An Analysis of the Clinical and Radiological Prognostic Factors Affecting the Outcomes of Lumbar Intradiscal Biacuplasty.

Purpose: Intradiscal biacuplasty (IDB) has been proven to be effective for treating lumbar degenerative disc disease (DDD). However, there has not been a reported prognostic factor for IDB. The present study meticulously evaluates the general and radiographic features that may serve as markers for predicting the therapeutic outcome of IDB. Methods: A prospective case series study was conducted, following time-series analysis moving averages models, with forty-one patients suffering from chronic discogenic lower back pain for more than six months. These patients subsequently received lumbar cool radiofrequency IDB and were enrolled in the study. Thirty-seven patients completed follow-up questionnaires at 1, 3, 6, and 12 months. The surgical outcomes were reported using visual analogue scale (VAS), Oswestry disability index (ODI), and the consumption of nonsteroidal anti-inflammatory drugs (NSAID). Furthermore, a univariate analysis was performed to identify prognostic factors associated with pain relief from age, gender, body mass index (BMI), and pre-operative lumbar magnetic resonance imaging reading. Results: Significant reductions were found in estimated VAS and ODI at the post-operative period at 1, 3, 6, and 12 months (P < 0.001). The NSAID dosage was significantly decreased at 3-month and 1-year follow-up (P < 0.05). No procedure-related complications were detected. The prognosis of IDB was not related to disc height, Pfirrmann grading or Modic endplate change. However, disc extrusions were associated with promising outcomes (VAS improvement ≥ 50%) on pain relief (P < 0.05). Conclusion: IDB is a good alternative choice for treating lumbar DDD. Patients with a painful extrusion lumbar disc may gain some benefits after receiving IDB following a period of failed conservative treatment. These findings may also add some references for physicians in the decision making when treating lumbar DDD.

Interaction of palmitate and LPS regulates cytokine expression and apoptosis through sphingolipids in human retinal microvascular endothelial cells.

Studies have implicated saturated fatty acid (SFA) and lipopolysaccharide (LPS) in diabetic retinopathy. Since type 2 diabetes is associated with increases in both SFA and LPS in circulation, we investigated how SFA interacts with LPS to regulate proinflammatory cytokine expression and apoptosis in human retinal microvascular endothelial cells (HRMVECs) and the underlying mechanisms. HRMVECs were challenged with palmitate, a major SFA, LPS or palmitate plus LPS and the expression of proinflammatory cytokines were quantified using real-time PCR and enzyme-linked immunosorbent assay. The interaction between palmitate and LPS on inflammatory signaling and sphingolipid metabolism was demonstrated by immunoblotting and lipidomic analysis, respectively. The effect of palmitate and LPS on apoptosis was also studied by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and histone-associated DNA fragment assays. Results showed that palmitate robustly stimulated the expression of proinflammatory cytokines including interleukin (IL)-6 and IL-1ß, and the combination of palmitate and LPS further upregulated the proinflammatory cytokines by cooperatively stimulating inflammatory signaling pathways. Results also showed that while palmitate stimulated ceramide (CER) production via CER de novo synthesis and sphingomyelin (SM) hydrolysis, addition of LPS further increased CER de novo synthesis, but not SM hydrolysis. The involvement of sphingolipids in the cooperative stimulation by palmitate and LPS on cytokine expression was indicated by the findings that the inhibitor of CER de novo synthesis or SM hydrolysis attenuated the stimulation of IL-6 expression by palmitate and LPS. In addition, our study showed that fatty acid receptors GPR40 and CD36 were involved in the IL-6 upregulation by palmitate and LPS. Furthermore, palmitate induced apoptosis via CER production, but addition of LPS did not further increase apoptosis. Taken together, this study showed that palmitate interacted with LPS to upregulate cytokine expression via free fatty acid receptor-mediated inflammatory signaling and sphingolipid metabolism in HRMVECs. In contrast, the interaction between palmitate and LPS did not further increase apoptosis.

Saturated fatty acid combined with lipopolysaccharide stimulates a strong inflammatory response in hepatocytes in vivo and in vitro.

Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease and consumption of high-fat diet (HFD) is a risk factor for NAFLD. The HFD not only increases intake of saturated fatty acid (SFA) but also induces metabolic endotoxemia, an HFD-associated increase in circulating lipopolysaccharide (LPS). Although it is known that SFA or LPS promote hepatic inflammation, a hallmark of NAFLD, it remains unclear how SFA in combination with LPS stimulates host inflammatory response in hepatocytes. In this study, we performed both in vivo and in vitro experiments to investigate the effect of SFA in combination with LPS on proinflammatory gene expression in hepatocytes. Our animal study showed that feeding low-density lipoprotein-deficient mice HFD enriched with SFA and injection of low-dose LPS cooperatively stimulated IL-6 expression in livers. To understand how SFA and LPS interact to promote IL-6 expression, our in vitro studies showed that palmitic acid (PA), a major SFA, and LPS exerted synergistic effect on the expression of IL-6 in hepatocytes. Furthermore, coculture of hepatocytes with macrophages resulted in a greater IL-6 expression than culture of hepatocytes without macrophages in response to the combination of PA and LPS. Finally, we observed that LPS and PA increased ceramide production by cooperatively stimulating ceramide de novo synthesis, which played an essential role in the synergistic stimulation of proinflammatory gene expression by LPS and PA. Taken together, this study showed that SFA in combination with LPS stimulated a strong inflammatory response in hepatocytes in vivo and in vitro.

Nonsteaming method improves the nutritional value and utilization efficiency of silkworm artificial diets.

Artificial diets for silkworms overcome the seasonal limitations of traditional rearing methods with fresh mulberry leaves. However, the current wet artificial diets, steamed at high temperatures, are not favored by silkworms, and they are cumbersome and challenging to preserve. These conditions adversely affected the development of artificial diet-based sericulture production. In this study, we disinfected dry powder diets with radiation and added distilled water without steaming before use. Then, the nutritional value of finished diets and their impact on silkworm development was assessed. Compared with steamed diets, nonsteamed diets were more attractive to silkworms. Chemical assays showed significantly more essential nutrients for silkworms, including l-ascorbic acid, vitamin B1, vitamin B2, and urease in nonsteamed diets than in steamed diets. Feeding fifth-instar silkworm larvae with nonsteamed diets significantly improved the ammonia utilization efficiency of the diet and increased the cocoon shell rate and diet/silk protein conversion efficiency by 5.9% and 13.3%, respectively. When fed with nonsteamed diets, the abundance of aerobic microorganisms in silkworm intestines increased and the abundance of pathogenic bacteria decreased. Furthermore, the vitality of the silkworm, measured by the dead worm cocoon rate, significantly improved by 16.90%. In summary, preparing sterile wet diets without high-temperature steaming effectively improved the nutritional value of the diet and enhanced silkworm growth.

In-depth understanding the synergisms of Cu atomic clusters on Cu single atoms for highly effective electrocatalytic oxygen reduction reaction and Zn-Air battery.

In this paper, a novel, simple and mild soft template assisted strategy and further carbonization approach has been constructed to the size-tunable preparation of porous Cu-N-C/Surfactant catalysts successfully. Note that the pluronic F127 has a significant influence on the synthesis of porous Cu-N-C/F127 with the atomically dispersed Cu-N4 and adjacent Cu atomic clusters (ACs) than other surfactants owing to their particular non-ionic structure. By combining a series of experimental analysis and density functional theory (DFT) calculations, the synergistic effects between the adjacent Cu ACs and atomically dispersed Cu-N4 are favorable for manipulating the binding energy of O2 adsorption and intermediates desorption at the atomic interface of catalysts, resulting in an excellent electrocatalytic ORR performance with a faster kinetics for Cu-N-C/F127 than those of the Cu-N-C, Cu-N-C/CTAB, Cu-N-C/SDS, and comparable with the commercial Pt/C catalyst. This method not only provides a novel approach for synthesizing highly effective copper based single atom catalysts toward ORR, but also offers an in-depth understanding of the synergisms of adjacent ACs on the Cu single atoms (SAs) for highly effective electrocatalytic ORR and Zn-air Battery.

Mechanism of hyperproteinemia-induced damage to female reproduction in a genetic silkworm model.

