Engineering of molecularly imprinted cavity within 3D covalent organic frameworks: An innovation for enhanced extraction and removal of microcystins.

The scarcity of selective adsorbents for efficient extraction and removal of microcystins (MCs) from complex samples greatly limits the precise detection and effective control of MCs. Three-dimensional covalent organic frameworks (3D COFs), characterized by their large specific surface areas and highly ordered rigid structure, are promising candidates, but suffer from lack of specific recognition. Herein, we design to engineer molecularly imprinted cavities within 3D COFs via molecularly imprinted technology, creating a novel adsorbent with exceptional selectivity, kinetics and capacity for the efficient extraction and removal of MCs. As proof-of-concept, a new CC bond-containing 3D COF, designated JNU-7, is designed and prepared for copolymerization with methacrylic acid, the pseudo template L-arginine and ethylene dimethacrylate to yield the JNU-7 based molecularly imprinted polymer (JNU-7-MIP). The JNU-7-MIP exhibits a great adsorption capacity (156 mg g-1) for L-arginine. Subsequently, the JNU-7-MIP based solid-phase extraction coupled with high performance liquid chromatography-mass spectrometry achieves low detection limit of 0.008 ng mL-1, wide linear range of 0.025-100 ng mL-1, high enrichment factor of 186, rapid extraction of 10 min, and good recoveries of 92.4%-106.5% for MC-LR. Moreover, the JNU-7-MIP can rapidly remove the MC-LR from 1 mg L-1 to levels (0.26-0.35 µg L-1) lower than the WHO recommended limit for drinking water (1 µg L-1). This work reveals the considerable potential of 3D COF based MIPs as promising adsorbents for the extraction and removal of contaminants in complex real samples.

Comprehensive analysis of the clinical significance, immune infiltration, and biological role of MARCH ligases in HCC.

The membrane-associated RING-CH (MARCH) family, a member of the E3 ubiquitin ligases, has been confirmed by a growing number of studies to be associated with immune function and has been highlighted as a potential immunotherapy target. In our research, hepatocellular carcinoma (HCC) patients were divided into C1 and C2 MARCH ligase-related patterns by the non-negative matrix factorization (NMF) algorithm. Multiple analyses revealed that the MARCH ligase-related cluster was related to prognosis, clinicopathological characteristics, and the tumor immune microenvironment (TIME). Next, the signature (risk score) of the MARCH prognosis was constructed, including eight genes associated with the MARCH ligase (CYP2C9, G6PD, SLC1A5, SPP1, ANXA10, CDC20, PON1, and FTCD). The risk score showed accuracy and stability. We found that the correlations between risk score and TIME, tumor mutation burden (TMB), prognosis, and clinicopathological characteristics were significant. Additionally, the risk score also had important guiding significance for HCC treatment, including chemotherapy, immunotherapy, and transarterial chemoembolization (TACE).

Differential Diagnosis of COVID-19 Pneumonia From Influenza A (H1N1) Pneumonia Using a Model Based on Clinicoradiologic Features.

Objectives: Both coronavirus disease 2019 (COVID-19) pneumonia and influenza A (H1N1) pneumonia are highly contagious diseases. We aimed to characterize initial computed tomography (CT) and clinical features and to develop a model for differentiating COVID-19 pneumonia from H1N1 pneumonia. Methods: In total, we enrolled 291 patients with COVID-19 pneumonia from January 20 to February 13, 2020, and 97 patients with H1N1 pneumonia from May 24, 2009, to January 29, 2010 from two hospitals. Patients were randomly grouped into a primary cohort and a validation cohort using a seven-to-three ratio, and their clinicoradiologic data on admission were compared. The clinicoradiologic features were optimized by the least absolute shrinkage and selection operator (LASSO) logistic regression analysis to generate a model for differential diagnosis. Receiver operating characteristic (ROC) curves were plotted for assessing the performance of the model in the primary and validation cohorts. Results: The COVID-19 pneumonia mainly presented a peripheral distribution pattern (262/291, 90.0%); in contrast, H1N1 pneumonia most commonly presented a peribronchovascular distribution pattern (52/97, 53.6%). In LASSO logistic regression, peripheral distribution patterns, older age, low-grade fever, and slightly elevated aspartate aminotransferase (AST) were associated with COVID-19 pneumonia, whereas, a peribronchovascular distribution pattern, centrilobular nodule or tree-in-bud sign, consolidation, bronchial wall thickening or bronchiectasis, younger age, hyperpyrexia, and a higher level of AST were associated with H1N1 pneumonia. For the primary and validation cohorts, the LASSO model containing above eight clinicoradiologic features yielded an area under curve (AUC) of 0.963 and 0.943, with sensitivity of 89.7 and 86.2%, specificity of 89.7 and 89.7%, and accuracy of 89.7 and 87.1%, respectively. Conclusions: Combination of distribution pattern and category of pulmonary opacity on chest CT with clinical features facilitates the differentiation of COVID-19 pneumonia from H1N1 pneumonia.

