Microbial degradation mechanisms of surface petroleum contaminated seawater in a typical oil trading port.

Petroleum hydrocarbons are significant new persistent organic pollutants for marine oil spill risk areas. Oil trading ports, in turn, have become major bearers of the risk of offshore oil pollution. However, studies on the molecular mechanisms of microbial degradation of petroleum pollutants by natural seawater are limited. Here, an in situ microcosm study was conducted. Combined with metagenomics, differences in metabolic pathways and in the gene abundances of total petroleum hydrocarbons (TPH) are revealed under different conditions. About 88% degradation of TPH was shown after 3 weeks of treatment. The positive responders to TPH were concentrated in the genera Cycloclasticus, Marivita and Sulfitobacter of the orders Rhodobacterales and Thiotrichales. The genera Marivita, Roseobacter, Lentibacter and Glaciecola were key degradation species when mixing dispersants with oil, and all of the above are from the Proteobacteria phylum. The analysis showed that the biodegradability of aromatic compounds, polycyclic aromatic hydrocarbon and dioxin were enhanced after the oil spill, and genes with higher abundances of bphAa, bsdC, nahB, doxE and mhpD were found, but the photosynthesis-related mechanism was inhibited. The dispersant treatment effectively stimulated the microbial degradation of TPH and then accelerated the succession of microbial communities. Meanwhile, functions such as bacterial chemotaxis and carbon metabolism (cheA, fadeJ and fadE) were better developed, but the degradation of persistent organic pollutants such as polycyclic aromatic hydrocarbons was weakened. Our study provides insights into the metabolic pathways and specific functional genes for oil degradation by marine microorganisms and will help improve the application and practice of bioremediation.

Value of Thymosin α1 Combined With Blood Purification to Increase Successful Rescues of Shock Patients.

Context: Septic shock (SS) can pose a high risk of death if rescue efforts in an emergency room aren't started in a timely manner. Thus, rapid and efficient treatment is of great significance to the SS patients' survival. T-α1 can enhance the cellular immune function of patients, and blood purification (BP) can improve the hemodynamics of SS patients by clearing inflammatory mediators in the blood. Objective: The study intended to explore the effects of Thymosin α1 (T-α1) plus blood purification (BP) on SS patients under the emergency green channel (GC), a fast and efficient service system that hospitals provide for acutely and critically ill patients. Design: The research team designed a randomized controlled study. Setting: The study took place in the Emergency Department at the Second Affiliated Hospital of Xi'an Jiaotong University in Xi'an, Shaanxi, China. Participants: Participants were 86 SS patients who came to the hospital for treatment between June 2019 and January 2021. Intervention: The research team numbered the patients in sequence according to the admission time of the patients, and then randomly numbered them by the computer, and assigned participants to an intervention or a control group, with 43 participants in the intervention group receiving T-α1 plus BP therapy and 43 participants in the control group receiving BP treatment only. Outcome Measures: The study measured preparation time before treatment, symptom-onset-to-door (SOTD), duration of shock, length of stay in the intensive care unit (ICU), and incidence of adverse reactions. The study also assessed changes between baseline and postintervention in inflammatory cytokines (ICs), immunological function, and myocardial-function markers. Finally, the research team conducted a one-year follow-up to determine participants' prognostic survival. Results: The groups showed no significant differences in the preparation time before treatment, SOTD, rescue success rate, and incidence of adverse events (P > .05), while the intervention group showed a significantly shorter duration of shock and length stay in the ICU and a significantly higher overall response rate (P < .05). The research team observed significant improvements in the T-lymphocyte subsets, ICs, and myocardial function in both groups postintervention, but the changes in the intervention group were significantly greater (P < .05). Follow-up results showed no significant differences in overall survival between the intervention and control groups (P > .05), but the average LC was significantly higher in the intervention group (P < .05). Conclusions: For SS patients, the combination of T-α1 and BP under the emergency GC can effectively improve their immunological and myocardial function, reduce inflammatory reaction, and prolong their LCs, which provides a greater guarantee of the effectiveness of treatment for SS patients in the future.