Hyperproteinemia is a metabolic disorder characterized by abnormally elevated plasma protein concentrations (PPC) in humans and animals. Here, a genetic silkworm model with high PPC was employed to investigate the effect of elevated PPC on female reproduction. Transcriptomic analysis revealed that high PPC induces downregulation of the ovarian development-related genes and disrupts ovarian sugar metabolism. Biochemical and endocrinal analyses revealed that high PPC increases trehalose and glucose levels in hemolymph and glycogen content in the fat body through activation of the gluconeogenic pathway and inhibition of the Insulin/Insulin-like growth factor signaling pathway-the serine/threonine kinase (IIS-AKT) pathway, thus disrupting characteristic metabolic homeostasis of sugar in the ovary. These resulted in ovarian developmental delay as well as reduced number and poor quality of eggs. Insulin supplementation effectively increased egg numbers by lowering blood sugar. These collective results provide new insights into the mechanisms by which high PPC negatively affects female reproduction and support the potential therapeutic effects of insulin.

Hierarchical porous Fe-N/C@surfactant composites synthesized by a surfactant-assisted strategy as high-performance bifunctional oxygen electrodes for rechargeable zinc-air batteries.

Herein, a soft-template strategy involving the cationic surfactants has been successfully applied to size-controlled synthesis of hierarchical porous Fe-N/C for the first time. Specifically, a small amount of Fe and cationic surfactants can be uniformly doped into the zinc-based zeolite imidazole framework (ZIF-8) crystal particles and the cationic surfactants play a critical role in the formation of hierarchically porous Fe-ZIF-8@surfactant precursors. When the Fe-ZIF-8@surfactant is subsequently pyrolyzed, atomically dispersed Fe-Nx coordination structures can be in-situ converted to Fe-N/C, while the cationic surfactants decompose to form a carbon matrix to encapsulate the active sites, thereby preventing the aggregation of nanoparticles to a certain extent. As a result, the combined Fe nanocrystals and atomically dispersed Fe-Nx in the graphitic carbon matrix generate a synergistic effect to boost the electrocatalytic behaviors with a more positive half-wave potential (0.92 V) for oxygen reduction reaction (ORR) and a lower overpotential (420 mV at 10 mA cm-2) for oxygen evolution reaction (OER). As a proof of concept, the Fe-N/C@TTAB based zinc-air batteries (ZABs) present an outstanding peak power density (107.9 mW cm-2) and a superior specific capacity (706.3 mAh g-1) with robust cycling stability over 900 cycles for 150 h, which are better than the commercial Pt/C + IrO2 based ZABs.

Mechanism of hyperproteinemia-induced blood cell homeostasis imbalance in an animal model.

Hyperproteinemia is a metabolic disorder associated with increased plasma protein concentration (PPC) and is often clinically complicated by malignant diseases or severe infections. At present, however, research on the molecular mechanism underlying high PPC (HPPC) is scant. Here, an animal model of primary hyperproteinemia was constructed in an invertebrate ( Bombyx mori) to investigate the effects of HPPC on circulating blood cells. Results showed that HPPC affected blood cell homeostasis, leading to increased reactive oxygen species levels, and induced programmed cell death dependent on the endoplasmic reticulum-calcium ion signaling pathway. HPPC induced the proliferation of blood cells, mainly granulocytes, by activating the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway. Supplementation with the endocrine hormone active substance 20E significantly reduced the impact of HPPC on blood cell homeostasis. Thus, we identified a novel signaling pathway by which HPPC affects blood cell homeostasis, which differs from hyperglycemia, hyperlipidemia, and hypercholesterolemia. In addition, we showed that down-regulation of gene expression of the hematopoietic factor Gcm could be used as a potential early detection indicator for hyperproteinemia.

LaZnB(5)O(10), the first lanthanum zinc borate.

Lanthanum zinc penta-borate, LaZnB(5)O(10), was synthesized by flux-supported solid-state reaction. It is a member of the LnMB(5)O(10) (Ln = rare earth ion and M = divalent metal ion) structure type. The crystal shows a three-dimensional structure constructed from two-dimensional {[B(5)O(10)](5-)}(n) layers with the lanthanum (coordination number nine) and zinc (coordination number six) ions filling in the inter-layers.