[The Optimal Surgical Margins of Nephron-sparing Surgery for Stage T 1b Renal Tumors].

OBJECTIVE: To analyze the pathological characteristics and explore the optimal surgical margins (SM) of nephron-sparing surgery (NSS) for stage T 1b renal carcinoma (4-7 cm) on preoperative imaging. METHODS: The clinical and pathological data of 245 cases of stage T 1b kidney cancer from September 2013 to December 2017 were collected and reviewed retrospectively. The radical nephrectomy (RN) was performed on 174 cases and other 71 cases accepted NSS. There were 158 males and 87 females, with a mean age of 59.6 years and mean tumor size of 5.3 cm. RESULTS: Through postoperative pathological examination, 209 (85.3%) cases were confirmed renal clear cell carcinoma and 219 (89.4%) cases were surrounded with visible peritumoralpseudocapsule (PC). 26 (10.6%) cases of cancerous cells invaded beyond peritumoral PC and into renal parenchyma. The infiltrative depth into renal parenchyma beyond PC was all limited in 3 mm and the cases of ≤1, 1-2 and 2-3 mm were 7 (26.9%), 16 (61.5%) and 3 (11.5%), respectively. Multifocal tumors were discovered in 24 (9.8%) cases. The average resection margin for partial nephrectomy was 5 mm (3-7 mm). CONCLUSION: For stage T 1b renal tumors, NSS is acceptable and a 3 mm of surgical margin is safe and suitable to avoid positive SM.

Intrathoracic infusion therapy with Lentinan and chemical irritants for malignant pleural effusion: a systematic review and meta-analysis of 65 randomized controlled trials.

BACKGROUND: Aderivative of Shiitake mushrooms, Lentinan is used to control malignant pleural effusion (MPE) through intrathoracic infusion. PURPOSE: To determine the clinical response, survival and safety of Lentinan plus chemical irritants, and the optimal combinations with chemical irritants, indication, threshold and optimal regimen for achieving the desired responses. STUDY DESIGN: We performed a new systematic review and meta-analysis following the PRISMA guidelines. METHODS: We collected all randomized controlled trials (RCTs) regarding Lentinan plus chemical irritants from Chinese and English electronic databases (from inception until March 2019). We evaluated their bias risk, synthesized data using meta-analysis, and summarized evidence quality following the Grades of Recommendation Assessment, Development and Evaluation approach. RESULTS: We included 65 RCTs involving 4,080 patients and nine chemical irritants. Most trials had unclear bias risk. Lentinan with cisplatin significantly improved complete response [Risk ratio (RR) = 1.68, 95% confidence intervals (CI) (1.51 to 1.87), p < 0.00001, Fig.3a] and quality of life [RR = 1.51 95% CI (1.41 to 1.62), p < 0.00001, Fig.4], and decreased the risk of treatment failure, myelosuppression, gastrointestinal reaction, and chest pain. For patients with moderate to large volume of the pleural effusion, primary treatment, KPS score ≥ 50-60, or anticipated survival time ≥ 3months, Lentinan (3-4 mg/time, once a week for three to four times) withcisplatin (30-40 mg/m2 or 50-60 mg/m2) significantly improved complete response and decreased failure. Most results were robust and moderate quality. CONCLUSION: The results suggest that Lentinan with chemical irritants, especially cisplatin is beneficial to the patient with MPE, and provide evidence for the indication, threshold, and optimal regimen that may achieve success and decrease failure.

The Hepatorenal Toxicity and Tumor Response of Chemotherapy With or Without Aidi Injection in Advanced Lung Cancer: A Meta-Analysis of 80 Randomized Controlled Trials.