Acupuncture for symptoms in menopause transition: a randomized controlled trial.

BACKGROUND: Acupuncture has been used for women during menopause transition, but evidence is limited. OBJECTIVE: We sought to evaluate the efficacy of electroacupuncture on relieving symptoms of women during menopause transition. STUDY DESIGN: We conducted a prospective, multicenter, randomized, participant-blinded trial in China mainland. Subjects were randomized to receive 24 treatment sessions of electroacupuncture at traditional acupoints or sham electroacupuncture at nonacupoints over 8 weeks with 24 weeks' follow-up. Primary outcome was the change from baseline in the total score of Menopause Rating Scale at week 8. Secondary outcomes included the changes from baseline in the average 24-hour hot flash score, the Menopause Rating Scale subscale scores, the total score of Menopause-Specific Quality of Life Questionnaire and its subscales, and serum female hormones. All analyses were performed with a 2-sided P value of < .05 considered significant based on the intention-to-treat principle. RESULTS: A total of 360 women (180 in each group) with menopause-related symptoms during menopause transition were enrolled from June 9, 2013, through Dec 28, 2015. At week 8, the reduction from baseline in the Menopause Rating Scale total score was 6.3 (95% confidence interval, 5.0-7.7) in the electroacupuncture group and 4.5 (95% confidence interval, 3.2-5.8) in the sham electroacupuncture group with a between-group difference of 1.8 (95% confidence interval, 0.9-2.8; P = .0002), less than the minimal clinically important difference of 5 points' reduction. For secondary outcomes, the between-group differences for the decrease in the mean 24-hour hot flash score were significant at weeks 8, 20, and 32, but all were less than the minimal clinically important difference in previous reports. Interestingly, the between-group differences for the Menopause-Specific Quality of Life Questionnaire total score reduction were 5.7 at week 8, 7.1 at week 20, and 8.4 at week 32, greater than the minimal clinically important difference of 4 points. Changes from baseline in follicle-stimulating hormone, luteinizing hormone, and estradiol levels at weeks 8 and 20 (P > .05 for all), with the exception of follicle-stimulating hormone/luteinizing hormone ratios (P = .0024 at week 8 and .0499 at week 20), did not differ between groups. CONCLUSION: Among women during menopause transition, 8 weeks' electroacupuncture treatment did not seem to relieve menopausal symptoms, even though it appeared to improve their quality of life. Generalizability of the trial results may be limited by mild baseline menopausal symptoms in the included participants.

[Effects of electroacupuncture at "Zhongliao" (BL 33) and "Tianshu" (ST 25) on ovarian function in rats with premature ovarian insufficiency].