N-(4-Chloro-phen-yl)-2-de-oxy-α-l-ribo-pyran-osylamine.

In the crystal structure of the title compound, C(11)H(14)ClNO(3), inter-molecular hydrogen bonds link mol-ecules in the ab plane, forming layers that stack along the c axis.

Long-term leaching behaviours of cement composites prepared by hazardous wastes.

In order to evaluate the long-term environmental impact of Eco-Ordinary Portland Cement (EOPC) prepared by municipal solid wastes (MSS) and hazardous wastes (HW), consecutive leaching tests with a time span of 180 days were conducted on the EOPC composites in the compact and ground forms under deionized and saline water conditions. The results show that the heavy metals investigated can be classified into three groups according to their leaching behaviours. The concentrations of V, Pb, Ni, Ba, Cd and Zn in the leachate increase with the leaching time, which can be classified into the first group. Cu and Sn are in the second group, and their concentrations increase initially, and decline afterward. Cr and As are in the third group, and their concentrations decline firstly, followed by a clear increase. Besides, a kinetic study was also conducted in the present study, revealing that the leaching behaviours of heavy metals follow a second-order model. Furthermore, our results suggest that the EOPC is resistant to the saline water, but the application of such materials in marine conditions should be paid attention to due to the pollution of arsenic.

LPS and palmitate synergistically stimulate sphingosine kinase 1 and increase sphingosine 1 phosphate in RAW264.7 macrophages.

It has been well established that patients with diabetes or metabolic syndrome (MetS) have increased prevalence and severity of periodontitis, an oral infection initiated by bacteria and characterized by tissue inflammation and destruction. To understand the underlying mechanisms, we have shown that saturated fatty acid (SFA), which is increased in patients with type 2 diabetes or MetS, and LPS, an important pathogenic factor for periodontitis, synergistically stimulate expression of proinflammatory cytokines in macrophages by increasing ceramide production. However, the mechanisms by which increased ceramide enhances proinflammatory cytokine expression have not been well understood. Since sphingosine 1 phosphate (S1P) is a metabolite of ceramide and a bioactive lipid, we tested our hypothesis that stimulation of ceramide production by LPS and SFA facilitates S1P production, which contributes to proinflammatory cytokine expression. Results showed that LPS and palmitate, a major SFA, synergistically increased not only ceramide, but also S1P, and stimulated sphingosine kinase (SK) expression and membrane translocation in RAW264.7 macrophages. Results also showed that SK inhibition attenuated the stimulatory effect of LPS and palmitate on IL-6 secretion. Moreover, results showed that S1P enhanced the stimulatory effect of LPS and palmitate on IL-6 secretion. Finally, results showed that targeting S1P receptors using either S1P receptor antagonists or small interfering RNA attenuated IL-6 upregulation by LPS and palmitate. Taken together, this study demonstrated that LPS and palmitate synergistically stimulated S1P production and S1P in turn contributed to the upregulation of proinflammatory cytokine expression in macrophages by LPS and palmitate.

Poly[mu2-(N-hydroxypyridine-2-carboxamidine)-mu2-nitrato-silver(I)].

In the title complex, [Ag(NO3)(C6H7N3O)]n or [Ag(NO3)(pyaoxH2)] (pyaoxH2 is N-hydroxypyridine-2-carboxamidine), the Ag+ ion is bridged by the pyaoxH2 ligands and nitrate anions, giving rise to a two-dimensional molecular structure. Each pyaoxH2 ligand coordinates to two Ag+ ions using its pyridyl and carboxamidine N atoms, and the OH and the NH2 groups are uncoordinated. Each nitrate anion uses two O atoms to coordinate to two Ag+ ions. The Ag...Ag separation via the pyaoxH2 bridge is 2.869 (1) A, markedly shorter than that of 6.452 (1) A via the nitrate bridge. The two-dimensional structure is fishscale-like, and can be described as pyaoxH2-bridged Ag2 nodes that are further linked by nitrate anions. Hydrogen bonding between the amidine groups and the nitrate O atoms connects adjacent layers into a three-dimensional network.