PURPOSE: Chemotherapy-induced hepatorenal toxicity often decreases tolerance for further therapies and results in poor quality of life and prognosis for patients with lung cancer. In this meta-analysis, all related studies were systematically re-evaluated to determine whether Aidi injection relieves hepatorenal toxicity and improves tumor response, and to determine its threshold and the optimal treatment regimen for obtaining the desired responses. METHODS: All studies regarding Aidi injection with chemotherapy were gathered from Chinese and English databases (from inception until January 2019). Their bias risk was evaluated and the data were synthesized using meta-analysis; the quality of evidence of all outcomes was rated by using the Grades of Recommendation Assessment, Development, and Evaluation approach. FINDINGS: Eighty randomized controlled trials containing 6279 patients were included in the study. Most of the trials showed unclear risk of bias. Aidi injection with chemotherapy increased the objective response rate (risk ratio [RR], 1.32; 95% CI, 1.25-1.40) and the disease control rate (RR, 1.15; 95% CI, 1.12-1.17) and resulted in a lower incidence of hepatotoxicity (RR, 0.61; 95% CI, 0.55-0.69) and nephrotoxicity (RR, 0.62; 95% CI, 0.53-0.72) than that of chemotherapy alone. Subgroup analyses showed that treatment with 50 mL per time, 10 to 14 days per cycle, and 2 to 3 cycles of Aidi injection with chemotherapy resulted in a low incidence of hepatorenal toxicity. All of the results were robust, and their quality was moderate. IMPLICATIONS: The moderate evidence indicates that Aidi injection with chemotherapy may improve tumor response and result in a low incidence of hepatorenal toxicity in patients with lung cancer. Aidi injection may relieve hepatorenal toxicity and exhibit an important protective effect against chemotherapy-induced hepatorenal toxicity. Based on the subgroup analysis results, Aidi injection seems to lower the threshold for chemotherapy. Treatment with 50 mL per time, 10 to 14 days per cycle, and 2 to 3 cycles may be the optimal usage for attaining a decrease in hepatorenal toxicity.

Clinical efficacy and safety of aidi injection combination with vinorelbine and cisplatin for advanced non-small-cell lung carcinoma: A systematic review and meta-analysis of 54 randomized controlled trials.

The Aidi injection contains multiple active ingredients, including astragaloside (Re, Rb1, and Rg1), ginsenoside, cantharidin, elentheroside E, and syringin, and it is administered with vinorelbine and cisplatin (NP) to treat non-small-cell lung carcinoma (NSCLC). In this study, we performed a systematic review and meta-analysis to determine the clinical efficacy and safety of the Aidi injection with NP, and the optimal threshold and treatment regimen to produce the desired responses. We collected all studies regarding the Aidi injection with NP for NSCLC from Chinese and English databases (up to April 2019). Risk of methodological bias was evaluated for each study. Data for analysis were extracted using a standard data extraction form. Evidence quality was assessed following the Grading of Recommendations Assessment, Development and Evaluation approach. We included 54 trials containing 4,053 patients for analysis. Combining the Aidi injection with NP significantly increased the objective response rate (odds ratio [OR], 1.32; confidence interval [CI], 1.23, 1.42), disease control rate (OR, 1.14; CI, 1.11, 1.18), and quality of life (OR, 1.80; CI, 1.61, 1.98), with decreased risks of myelosuppression, neutropenia, thrombocytopenia, anemia, gastrointestinal reaction, and liver dysfunction. For patients with a Karnofsky Performance Status score of ≥60, the Aidi injection (50 mL/day, two weeks/cycle, with two to three cycles) treatment with vinorelbine (25 mg/m2) and cisplatin (30-35 mg/m2 or 40-50 mg/m2) might be the optimal regimen for producing the desired tumor response and achieving a good safety level. Most results were robust, and their quality was moderate. The results suggest that administration of the Aidi injection and concomitant NP is beneficial to NSCLC, and provide evidence for the optimal threshold and treatment regimen that may improve tumor response with a good safety level.

Clinical efficacy and safety of synthetic thymic peptides with chemotherapy for non-small cell lung cancer in China: A systematic review and meta-analysis of 27 randomized controlled trials following the PRISMA guidelines.