OBJECTIVE: To explore the effects of electroacupuncture (EA) at "Zhongliao" (BL 33) and "Tianshu" (ST 25) on ovarian function in rats with premature ovarian insufficiency (POI). METHODS: A total of 48 SD female rats with regular estrus were divided into a blank group (n=8), a model group (n=10), an EA group (n=10), a binding group (n=10) and a tamoxifen (TAM) group (n=10). The rats in the model group, EA group, binding group and TAM group were all treated with intraperitoneal injection of 4-vinylcyclohexene diepoxide (VCD, 160 mg/kg) for 15 consecutive days to establish the model of POI; the rats in the blank group were treated with normal diet. After the model was established successfully, the rats in the EA group were treated with EA at "Zhongliao" (BL 33) and "Tianshu" (ST 25) with continuous wave (1 to 3 Hz, 0.1 to 1 mA) for 20 minutes, once a day (five times a week) for the first two weeks and once every other day (three times a week) for the following two weeks. The rats in the TAM group were treated with subcutaneous injection of tamoxifen (1mg/kg), once a day (five times a week) for the first two weeks and once every other day (three times a week) for the following two weeks. The rats in the binding group were bound by a small sack as the EA group. The rats in the blank group and the model group were treated with normal diet. After four weeks, the sexual gland weight and index were tested in each group; the ELISA method was applied to test the level of anti-mllerian hormone (AMH) and inhibin B; the morphology of ovary was observed; the number of primordial follicles, primary follicle, antral follicle and atretic follicle was counted; the expression of insulin-like growth factor-1 (IGF-1) and insulin-like growth factor-1 receptor (IGF-1R) were measured. RESULTS: (1) Compared with the blank group, the ovary weight, ovary index, uterus weight and uterus index were significantly decreased after treatment in the model group, EA group, binding group and TAM group (all P<0.01); but the differences between the model group and the EA group, binding group, TAM group were not significant (all P>0.05). (2) Compared with the blank group, the levels of serum AMH, inhibin B and E2 were significantly reduced; the levels of FSH and LH were significantly increased in the model group; EA group, binding group and TAM group (all P<0.01). Compared with the model group, the levels of serum AMH, inhibin B and E2 were significantly increased, the level of FSH and LH were significantly reduced in the EA group and TAM group (all P<0.01). (3) Compared with the blank group, in the model group, EA group, binding group and TAM group the ovary was dark red and pale, surrounded by particle or not; the morphology was small and atrophic; the primordial follicles was reduced even vanished; the structure of primary follicle was damaged and loosely arranged; the mature follicle was few; the atretic follicle and interstitial gland were increased. (4) Compared with the blank group, the expressions of IGF-1 mRNA and IGF-1R mRNA were increased in the model group (all P<0.01); compared with the blank group, the expression of IGF-1 mRNA was increased in the binding group (P<0.05), but that of IGF-1R mRNA was not significantly different (P>0.05); compared with the model group, the expression of IGF-1 mRNA was not significantly different in the EA group, binding group and TAM group (all P>0.05), but that of IGF-1R mRNA was reduced (P<0.05, P<0.01). CONCLUSION: EA at "Zhongliao" (BL 33) and "Tianshu" (ST 25) has improvement effect on ovarian function in rats with VCD-induced POI, which is likely to be related to regulating the IGF-1R mRNA expression to improve the IGF-1/ IGF-1R axis.

[Protective effects of electroacupuncture pretreatment on ovarian in rats with premature ovarian insufficiency].

OBJECTIVE: To explore the protective effect and possible mechanism of electroacupuncture (EA) pretreatment at "Zhongliao" (BL 33) and "Tianshu" (ST 25) on ovarian in rats with premature ovarian insufficiency (POI). METHODS: A total of 32 SD female rats with regular estrus were randomly divided into a blank group, a model group, an EA group and an immobilization group, 8 rats in each one. The rats in the model group, EA group and immobilization group were treated with intraperitoneal injection of 4-vinylcyclohexene diepoxide (160 mg/kg) for 15 days to establish the model. From the beginning of model establishment, the rats in the EA group were treated with EA pretreatment at "Zhongliao" (BL 33) and "Tianshu" (ST 25), once a day for the first two weeks (5 treatments per week) and once every two days for the last two weeks (3 treatments per week). EA was given 20 min per treatment for four weeks. The rats in the immobilization group were treated with immobilization with identical duration, frequency and total amount as the EA group. No treatment was given in the blank group and model group. When model was successfully established in the model group, rats were sacrificed to test the weight of sexual gland and the sexual gland index; the ELISA method was applied to test the level of anti-müllerian hormone (AMH) and inhibin B; the radioimmunoassay method was applied to test the level of follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), progesterone (P) and testosterone (T). In addition, the morphology and the number of ovarian follicles were observed; and western blot method was applied to test the expression of B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax) in ovarian tissue. RESULTS: Compared with the blank group, the bilateral ovary weight, ovary index, uterus weight and uterus index decreased (P<0.05, P<0.01); serum AMH, inhibin B, E2 and P reduced, while serum FSH, LH and T increased in the model group (all P<0.01); primordial follicles and primary follicles decreased, while atretic follicles increased in the model group (all P<0.01); the expression of Bax protein increased, while the expression of Bcl-2 protein decreased in the model group (both P<0.01). Compared with the model group, the bilateral ovary weight, ovary index and uterus weight increased (P<0.05, P<0.01); serum AMH, inhibin B, E2 and P increased, while serum FSH and LH reduced (P<0.05, P<0.01); primordial follicles and primary follicles increased, while atretic follicles reduced (P<0.05, P<0.01); the expression of Bax protein reduced, while the expression of Bcl-2 protein increased in the EA group (P<0.05, P<0.01). The differences of each outcome indexes were not significant between the model group and immobilization group (all P>0.05). CONCLUSION: EA pretreatment at "Zhongliao" (BL 33) and Tianshu (ST 25) for 4 weeks has certain protective effect on ovarian in POI rats, which is likely to be related with up-regulating ovarian granular cells, inhabiting the expression of Bcl-2 protein and down-regulating the expression of Bax protein.

