Construction and evaluation of a universal influenza mRNA vaccine / 陆军军医大学学报

Objective To construct a universal influenza mRNA vaccine and evaluate its immunogenicity.Methods The antigen sequence of hemagglutinin(HA),nucleoprotein(NP)and matrix protein 2 ectodomain(M2e)in influenza A/California/04/2009 was optimized.HA,NP and 3 tandem M2e(3M2e)were cloned into pcDNA3.1 vector,respectively.Then the mRNAs were synthesized by linearization,in vitro transcription,enzymatic capping and enzymatic tailing,and named as mRNA-HA,mRNA-NP and mRNA-3M2e,respectively.The protein expression of the 3 kinds of mRNAs in 293T cells was detected by immunofluorescence assay.Comb-mRNA vaccine was prepared by enveloped mRNA-HA,mRNA-NP and mRNA-3M2e with lipid nanoparticles,respectively,and the particle size and potential were identified.Twenty-eight 6-week-old female BALB/c mice(18～22 g)were randomly divided into LNP group(n=14)and Comb-mRNA group(n=14).Hemagglutination inhibition(HI)method and microneutralization(MN)test were used to evaluate the serum antibody titer induced by Comb-mRNA vaccines.The mice were infected by 5LD50 wild-type H1 N1 influenza virus to evaluate the protective efficacy.Results The mRNA-HA,mRNA-NP and mRNA-3M2e were successfully constructed,and the 3 mRNAs could be expressed in 293T cells.The average size of mRNA encapsulated by lipid nanoparticles was 119.53±6.5 nm,and the average potential was-8.23±1.3 mV.The geometric mean titer(GMT)of HI and MN in the Comb-mRNA group were 179.6 and 201.6,compared with the LNP group.The ratio of IFN-γ+CD4+/CD8+Tcells was increased.The Comb-mRNA group could provide protection against 5LD50 wild type influenza H1 N1 virus after 2 weeks of booster immunization.Conclusion Comb-mRNA,an influenza vaccine candidate,can induce immune responses and protect mice from influenza virus challenge.

Bilateral hegu acupoints have the same effect on the heart rate variability of the healthy subjects.

Background. The specificity of acupuncture points (acupoints) is one of the key concepts in traditional acupuncture theory, but the question of whether there is adequate scientific evidence to prove or disprove specificity has been vigorously debated in recent years. Acupoint laterality is an important aspect of acupoint specificity. Data is particularly scarce regarding the laterality of the same channel, namesake acupoint located on opposite sides of the body. Our previous study results suggest that Neiguan acupoint (PC6) has the laterality. The aim of this study was to investigate whether Hegu (LI4) also has laterality from the perspective of heart rate variability. Methods. A total of twenty-eight healthy female volunteers were recruited for this study and were randomly separated into the group I (n = 14) and the group II (n = 14) according to the register order. In the group I, left LI4 was stimulated in the first epoch and the right LI4 was stimulated in the second epoch. In the group II, right LI4 was stimulated in the first epoch and left LI4 was stimulated in the second epoch. Electrocardiogram was recorded and heart rate variability was analyzed. Results. The results show that there were no significant differences of heart rate variablity between the group I and the group II in the time domain and in the frequency domain. Conclusions. Bilateral Hegu acupoints have the same effect on the heart rate variability of the healthy subjects.

Change of blood perfusion in Hegu acupoint after contralateral Hegu acupoint was stimulated.

OBJECTIVES: The objective of this study was to investigate the change of mean blood flux in Hegu acupoint after contralateral Hegu acupoint was stimulated. METHODS: Healthy volunteers (N=140) were placed in a temperature-controlled room as a resting state for 60 minutes. Then the measurements of skin blood flow were carried out in pre- and postacupuncture stimulation every 30 minutes over a total of 180 minutes using a Moor full-field laser perfusion imager. The change ratio of mean blood flux in Hegu acupoint was used to evaluate the contralateral stimulation effect in different groups. RESULTS: After acupuncture of the right Hegu acupoint (AR), the change ratio of mean blood flux in the left Hegu acupoint was increased significantly compared with the control group 60 minutes after acupuncture stimulation (p<0.01). Then the blood flow in the AR group was sustained at a higher level compared with that of the control group (p<0.05). However, the change ratio of mean blood flux in the right Hegu acupoint increased significantly in the 30 minutes after contralateral acupuncture stimulation (p<0.01). From that point on, the change ratio was sustained at a higher level in the AR group than in the control group until 150 minutes after stimulation (p<0.05). CONCLUSIONS: On stimulation of either the left or right side of Hegu acupoint, the change ratio of mean blood flux was increased in the contralateral Hegu acupoint in a period from 30, 60, to 150 minutes after stimulation, respectively.