BACKGROUND: Synthetic thymic peptides (sTPs) are used with chemotherapy to treat non-small cell lung cancer (NSCLC). In this study, we have performed a systematic review and meta-analysis of published trials to confirm the clinical efficacy and safety of sTPs, and determine the optimal types, usages, and sTP/chemotherapy combinations to produce the desired responses. MATERIALS AND METHODS: We collected all studies regarding combined sTP therapy and chemotherapy for NSCLC from the Chinese and English databases (up to October 2018). Bias risk was evaluated for each. Data for meta-analysis was extracted using a pre-designed form. Evidence quality was rated using the Grading of Recommendations Assessment, Development and Evaluation approach. RESULTS: We included 27 randomized controlled trials containing 1925 patients, most with unclear bias risk. Combining sTPs with chemotherapy significantly increased the objective response rate [1.28, (1.13 to 1.45)], disease control rate [1.10, (1.01 to 1.18)], quality of life (QOL) [2.05, (1.62, 2.60)], and 1-year overall survival rate [1.43, (1.15 to 1.78)], with decreased risks of neutropenia, thrombocytopenia, and gastrointestinal reactions. Optimal conditions included treatment in combination with gemcitabine or navelbine and cisplatin, twice a week, with one 3-week cycle. In these conditions, thymosin α1 improved both antitumor immunity and tumor response. Most results had good robustness, and their quality ranged from moderate to very low. CONCLUSIONS: The results suggest that treatment with sTPs, especially thymosin α1, and concomitant chemotherapy is beneficial to the patient, and provide evidence for optimal treatment regimens that may increase patient QOL and survival.

[Comparison of dentinal microcracks formation in severely curved root canal with different Ni-Ti instruments].

PURPOSE: This study was aimed to compare the incidence of dentinal microcracks produced by 3 kinds of Ni-Ti instruments during root canal procedures in severely curved canals. METHODS: Two hundred and forty extracted human molars with mesial roots of 25° to 40° curvatures were selected and divided into A, B, C group, with 80 teeth in each group according to root curvature of 25°-30°ï¼ˆexcluding 25°ï¼‰, 30°-35°,35°-40°. Each of them was prepared with K file to 15#. Then, each group was divided into 4 sub-groups (n=20), one was as control, the others were prepared with WaveOne, ProTaper Next and M3-Pro, respectively. After preparation, all roots were stained with 1% methylene blue for 24 hours. The roots were then sectioned at the most curved plane and 2mm below and above the most curved plane with alow-speed saw under cold water. Stereomicroscope was used to inspect dentinal microcracks and differences between each group were analyzed using Chi-square test with SPSS 20.0 software package. RESULTS: Microcracks were observed in the group of WaveOne, ProTaper Next and M3-Pro. WaveOne system induced more dentinal microcracks compared with ProTaper Next and M3-Pro system (P<0.05), and there was no significant difference between ProTaper Next and M3-Pro system (P>0.05). The number of dentinal microcracks in WaveOne, ProTaper Next and M3-Pro group increased with the increase of root curvature. Except Waveone in group A (25°-30°) and group C (35°-40°), the occurrence of dentinal microcracks in two groups had significant difference (P<0.05), there was no significant difference among other groups (P>0.05). CONCLUSIONS: Compared to WaveOne, ProTaper Next and M3-Pro are more suitable for severely curved canal preparation.

Pioglitazone prescription increases risk of bladder cancer in patients with type 2 diabetes: an updated meta-analysis.

Pioglitazone is widely used for glycemic control in patients with type 2 diabetes mellitus, but evidence regarding the association between pioglitazone and bladder cancer risk is confusing. A systematic search of databases was carried out, and other relevant papers were also identified. Then, the analyses were conducted according to the PRISMA and MOOSE guidelines. After quality assessment, nine datasets from 10 available studies were included on the basis of inclusion criteria. The incidence of bladder cancer among pioglitazone ever users and never users, pooled from four cohort and one randomized studies, were 84.51 and 66.68 per 100,000 person-years, respectively. Nine studies representing 2,596,856 diabetic patients were recognized as eligible for overall study; the result suggested an increased risk of bladder cancer in patients exposed to pioglitazone. A persistent significance was detected after being adjusted by age, gender, and use of other diabetes medications. Subgroup analyses indicated that the significantly increased incidence of bladder cancer was found in men, but not in women. Additionally, the analyses addressing increasing exposure to pioglitazone observed a dose-response relation between exclusive ever use of pioglitazone and bladder cancer in terms of cumulative duration of use and cumulative dosage. With some limitations, our results suggest an increased risk of bladder cancer in diabetic patients using pioglitazone, especially for men with long-term and high-dose exposure. Additional studies are needed to provide more precise evidences to support our results.