A comparative study of electroacupuncture at Zhongliao (BL33) and other acupoints for overactive bladder symptoms.

Electroacupuncture (EA) at Zhongliao (BL33) can improve the symptoms of overactive bladder (OAB), such as urinary frequency, urgency, and incontinence. However, its performance compared with other acupoints remains unclear. This study investigated the effects of EA at BL33 with deep needling on rats with OAB by detecting urodynamics in eight groups: no intervention group, D-BL33 group (deep needling at BL33), S-BL33 group (shallow needling at BL33), non-acupoint group (needling at the non-acupoint next to BL33), Weizhong (BL40) group, Sanyinjiao (SP6) group, Tongtian (BL7) group, and Hegu (LI4) group. Results revealed that EA at BL33 with deep needling, BL40, and SP6 prolonged the intercontraction interval (ICI) of rats with OAB (P = 0.001, P = 0.005, P = 0.046, respectively, post-treatment vs. post-modeling). Furthermore, the change in ICI from post-modeling in the D-BL33 group was significantly greater than those of the no intervention and other EA groups (all P < 0.01). Significantly shortened vesical micturition time (VMT) and elevated maximum detrusor pressure (MDP) were also observed in the D-BL33 group (P = 0.017 and P = 0.024, respectively, post-treatment vs. post-modeling). However, no statistically significant differences in the changes of VMT and MDP from post-modeling were observed between D-BL33 and the other EA groups. In conclusion, EA at BL33 with deep needling may inhibit acetic-acid-induced OAB more effectively.

Efficacy of acupuncture on menstrual frequency in women with polycystic ovary syndrome: Protocol for a randomized, controlled trial.

BACKGROUND: Acupuncture may improve the menstrual frequency of women with polycystic ovary syndrome (PCOS). However, more sufficient data are needed to improve the efficacy of acupuncture. METHODS: A total of 172 participants diagnosed with PCOS would be randomly assigned to either the acupuncture group or sham-acupuncture group, at a ratio of 1:1. Participants in both groups will receive treatment for 12 weeks, 3 times a week. The primary outcome will be the proportion of participants with at least a 50% increase from baseline in the monthly menstrual frequency from baseline after 12 weeks intervention, while secondary outcomes will be the difference in anthropometrics, serum hormone level, ovarian morphology, anxiety and depression, and quality of life from baseline to after 12 weeks intervention and to 12 weeks postintervention follow-up between groups. DISCUSSION: The aim of this study is to evaluate the efficacy and safety of acupuncture for improving menstrual frequency and other symptoms of patients with PCOS. The limitation of this trial is that it would be difficult to blind the acupuncturists. In addition, these findings may not be suitable for women with PCOS who are seeking pregnancy.

Efficacy of acupuncture for degenerative lumbar spinal stenosis: protocol for a randomised sham acupuncture-controlled trial.