Color spectrum of cupping mark detected by hyperspectral camera:a preliminary observation / 中国针灸

<p><b>OBJECTIVE</b>To perform quantitative observation on the color change of local skin after cupping, so as to explore objective and quantitative methods for skin response of cupping.</p><p><b>METHODS</b>Seven health subjects were included. By quantitative meridian cupping instrument, cupping methods with four types of pressures were respectively performed on subjects for 5 min.The spectrum of cupping mark before and after the cupping was collected by hyperspectral camera, and the color change was recorded by digital camera.</p><p><b>RESULTS</b>Before the cupping, the differences of back skin areas were not significant (>0.05), and its average spectrum indicated two peaks at 540-550 nm and 580-590 nm. After cupping with different pressures, spectrum changes of skin were observed. For -0.02 MPa, the most significant reduction was observed at 550 nm (-12.1%,<0.05); for -0.03 MPa, the most significant reduction was observed at 540 nm (-22.1%,<0.05); for -0.04 MPa, the most significant reduction was observed at 610 nm (-26.7%,<0.05); for -0.05 MPa, the most significant reduction was observed at several spectrums (all<0.05).</p><p><b>CONCLUSIONS</b>After cupping with different negative pressures, significant changes of spectrum are observed on skin; for different pressures, the spectrums of the most significant changes are different; the hyperspectral camera could be applied to perform quantitative observation on the color change of local skin.</p>

Preliminary observation of the blocking effect produced by injecting polyacrylamide hydrogel on low hydraulic resistance channel along meridian / 生物医学工程学杂志

To study the role of meridians in the formation and development of diseases, a pathological model of obstructed channel was established by injecting polyacrylamide hydrogel. The effects of blocking low hydraulic resistance channel (LHRC), produced by injecting polyacrylamide hydrogel and by injecting normal saline, were compared by examining the change in transmission of interstitial fluid pressure wave. The results showed that there was significant decrease (P < 0.01) in interstitial fluid pressure wave after more than 0.5 ml polyacrylamide hydrogel was injected into the channel, whereas no significant changes were found after normal saline was injected or when the hydrogel was outside the channel. The above findings demonstrate that the low hydraulic resistance channel can be blocked by injecting certain amount of polyacrylamide hydrogel and a pathological model of obstructed channel has been established preliminarily.

Influence of Mechanical Pressure to Change of Transcutaneous CO2 Emission by Acupuncture / 针灸推拿医学（英文版）

Objective: In the authors' previous study, acupuncture had an effect along meridians. The effect ofacupuncture can be represented by local energy metabolism which can be expressed by transcutaneous CO2 emission (TCE). To study the transportation of the effect of acupuncture and the influence to the transportation when blocking the meridians, the following experiment was carried out. Methods: Twenty healthy volunteers (male 8, female 12, average age 29) were examined on pericardium meridian at points between Quze (PC 3) and Tianquan (PC 2) (Point a), between Ximen (PC 4) and Quze (PC 3) (Point b), two control points (c and d) beside a and b and Quze (PC 3). TCE was measured on these five points before, during and after acupuncturing Neiguan (PC 6) by a high sensitive CO2 instrument (Frewil-Q.F., made in China). Mechanical pressure of 1 kg (2 cm～ 1 cm area) was given on Ximen (PC 4) or Daling (PC 7) during the acupuncture and TCE were measured before acupuncture, during acupuncture and mechanical pressing, withdrawing acupuncture but keeping pressing and after withdrawing the pressure. Results: TCE increased significantly on point a, b and Quze (PC 3) during and after acupuncture without mechanical press. When mechanical press was given at Ximen (PC 4), no significantly increase was found on above three points during and after acupuncture. When pressing Daling (PC 7), there was still significant (P<0.05) increase at Quz～ (PC 3) and point during acupuncture and significant increase (P<0.05) at point b after acupuncuture while pressing was kept. Conclusion: Acupuncture can enhance the energy metabolism expressed by TCE along meridians and this effect can be eliminated by mechanical pressing along meridians on the way between acupuncture point and measured point.

