, 95% CI = 1.00-1.44] was related to the susceptibility of oral squamous cell carcinoma. It was not found in the results that the other three sites were associated with the susceptibility of oral squamous cell carcinoma. CONCLUSIONS: It is indicated in this research that rs217727 is statistically correlated with the susceptibility of OSCC.

Effect of miR-203 expression on myocardial fibrosis.

OBJECTIVE: Cardiovascular disease is one of the diseases threatening human health. Myocardial fibrosis is a major cause of cardiovascular diseases. Studies have shown that over expression of miR-203 can inhibit the fibrosis. Therefore, in this study, the effect of differential expression of miR-203 on fibrosis of cultured mouse cardiomyocytes was investigated. MATERIALS AND METHODS: Activators and inhibitors of miR-203 were designed according to the sequence of miR-203, synthesized, and transfected into mouse cardiomyocytes to establish activator group, inhibitor group, and control group. The expression levels of fibrosis-related factors including FN, CTGF, and TGF-ß1 were measured by Western blot and RT-PCR 24 h and 36 h after transfection. RESULTS: Over-expression of miR-203 in mouse cardiomyocytes significantly decreased the expression levels of TGF-ß1, CTGF, and FN in a time-dependent manner, compared with that in the control group (p <0.05). Inhibition of miR-203 expression in mouse cardiomyocytes significantly increased the expression levels of TGF-ß1, CTGF, and FN 36 h after transfection, compared with that in the control group (p < 0.05). No significant differences were seen in the expression levels of TGF-ß1, CTGF, and FN 24 h after transfection, compared with that in the control group (p >0.05). CONCLUSIONS: Over-expression of miR-203 in mouse cardiomyocytes significantly decreased the expression levels of TGF-ß1, CTGF, and FN, which might be used as a detection index for prediction of fibrosis.

Development and application of Real-time PCR assay for detection of Theilera annulata infection in cattle in Xinjiang Uygur Autonomous Region, China.

Theileria annulata (T. annulata), the causative agent of tropical theileriosis, is a protozoan parasite that also causes lymphoproliferative diseases in cattle. Development of reliable and fast methods are necessary in the epidemiological investigation of T. annulata in ticks and animals. Real-time PCR possesses merits of rapidity, accuracy, reliability, automation and ease of standardization, which is widely used for the detection of blood borne parasites. In this study, species-specific primers and TaqMan probe were designed on the basis of the 18s rRNA gene sequence of T. annulata, and the real-time PCR assay was developed by optimizing the reaction parameter. The performance of real-time PCR was assessed by testing 47 blood samples from cattle and comparing with the results from conventional PCR. The results show that this real-time PCR assay could specifically detect 10 copies DNA of T. annulata, which is 10-fold sensitivity more than conventional PCR. No cross-reactions were observed with Babesia bovis, Babesia bigemina, Trypanosoma evansi and Theileria equi. Of the 47 field samples collected from Xinjiang Uygur Autonomous Region, China, 36.17% were detected by real-time PCR, and 25.53% were found positive for T. annulata infection by conventional PCR. These results indicated that the real-time PCR assay is a useful approach for detecting T. annulata infections and has potential as an alternative tool for ecological and epidemiological surveillance of ovine theileriosis.

Hyaluronan preserves peritoneal membrane transport properties.

BACKGROUND: We have shown that intraperitoneal (i.p.) addition of hyaluronan (HA) in a single dwell study in rat could increase peritoneal fluid removal by decreasing the peritoneal fluid absorption rate. In this study, we investigated the impact of repeated use of HA on peritoneal membrane transport characteristics. METHODS: Twelve male Sprague-Dawley rats received a once-daily i.p. injection of 25 mL 4.25% glucose dialysis solution without (HP group, n = 6) or with 0.025% HA (HA group, n = 6) for 1 week. Forty-eight hours after the last injection, a 4-hour dwell using 25 mL 4.25% glucose dialysis solution with i.p. volume marker and frequent dialysate and blood samplings was performed in each rat as well as in rats that did not receive any injection (control group, n = 8). RESULTS: Although the i.p. volumes were significantly lower in the HP and HA groups compared to the control group, i.p. volume in the HA group was significantly higher than in the HP group. Net ultrafiltration at 4 hours was 5.6 +/- 1.3 mL, 10.2 +/- 1.8 mL, and 13.2 +/- 0.6 mL for the HP, HA, and control group, respectively. The peritoneal fluid absorption rate decreased by 45% in the HA group compared to the HP group. There was no significant difference in peritoneal fluid absorption rate between the HA and the control group. No difference was found in the direct lymphatic absorption rate between the HP and HA groups [0.010 +/- 0.003 mL/minute in the HP group and 0.011 +/- 0.004 mL/min in the HA group] although they were both higher than that of the control group (0.004 +/- 0.001 mL/min). The solute transport rates were in general significantly higher in the HP group compared to the HA and control groups, and there was no significant difference between the latter two groups, except that protein transport rate was significantly lower in the HA group compared to the control group. CONCLUSIONS: The present study suggests that (1) repeated exposure to hypertonic glucose-based dialysis solution results in increased peritoneal solute transport rates, as well as increased peritoneal fluid absorption rates; and (2) these changes, reflecting a highly permeable peritoneal membrane, were ameliorated by repeated i.p. addition of hyaluronan. The similar changes in the direct lymphatic absorption rate in rats that received daily i.p. injection of dialysis solution suggest that direct peritoneal lymphatic absorption was not influenced by hyaluronan.