INTRODUCTION: Degenerative lumbar spinal stenosis (DLSS) is a major public health problem and the primary reason why older adults seek lumbar spine surgery. Acupuncture may be effective for DLSS, but the evidence supporting this possibility is still limited. METHODS AND ANALYSIS: A total of 80 participants with DLSS will be randomly allocated to either an acupuncture group or a sham acupuncture (SA) group at a ratio of 1:1. 24 treatments will be provided over 8 weeks. The primary outcome is the score change of the Modified Roland-Morris Disability Questionnaire (RMDQ) responses from baseline to week 8. The secondary outcomes include the assessment of lower back pain and leg pain using the Numeric Rating Scale (NRS), the change in the number of steps per month, and the assessment of the specific quality of life using the Swiss Spinal Stenosis Questionnaire (SSSQ). We will follow-up with the participants until week 32. All of the participants who received allocation will be included in the statistical analysis. ETHICS/DISSEMINATION: This protocol has been approved by the Research Ethical Committee of Guang'anmen Hospital (Permission number: 2015EC114) and Fengtai Hospital of Integrated Traditional and Western Medicine (Permission number: 16KE0409). The full data set will be made available when this trial is completed and published. Applications for the release of data should be made to ZL (principal investigator). TRIAL REGISTRATION NUMBER: NCT02644746.

Atorvastatin ameliorates early brain injury after subarachnoid hemorrhage via inhibition of AQP4 expression in rabbits.

The therapeutic effects of atorvastatin on early brain injury (EBI), cerebral edema and its association with aquaporin 4 (AQP4) were studied in rabbits after subarachnoid hemorrhage (SAH) using western blot analysis and the dry-wet method. Seventy-two healthy male New Zealand rabbits weighing between 2.5 and 3.2 kg were randomly divided into three groups: the SAH group (n=24), sham-operated group (n=24) and the SAH + atorvastatin group (n=24). A double SAH model was employed. The sham-operated group were injected with the same dose of saline solution, the SAH + atorvastatin group received atorvastatin 20 mg/kg/day after SAH. All rabbit brain samples were taken at 72 h after the SAH model was established successfully. Brain edema was detected using the dry-wet method after experimental SAH was induced; AQP4 and caspase-3 expression was measured by western blot analysis, and neuronal apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) staining at 72 h after SAH. The results indicated that brain edema and injury appeared soon after SAH, while brain edema and EBI were ameliorated and increased behavior scores were noted after prophylactic use of atorvastatin. Compared with the SAH group, the level of AQP4 and the cerebral content of water was significantly decreased (P<0.01) by atorvastatin, and TUNEL staining and studying the expression of caspase-3 showed that the apoptosis of neurons was reduced markedly both in the hippocampus and brain cortex by atorvastatin. The results suggest that atorvastatin ameliorated brain edema and EBI after SAH, which was related to its inhibition of AQP4 expression. Our findings provide evidence that atorvastatin is an effective and well-tolerated approach for treating SAH in various clinical settings.

A case study of an enhanced eutrophication model with stoichiometric zooplankton growth sub-model calibrated by Bayesian method.

Urban lakes in China have suffered from severe eutrophication over the past several years, particularly those with relatively small areas and closed watersheds. Many efforts have been made to improve the understanding of eutrophication physiology with advanced mathematical models. However, several eutrophication models ignore zooplankton behavior and treat zooplankton as particles, which lead to the systematic errors. In this study, an eutrophication model was enhanced with a stoichiometric zooplankton growth sub-model that simulated the zooplankton predation process and the interplay among nitrogen, phosphorus, and oxygen cycles. A case study in which the Bayesian method was used to calibrate the enhanced eutrophication model parameters and to calculate the model simulation results was carried out in an urban lake in Tianjin, China. Finally, a water quality assessment was also conducted for eutrophication management. Our result suggests that (1) integration of the Bayesian method and the enhanced eutrophication model with a zooplankton feeding behavior sub-model can effectively depict the change in water quality and (2) the nutrients resulting from rainwater runoff laid the foundation for phytoplankton bloom.

Impact of partial reimbursement on hepatitis B antiviral utilization and adherence.