[Apoptosis of Kasumi-1 cells induced by puerariae radix flavones and its molecular mechanism].

OBJECTIVE: To explore the effects and the molecular mechanism of puerariae radix flavones (PRF) on acute myeloid leukemia cell line Kasumi-1 cells in vitro. METHODS: Kasumi-1 cells treated by PRF for 48 hours were observed with Wright's and Hoechst 33258 dying. The apoptotic cells were analyzed by flow cytometry with AnnexinV/PI staining. The expression levels of bcl-2, Bim and Caspase-3/-8/-9 protein were assayed by Western blot and the AML1-ETO fusion gene was detected by real-time polymerase chain reaction. RESULTS: PRF could induce Kasumi-1 cells to apoptosis effectively. The proportion of apoptotic cells in 50, 200 and 500 µg/ml PRF treatment groups were (14.1 ± 0.8)%, (17.7 ± 1.3)% and (32.4 ± 1.4)%, respectively, and significantly higher than that of control \[(7.8 ± 0.7)%\]. The relative expression levels of the anti-apoptotic Bcl-2 protein were 0.85 ± 0.05, 0.62 ± 0.07 and 0.43 ± 0.05; the apoptotic Bim protein were 0.21 ± 0.06, 0.39 ± 0.04 and 0.75 ± 0.05; the caspase-3 and caspase-9 were 0.92 ± 0.04, 1.21 ± 0.07, 1.33 ± 0.04 and 0.35 ± 0.05, 0.53 ± 0.03, 0.69 ± 0.07, respectively. Compared to the blank control group, all these changes were significant (P < 0.01). Nevertheless, nearly no changes could be observed on the expression level of AML1-ETO fusion gene and caspase-8 protein. CONCLUSION: Apoptosis of Kasumi-1 cells induced by PRF might correlate to the down-regulation of Bcl-2 protein expression and the activation of caspase-3 and caspase-8 protein in the cells. It seemed that all these effects had no relationship with the AML1-ETO fusion gene.

Protective action of endogenously generated H2S on hypoxia-induced respiratory suppression and its relation to antioxidation and down-regulation of c-fos mRNA in medullary slices of neonatal rats.

We previously reported that exogenous H(2)S played roles in protection of respiratory centers against hypoxic injury in medullary slices of neonatal rats. The protective action of endogenous H(2)S and its relation to antioxidation and down-regulation of c-fos mRNA were investigated in the present study. Perfusion of the slices with l-cysteine (Cys), substrate of cystathionine ß-synthase (CBS, H(2)S synthase), could increase frequency of rhythmic respiratory discharge of the hypoglossal rootlets and prevent respiratory suppression induced by hypoxia, whereas perfusion with hydroxylamine (NH(2)OH, inhibitor of CBS) could postpone recovery of respiration from hypoxic inhibition. NH(2)OH also significantly enhanced hypoxia-induced increase in malondialdehyde (MDA) content of the slices. The hypoxia-induced up-regulation of c-fos mRNA could be markedly antagonized by S-adenosyl-l-methionine (SAM, activator of CBS), but greatly increased by NH(2)OH. Neither NH(2)OH, Cys nor SAM had any effect on expression of bcl-2 mRNA in hypoxic medullary slices. These results indicate that endogenously generated H(2)S was involved in protection of the medullary respiratory centers against hypoxic injury partly via antioxidation and down-regulation of c-fos.

Protective effect of hydrogen sulfide on hypoxic respiratory suppression in medullary slice of neonatal rats.

Hydrogen sulfide (H(2)S) has been shown to play a protective role in injury of cells induced by hypoxia. Little is known however about its effect on medullary hypoxia-induced rhythmic respiratory suppression. In the present study, a decrease in frequency of rhythmic discharge of hypoglossal rootlets was observed in medullary slices of neonatal rats perfused with 95% N(2)-5% CO(2) to produce hypoxia. Perfusion with NaHS (H(2)S donor) prevented the inhibitory effect of hypoxia on the burst activity of the rootlets, whereas such action of NaHS was suppressed by pretreatment with glibenclamide, a blocker of K(ATP) channels. In addition, the increase in malondialdehyde content and the up-regulation of c-fos mRNA expression of the slices induced by hypoxia was significantly reduced by NaHS. These results indicate that exogenous H(2)S may protect the medullary respiratory center against hypoxic injury via activation of K(ATP) channels, reduction of lipid peroxidation and down-regulation of c-fos.