3,4-Dicyano-phenyl 2,3,4,6-tetra-O-acetyl-α-d-glucopyran-oside.

The title compound, C(22)H(22)N(2)O(10), was prepared by the glycosidation method through nitrite displacement on substituted nitro-phthalonitrile. The mol-ecule contains a benzene ring, two nitrile groups and an acetyl-protected d-glucose fragment which adopts a chair conformation. The absolute configuration was determined by the use of d-glucose as starting material. All substituents of the protected sugar are in equatorial positions, with the exclusive presence of the α-anomer. The crystal packing is stabilized by C-H⋯O and C-H⋯N hydrogen-bonding inter-actions.

[The change and mechanism of apoptosis in the hippocampal CA1 region of rats exposed to intermittent hypoxia].

OBJECTIVE: To investigate the effect of intermittent hypoxia on rat hippocampal oxidative stress and neuron apoptosis in the hippocampal CA1 region. METHODS: Thirty six healthy male Wistar rats were randomly divided into three groups of 12 each: a control (NC) group, an intermittent hypoxia (IH) group and a sustained hypoxia (SH) group. The levels of Malondialdehyde (MDA) and Superoxide dismutase (SOD) were detected by colorimetric method. Western blotting was used to examine the expression of p-JNK and p-c-jun. TUNEL was used to detect the neuron apoptosis in the hippocampal CA1 region. RESULTS: The level of MDA (nmol/mg protein) in the hippocampal CA1 region in IH group (1.61 +/- 0.39) was significantly higher than those in NC group (1.25 +/- 0.29) and in SH group (1.34 +/- 0.24), F = 4.185, P < 0.05; the level of SOD (NU/mg protein) in IH group (45 +/- 13) was significantly lower than those in NC group (58 +/- 12) and in SH group (56 +/- 10), F = 4.338, P < 0.05. There were no significant differences between SH and NC groups in the level of MDA or in the activity of SOD (P all > 0.05). The expression of p-JNK and p-c-jun in IH group were 2.1 and 2.3 times the expression in the NC group respectively. The apoptotic indices of IH group (0.30 +/- 0.16) was significantly elevated as compared with group NC (0.12 +/- 0.07) and SH (0.17 +/- 0.09), F = 7.766, P < 0.01. CONCLUSION: Oxidative stress associated with IH in the hippocampal CA1 region can activate JNK signaling pathway, leading to the apoptosis of hippocampal neuron. This maybe the pathophysiological basis of obstructive sleep apnea hypopnea syndrome with neurobehavioral impairments.

[Effects of Shen-Mai injection on sternohyoid contractile properties in chronic intermittent hypoxia rat].