Influence of Different Acupuncture Method on Points Transcutaneous CO2 Emission / 针灸推拿医学（英文版）

Objective: To observe the influence of transcutaneous CO2emission (TCE) at points on the meridian or beside the meridian after different stimulations like acupuncture De-qi, sham acupuncture, study the correlation between acupuncture De-qi and local energy metabolism through TCE. Methods: Twenty healthy volunteers (male 8, female 12, average age 29 years old) were selected. Before, during and after five different acupuncture Neignan (PC 6), the TCE was measured at Quze (PC 3), point A [at the lower one fourth of line between Tianquan (PC 2) and Quze (PC 3)], point B [at the middle of line between Ximen (PC 4) and Quze (PC 3)], point C (2 cm ulnar side point A) and point D (2 cm ulnar side point B) with a high sensitive C02 instrument (Frewil-Q.F., made in China). The measure results were compared. Results: The comparison on value of TCE before and after the acupuncture, in Sham acupuncture and shallow acupuncture groups, it increased in all five points; in De-qi acupuncture group, there are significant increase on point A, point B, and Quze (PC 3), it has statistic significance, but there are no significant increase on point C and D. After Ximen (PC 4) or Daling (PC 7) were pressed to block Neiguan (PC 6), the value of TCE on Quze (PC 3), point A and point B were decreased during acupuncture Neiguan (PC 6), and increased after the block was removed, but there was no significant difference. Conclusion: Sham acupuncture has also some effects on body, it can increase energy metabolism of local skin, it may be the reason that shallow acupuncture without qi amving can obtain clinical efficacy. The operation method of traditional qi arriving acupuncture can increase energy metabolism of relevant meridian skin, mechanical press can reduce this effect.

Diuretic effect of injecting furosemide into low hydraulic resistance point Shuifen along the conception meridian in pigs with acute edema / 中西医结合学报

OBJECTIVE: To observe whether injection of medicine into low hydraulic resistance point along meridian brings about higher medicinal effect and to explore the efficacy of the theory that meridians are made up of channels featuring low hydraulic resistance by observing the diuretic effect of injecting furosemide or saline into the low hydraulic resistance point Shuifen (CV 9), vein and Zusanli (St 36) respectively. METHODS: Acute edema was induced in pigs by rapid intravenous injection of 2 000 ml normal saline. The pigs were divided into four groups: Shuifen (CV 9) injection of half dose furosemide group (SFF group), intravenous injection of full dose furosemide group (VF group), Zusanli (St 36) injection of full dose furosemide group (ZSLF group), and Shuifen (CV 9) injection of half dose normal saline group (SFS group). The accumulated urine quantity and the urine quantity generated in every 15-minute period were measured in each group respectively, every 15 minutes after injection, and the measurement lasted for two hours at one experiment. Each group involved eight times of experiments with one pig used for one experiment, which means the whole observation involved 32 times of experiments. RESULTS: The accumulated urine quantities observed in both SFF group and VF group were higher than those in the ZSLF group and the SFS group all through the measurement, showing significant differences during the period from the 15th minute to the 45th minute (P0.05). Analysis of urine quantity generated in every 15-minute period showed that diuretic effect climaxed during the 15th minute to the 30th minute in both SFF group and VF group. By contrast, ZSLF group reached diuresis climax during the 45th minute to 60th minute and no diuresis climax was observed in the SFS group all through the measurement. CONCLUSION: Injection of medicine into low hydraulic resistance point along meridian generates faster and more powerful medicinal potency, and this is likely to be applied to clinical practice. The theory that meridians are channels featuring low hydraulic resistance is important to the elucidation of meridians.