[Inhibitory effect of flavonoids of puerarin on proliferation of different human acute myeloid leukemia cell lines in vitro].

This study was aimed to investigate the inhibitory effect of flavonoids of puerarin (PR) in different concentrations on proliferation of 4 kinds of acute myeloid leukemia (AML) cell lines (Kasumi-1, HL-60, NB4 and U937), and to explore its possible mechanism. The MTT method was used to detected the inhibitory effect of PR on proliferation of AML cell lines. The flow cytometry was adopted to determine the change of cell cycle in vitro. The results showed that a certain concentration of PR could inhibit the proliferation of these 4 cell lines effectively in time-and dose-dependent manners, and the intensity of inhibition on 4 kinds of AML cell lines was from high to low as follows: NB4>Kasumi-1>U937>HL-60. Meanwhile, PR could also change cycle process, cell proportion in G1/G0 phase decreased, cells in S phase increased and Sub-diploid peak also appeared. It is concluded that PR can selectively inhibit the proliferation of 4 AML cell lines and block cell cycle process, especially for NB4 cells.

[Apoptosis of NB4 cells induced by flavonoids of puerarin in vitro].

This study was aimed to investigate the effects of flavonoids of puerarin (PR) on apoptosis of acute promyelocytic leukemia (APL) cell line NB4 cells and its mechanism. The NB4 were treated with PR in vitro, the MTT assay was used to detect the inhibitory effect of PR on cell proliferation. The apoptosis of NB4 cells were detected by flow cytometry labelled with Annexin V/PI. The expressions of pml/rar alpha, bcl-2 and survivin were detected by real time reverse transcription-polymerase chain reaction (real time RT-PCR), the expressions of JNK, p38 MAPK, FasL, caspase 3, caspase 8 were detected by Western blot. The results showed that with the increasing of PR concentrations, the apoptosis rates of NB4 cells were gradually elevated. Simultaneously, the mRNA expression of pml/rar alpha, bcl-2 and survivin decreased, while the protein expression of JNK, FasL, caspase 3 and caspase 8 increased, which presented the positive correlation to PR concentrations. When PR combined with arsenic trioxide (ATO), the expression levels of above mentioned mRNA and protein decreased or increased more significantly. It is concluded that PR can effectively induce the apoptosis of NB4 cells. PR combined with ATO displays synergistic effect. It may be triggered by the activation of JNK signal pathway.

[Expression of acid-sensing ion channels in neurons of trapezoid body and lateral paragigantocellular nuclei in rat brain, and effects of intermittent hypoxia on their expression].

OBJECTIVE: To investigate the expression of three acid-sensing ion channel (ASIC) subtypes termed as ASIC1b, ASIC2a, and ASIC3 in the neurons of trapezoid body and lateral paragigantocellular nucleus of rat brainstem, and the effects of intermittent hypoxia on their expression. METHODS: The intermittent hypoxic rat model was established, of which the blood gas analysis was tested after 12 days. The immunohistochemistry SABC method was performed to test the expression of ASIC1b, ASIC2a, and ASIC3 in neurons of trapezoid body and lateral paragigantocellular nucleus in the control (O2) and intermittent hypoxic (IH) groups of rats. RESULTS: The ASIC1b-, ASIC2a- and ASIC3-positive immunoreactive neurons all could be observed in the nucleus of trapezoid body and lateral paragigantocellular nucleus. The intermittent hypoxia group, the numerical density of ASIC1b-positive immunoreactive neurons (cell/mm3) decreased in the nucleus of trapezoid body (P<0.05), but did not significantly change in the lateral paragigantocellular nucleus (P>0.05), the grey level did not significantly change in both the nucleus of trapezoid body and lateral paragigantocellular nucleus (P>0.05); the numerical density of ASIC2a-positive immunoreactive neurons (cell/mm3) did not significantly change in the nucleus of trapezoid body and lateral paragigantocellular nucleus (P>0.05), the grey level in the nucleus of trapezoid body increased (P<0.05) while it did not significantly change in the lateral paragigantocellular nucleus (P>0.05); the numerical density of ASIC3-positive immunoreactive neurons (cell/mm3) decreased in the lateral paragigantocellular nucleus (P<0.05), but it did not significantly change in the nucleus of trapezoid body (P>0.05), the grey level did not significantly change in the nucleus of trapezoid body and lateral paragigantocellular nucleus (P>0.05). CONCLUSION: The ASIC1b, ASIC2a, and ASIC3 exist in the neurons of trapezoid body and lateral paragigantocellular nucleus in the rat brainstem under normal condition, and their expressions in the two nuclei are different to intermittent hypoxia, which means that the roles of subtype of ASICs in different area may be different in the respiratory effects induced by hypoxia.