OBJECTIVE: To investigate the effect of Shen-Mai injection (SMI) on sternohyoid contractile properties in rats with chronic intermittent hypoxia. METHODS: Thirty healthy male SD rats were randomly assigned to three equal groups, the control group (A group), the chronic intermittent hypoxia group (B group) and the SMI group(C group). Rats in B group and C group were exposed to alternating periods of hypoxia and normoxia once per minute for 8 h/d for 5 weeks in order to mimic the intermittent hypoxia of obstructive sleep apnea-hypopnea syndrome (OSAHS) in humans. Isometric contractile properties were determined by electrostimulating the strips of isolated sternohyoid muscles at different frequencies (from 10 Hz to 100 Hz) to observe the changes of the sternohyoid contractile properties. RESULTS: (1) The tension of sternohyoid muscle in A group at different frequencies was (23.2 +/- 5.6), (26.2 +/- 5.0), (35.1 +/- 5.4), (46.0 +/- 8.5), (57.0 +/- 9.9), (69.9 +/- 9.7), (79.2 +/- 9.5), (85.7 +/- 7.6), (87.9 +/- 7.9), and (86.6 +/- 12.4) g/cm(2). The tension of sternohyoid muscle in B group [(19.5 +/- 4.7), (23.8 +/- 4.7), (33.0 +/- 5.1), (45.1 +/- 5.9), (54.2 +/- 7.0), (66.1 +/- 9.1), (74.2 +/- 9.1), (79.7 +/- 9.0), (82.0 +/- 8.4), and (80.7 +/- 11.8) g/cm(2)] was not significantly different from those in A group respectively (all P > 0.05); while the tension of sternohyoid muscle in C group [(30.5 +/- 2.3), (40.0 +/- 5.4), (56.2 +/- 7.6), (72.2 +/- 6.4), (82.0 +/- 5.5), (92.4 +/- 4.6), (98.1 +/- 4.0), (99.2 +/- 7.4), (101.8 +/- 3.9), and (102.2 +/- 4.0) g/cm(2)] was significantly different from those in B group respectively (all P < 0.05). (2) In fatigue test, the tension percentages of sternohyoid muscle in A group at 1, 2, 3, 4, and 5 min were (87.9 +/- 5.7)%, (72.1 +/- 11.5)%, (55.6 +/- 9.6)%, (39.7 +/- 10.7)%, (33.2 +/- 10.2)%. Compared with A group, the tension percentages of sternohyoid muscle in B group at 1, 2, 3, 4, and 5 min [(75.6 +/- 8.5)%, (41.6 +/- 7.3)%, (29.0 +/- 2.7)%, (20.4 +/- 2.9)%, (18.5 +/- 2.5)%, respectively] decreased significantly (all P < 0.05). Compared with B group, the tension percentages of sternohyoid muscle in C group [(87.9 +/- 4.4)%, (67.9 +/- 14.1)%, (48.4 +/- 9.9)%, (38.2 +/- 7.0)%, (33.8 +/- 9.3)%, respectively] increased significantly (all P < 0.05). CONCLUSIONS: Chronic intermittent hypoxia can increase upper airway muscle fatigue. SMI can significantly increase the contractile properties of upper airway muscle and resist the fatigue of upper airway muscle.

[Effect of shenmai injection and aminophylline on small airway smooth muscle cell apoptosis and related gene expression in rats with emphysema].

OBJECTIVE: To investigate the effect of Shenmai Injection (SMI) and aminophylline on small airway smooth muscle cell (SASMC) apoptosis and the Fas/FasL expression in the papain induced emphysema model rats. METHODS: Emphysema model in rat was established by a single intratracheal instillation of papain. Apoptosis and Fas/FasL expression of SASMC were examined by immunohistochemical SABC and TUNEL assay at 1, 3, 5, 7, 15 and 30 days after modelling, and the effect of SMI and aminophylline on them were observed. RESULTS: Fas, FasL expressions in normal SASMC were very low with a positive rate of (2.31 +/- 0.05)% and (1.28 +/- 0.47)% respectively. After papain instillation, the positive rates increased along with the prolonging of instillation time. SMI showed an inhibition on SASMC Fas and FasL expression but aminophylline didn't show. SASMC apoptosis was very low in normal rats with a rate of (0.87 +/- 0.32)%, it also raised after papain instillation and increased progressively along with the instillation time. SMI treatment could lower the apoptosis rate but aminophylline couldn't. CONCLUSION: Fas and FasL participated the SASMC apoptosis modulation in emphysema formation. SMI shows a definite treatment effect on emphysema by influencing the Fas and FasL protein expression and reducing SASMC apoptosis through inhibiting the release of inflammatory mediator.

[Effect of shenmai injection on L-type calcium channel of diaphragmatic muscle cells in rats].

OBJECTIVE: To explore the effect of Shenmai Injection (SMI) on L-type calcium channel of diaphragmatic muscle cells in rats. METHODS: Single diaphragmatic muscle cell of rats was obtained by the acute enzyme isolation method and the standard whole-cell patch clamp technique was used to record the inward peak L-type calcium current (IPLC) and current-voltage relationship curve of diaphragmatic muscle cells of 7 rats, and to compare the effects of SMI in various concentrations on them. RESULTS: When keeping the electric potential at -80 mV, stimulation frequency 0.5 Hz, clamp time 300 ms, stepped voltage 10 mV, and depolarized to +60 mV, 10 microliters/ml of SMI could only cause the mean IPLC of rat's diaphragmatic muscle cells increased from -6.9 +/- 0.6 pA/pF to -7.5 +/- 0.7 pA/pF, the amplification being (9.2 +/- 2.8)%, comparison between those of pre-treatment and post-treatment showed insignificant difference. But when the concentration of SMI increased to 50 microliters/ml and 100 microliters/ml, the mean IPLC increased to -8.4 +/- 0.6 pA/pF and -9.2 +/- 0.6 pA/pF, respectively, and the amplification was (22.4 +/- 1.7)% and (34.6 +/- 4.6)% respectively, showing significant difference to that of pre-treatment (P < 0.05). However, SMI showed no significant effect on maximal activation potential and reversal potential. CONCLUSION: SMI can activate the calcium channel of diaphragmatic muscle cells in rats, increase the influx of Ca2+, so as to strengthen the contraction of diaphragmatic muscle, which may be one of the ionic channel mechanisms of SMI in treating diaphragmatic muscle fatigue in clinical practice.