Phospholipids in dialysate and the peritoneal surface layer.

Dialysate concentration of phospholipids has been used to monitor peritoneal membrane status. However, we recently found that the peritoneum has a surface layer in which phospholipids may be the main constituent. Therefore, in this study, we compared the phospholipids composition of peritoneal dialysate and of the peritoneal surface in rats. Eight male Sprague-Dawley rats were used in the study. Five rats received an intraperitoneal injection of 25 mL 4.25% glucose dialysis solution. After four hours, the rats were killed, and the dialysate was drained completely. Then 20 mL of Folch solution was infused into the peritoneal cavity for 30 seconds and drained completely. The other three rats received the Folch solution without dialysis. The effluent and Folch solution were then processed for phospholipids analysis using high performance thin-layer chromatography (HPTLC). The total phospholipids content was ten times higher in the surface layer than in the dialysate effluent. In the effluent, four clearly different components were seen: lysophosphatidylcholine (LPC), sphingomyelin (SM, 29%), phosphatidylcholine (PC, 66%), and phosphatidylinositol (PI, 4.5%). However, in the surface layer, as well as LPC, SM (20.6%), PHC (47%), and PI (6.3%), two additional components were seen, phosphatidylserine (PS, 17.1%) and phosphatidylethanolamine (PE, 8.9%). The quantity of phospholipids in the peritoneal surface of non dialyzed rats was similar to the total quantity of phospholipids (in effluent and in the peritoneal surface) of dialyzed rats. Our results suggest that: (1) a surface layer is present on the peritoneum; this layer could well be extracted by Folch solution; and, with appropriate incubation time, one can separate the surface layer without damaging the mesothelial cells; (2) the composition of phospholipids in the effluent is different from that in the peritoneal surface layer, which contains membrane phospholipids (PS and PE); (3) shielding from the peritoneal surface may be the main reason for the presence of phospholipids in the dialysate.

Comparison of three chronic dialysis models.

The chronic peritoneal dialysis model is important for understanding the pathophysiology of peritoneal transport and for studying biocompatibility of peritoneal dialysis solutions. In this study, we compared three different chronic peritoneal dialysis models. A peritoneal catheter was placed in 23 male Sprague-Dawley rats, 12 of which had an intact omentum (model 1) and 11 of which received an omentectomy (model 2). Seven other rats, without a catheter, received a daily intraperitoneal injection (model 3). Each rat received a daily infusion of 25 mL of 3.86% glucose dialysis solution either through the catheter (models 1 and 2) or through injection (model 3) for 4 weeks. Then, a 4-hour dwell study using 3.86% glucose solution with an intraperitoneal volume marker and frequent dialysate and blood sampling was performed in each rat. The intraperitoneal volume was significantly lower in all the dialysis groups as compared to a control group (n = 6) in which the rats had no chronic dialysate exposure. The peritoneal fluid absorption rate, as well as the direct lymphatic absorption rate, was significantly higher in the three dialysis groups as compared to the control group. In general, no significant differences were seen in any of the parameters among the three dialysis models. Owing to catheter obstruction, three rats in model 1 and four rats in model 2 were lost during dialysis. Histological examination showed no significant differences among the three dialysis groups. Our results suggest that omentectomy may not be necessary in the chronic peritoneal dialysis model when using dialysate infusion and no drainage. Based on the present study, we think that perhaps model 1 may be the method of choice to test new peritoneal dialysis solutions. However, owing to its simplicity, model 3 could also be used if great care is taken to avoid puncturing the intestine or injecting into the abdominal wall.