[Influence of acid stimulation on expression of acid-sensing ion channel 1a and 3 in type I cells of rat carotid body.].

The experiments were carried out to test whether acid-sensing ion channel 1a and 3 (ASIC1a and ASIC3) were expressed on the primarily cultured type I cells of rat carotid bodies (CBs) and whether the expression of the channels was affected by acid stimulation. The Sprague-Dawley rats of either sex (50-100 g) were used. The CBs were isolated and primarily cultured. The immunofluorescent technique was used to detect the expression of tyrosine hydroxylase (TH), a specific marker of type I cells, in order to identify the type of the cultured cells. The double-label immunofluorescent technique was used to detect the expression of ASIC1a and ASIC3 on the TH-positive type I cells. To detect the influence of acid stimulation on the expressions of ASIC1a and ASIC3, each batch of primarily cultured cells were randomly divided into pH7.3 group (control group), pH6.8 group and pH6.2 group (n=9 in each group). The cells in above three groups were treated with pH7.3, pH6.8 and pH6.2 mediums for 24 h, respectively, and then the mRNA expressions of ASIC1a and ASIC3 in type I cells were detected by semi-quantitative RT-PCR technique. The results showed that more than 93% of the primarily cultured CB cells were TH-positive, indicating that most of the cultured cells were type I cells. Furthermore, all TH-positive cells expressed ASIC1a or ASIC3. After the cells were treated with acid stimulation, the amount of ASIC1a mRNA did not change significantly (P>0.05 vs control group); the amount of ASIC3 mRNA had no significant change in pH6.8 group compared with that in control group, but decreased significantly in pH6.2 group (P<0.01 vs control group, P<0.05 vs pH6.8 group). It is concluded that acid stimulation down-regulates the level of ASIC3 mRNA, but has no effect on the level of ASIC1a mRNA.

[A randomized controlled trial of acupuncture treatment of acute ischemic stroke].

OBJECTIVE: To explore effects of acupuncture on ability of daily living (ALD) and the incidence rate of disability and mortality of the patient of acute ischemic stroke. METHODS: Forty patients with acute ischemic stroke were randomly assigned to an acupuncture group and a control group, 20 cases in each group. The treatment group were treated with acupuncture for 3-4 weeks, 5 times each week, and routine therapy. The control group were treated with routine therapy alone. RESULTS: No statistically significant differences between the two groups in the score of neurological defection, and the incidence rate of disability and mortality at following survey of 3 and 6 months were found. CONCLUSION: Acupuncture is safe and feasible for stroke at early stage.

A new rat model of acute seizures induced by tutin.

Coriaria Lactone (CL) is a mixture that has been used to establish animal models of epilepsy. In this study, we focused on the epileptogenic action of tutin, a pure chemical component derived from CL. Rats were implanted with a guide cannula for microinjection of tutin into either of the lateral cerebral ventricles. Behavioral and electroencephalographic (EEG) changes were investigated for at least 2 h after tutin administration. Injected animals presented behavioral seizures: initially, facial and limbic clonus, and subsequently, tonic-clonic seizures that eventually progressed to status epilepticus. Accompanying the behavioral activities, a variety of EEG patterns were recorded. Spike-and-wave complexes occurred continuously at 3 Hz, with a mean amplitude of approximately 295 microV. Multiple spikes and slow waves occurred repetitively and became more frequent and intense. The amplitude of this EEG pattern was low (approximately 85 microV) at onset and gradually increased to approximately 200 microV. Spikes (8 Hz, approximately 555 microV) and slow waves (3 Hz, approximately 670 microV) occurred periodically at the onset of grand mal seizures. Behavioral and EEG changes induced in rats by tutin demonstrated that this is a potent convulsant, by which a new animal model of status epilepticus was established. This acute seizure model is productive and would be optional for investigation of seizures or status epilepticus.