[Effect of Shen-Mai injection and aminophylline on diaphragmatic muscle cell apoptosis and related gene expression in rats with chronic hypoxia].

OBJECTIVE: To investigate the effect of Shen-Mai injection (SMI) and aminophylline on diaphragmatic muscle cell apoptosis and the Fas/FasL expression in chronic hypoxic rats. METHODS: Seventy-five male Wistar rats were randomly divided into three equal groups, control group (A group), SMI group (B group) and aminophylline group (C group). Then each group was further divided into five subgroups of pre-hypoxia, hypoxia 1 w, 2 w, 3 w and 4 w groups (5 rats each). The concentration of oxygen was (10 +/- 3)%, 7 d/w, 8 h/d for all groups, but only B group and C group received SMI (2 ml/d) and aminophylline (10 mg/kg) respectively. Apoptosis and Fas/FasL expression of diaphragmatic muscle cells were examined by the streptavidin biotin-peroxidase complex (SABC) immunohistochemistry techniques and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay, and Dunnett-t test was employed to compare the effects of SMI and aminophylline. RESULTS: (1) Fas, FasL expression in normal diaphragmatic muscle cells was very low with a positive rate of (2.77 +/- 0.45)% and (2.32 +/- 0.61)%. After hypoxia, the positive rates increased with the time of hypoxia time. SMI showed an inhibition on diaphragmatic muscle cell Fas and FasL expression;after hypoxia 1 w, 2 w, 3 w and 4 w, Fas expression [(6.36 +/- 4.17)%, (9.77 +/- 4.12)%, (18.02 +/- 6.91)% and (21.09 +/- 8.09)%] and FasL expression [(5.32 +/- 6.16)%, (9.58 +/- 3.79)%, (12.01 +/- 8.71)%, (19.43 +/- 10.31)%] in B group were different from those in A group respectively (all P < 0.05). But aminophylline did not show such an effect, the expression of Fas [(10.87 +/- 3.62)%, (24.13 +/- 3.79)%, (35.39 +/- 9.02)%, (39.56 +/- 10.12)%] and FasL [(9.37 +/- 4.07)%, (20.16 +/- 4.88)%, (31.81 +/- 7.07)%, (35.51 +/- 9.13)%] were not significantly different from those in A group respectively (all P > 0.05). (2) Diaphragmatic muscle cell apoptosis was very low in normal rats with a rate of (0.93 +/- 0.29)%, which also increased after hypoxia and the increase was associated with the time of hypoxia. Apoptosis rate was decreased by the administration of SMI, the rates of B group were (5.01 +/- 3.71)%, (9.37 +/- 3.12)%, (14.66 +/- 8.76)%, (18.16 +/- 7.02)%, respectively. Except for the first week, the differences of other weeks were all statistically significant when compared with A groups (all P < 0.05). But the effect of aminophylline was different, as compared to A group, only the apoptosis rate in hypoxia 4 w [(30.92 +/- 11.13)%] of C group being statistically significant different (P < 0.05). CONCLUSIONS: Fas and FasL participated in diaphragmatic muscle cell apoptosis in rats with chronic hypoxia. SMI showed a definite effect on the Fas and FasL protein expression and decreased diaphragmatic muscle cell apoptosis, which contributed to the therapeutic effect on diaphragmatic fatigue caused by hypoxia.