AIM: To determine the impact of partial reimbursement for antivirals on antiviral utilization and adherence for chronic hepatitis B patients. METHODS: This was a retrospective cohort study. Two separate cohorts were enrolled, including 14163 and 16288 chronic hepatitis B outpatients, respectively. These patients were referred to Beijing You'an Hospital before and after the new partial reimbursement for antivirals, which was implemented on July 1, 2011. Demographic characteristics (including medical insurance status), routine biochemical, virological and serology laboratory test results, and antiviral agents' prescription information were collected from an electronic database. Patients were also defined as new and existing patients according to treatment history. Antiviral utilization, medication possession ratio and persistence rate were calculated and compared among the patients with different characteristics. A questionnaire survey was conducted among 212 randomly sampled outpatients from the same hospital to confirm the validity of the electronic database. Propensity score matching was used to adjust the distribution of patient's characteristics which may influence the antiviral utilization. χ(2) test or ANOVA was adopted and multivariate logistic regression was used to determine the factors associated with antiviral utilization and good adherence. RESULTS: A total of 13364 outpatients from each cohort were enrolled after the propensity score matching. The antiviral utilization rate for the insured patients increased from 57.4% to 75.9% (P < 0.0001) after the reimbursement, and the rate among those who paid out-of-pocket increased from 54.9% to 56.7% (P = 0.028). Approximately 71% of the patients had a medication possession ratio of more than 80% in each cohort before reimbursement. This increased to 79.2% and 73.1% for insured patients and those who paid out-of-pocket, respectively (P < 0.0001). Insured patients and those who paid out-of-pocket had the similar persistence rates before reimbursement. But after reimbursement, insured patients had higher persistence rates than those who paid out-of-pocket at 6 (86.5% vs 81.5%, P < 0.0001), 9 (79.7% vs 69.9%, P < 0.0001), 12 (73.4% vs 61.9%, P < 0.0001), and 15 mo (66.6% vs 53.1%, P < 0.0001). The reimbursement could significantly improve adherence for the insured patients than those who paid out-of-pocket even after adjusting other covariates, with an interaction odds ratio of 1.422 (95%CI: 1.220-1.657, P < 0.0001). The questionnaire survey supported the validity of the electronic database. CONCLUSION: The reimbursement policy shows a positive impact on antiviral utilization as well as adherence for insured chronic hepatitis B patients.

Identification and Characterization of a PRDM14 Homolog in Japanese Flounder (Paralichthys olivaceus).

PRDM14 is a PR (PRDI-BF1-RIZ1 homologous) domain protein with six zinc fingers and essential roles in genome-wide epigenetic reprogramming. This protein is required for the establishment of germ cells and the maintenance of the embryonic stem cell ground state. In this study, we cloned the full-length cDNA and genomic DNA of the Paralichthys olivaceus prdm14 (Po-prdm14) gene and isolated the 5' regulatory region of Po-prdm14 by whole-genome sequencing. Peptide sequence alignment, gene structure analysis, and phylogenetic analysis revealed that Po-PRDM14 was homologous to mammalian PRDM14. Results of real-time quantitative polymerase chain reaction amplification (RT-qPCR) and in situ hybridization (ISH) in embryos demonstrated that Po-prdm14 was highly expressed between the morula and late gastrula stages, with its expression peaking in the early gastrula stage. Relatively low expression of Po-prdm14 was observed in the other developmental stages. ISH of gonadal tissues revealed that the transcripts were located in the nucleus of the oocytes in the ovaries but only in the spermatogonia and not the spermatocytes in the testes. We also presume that the Po-prdm14 transcription factor binding sites and their conserved binding region among vertebrates. The combined results suggest that Po-PRDM14 has a conserved function in teleosts and mammals.

Integration of Bayesian analysis for eutrophication prediction and assessment in a landscape lake.

Eutrophication models have been widely used to assess water quality in landscape lakes. Because flow rate in landscape lakes is relatively low and similar to that of natural lakes, eutrophication is more dominant in landscape lakes. To assess the risk of eutrophication in landscape lakes, a set of dynamic equations was developed to simulate lake water quality for total nitrogen (TN), total phosphorous (TP), dissolve oxygen (DO) and chlorophyll a (Chl a). Firstly, the Bayesian calibration results were described. Moreover, the ability of the model to reproduce adequately the observed mean patterns and major cause-effect relationships for water quality conditions in landscape lakes were presented. Two loading scenarios were used. A Monte Carlo algorithm was applied to calculate the predicated water quality distributions, which were used in the established hierarchical assessment system for lake water quality risk. The important factors affecting the lake water quality risk were defined using linear regression analysis. The results indicated that the variations in the landscape lake receiving recharge water quality caused considerable landscape lake water quality risk in the surrounding area. Moreover, the Chl a concentration in lake water was significantly affected by TP and TN concentrations; the lake TP concentration was the limiting factor for growth of plankton in lake water. The lake water TN concentration provided the basic nutritional requirements. Lastly, lower TN and TP concentrations in the receiving recharge water caused increased lake water quality risk.

Effects of different calcium, magnesium, and zinc concentrations supplemented on hepatopancreatic cell proliferation of kuruma prawn, Penaeus japonicus.

The effects of different calcium (Ca(2+)), magnesium (Mg(2+)), and zinc (Zn(2+)) concentrations supplemented on hepatopancreatic cell proliferation of kuruma prawn, Penaeus japonicus was studied. The culture system consists of medium 199 (M 199) supplemented with 0.060 mol/L NaCl, 1.011 g/L glucose, 1,000 UI/ml penicillin, 1,000 µg/ml treptomycin, heat inactivated fetal calf serum (FCS) 20% for primary cell culture and 10% for subculture. The RNA/DNA ratio of the cell cultures was measured. The results show that the cell division of hepatopancreatic cells of P. japonicus was enhanced by the optimal concentration of inorganic salt (Ca(2+), 1.0 g/L; Mg(2+), 5.0 g/L; Zn(2+), 80 µg/L). The hepatopancreatic cell culture system and improved culture conditions described here will be very useful for in vitro experiments to study viruses responsible for infections in shrimp leading to tremendous economic losses.

[Effects of Yanggan Lidan Granule on insulin resistance in guinea pigs with induced cholesterol gallstones].

OBJECTIVE: To observe the effects of Yanggan Lidan Granule (YGLDG), a compound traditional Chinese herbal medicine, on insulin resistance in guinea pigs with induced cholesterol gallstones. METHODS: Eighty guinea pigs were randomly divided into normal control group, untreated group, YGLDG group and ursodeoxycholic acid (UDCA) group, with 20 guinea pigs in each group. Except the normal control group, gallstones were induced by high-cholesterol diet in the guinea pigs. The guinea pigs in the normal control group and the untreated group were administered with normal saline. UDCA and YGLDG were given to the guinea pigs in the corresponding groups for seven weeks. Eight guinea pigs of each group were used to measure the glucose infusion rate (GIR) by using hyperinsulinemic-euglycemic clamp technique. At the end the guinea pigs were killed and their gallstone formation was observed. RESULTS: The gallstones in guinea pigs were identified as cholesterol stones by qualitative analysis through infrared spectrum. The incidence rate of cholelithiasis of the untreated group was 82.35% . The GIR of guinea pigs in the untreated group was obviously lowered down as compared with the normal control group. Compared with the untreated group, the GIRs of the YGLDG group and the UDCA group were obviously increased, especially in the YGLDG group. CONCLUSION: YGLDG may improve insulin resistance in guinea pigs with cholesterol gallstones by elevating GIR obviously.

[Clinical observation on effects of routine acupuncture combined with Feiteng Bafa for chloasma].

OBJECTIVE: To explore the relatively efficacious acupuncture treatment for chloasma, to promote its clinical application in medical settings. METHODS: Forty cases were randomly divided into a Feiteng Bafa group (FB) and a selecting points based on differentiation of syndromes group (DS), 20 cases in each group and acupoints. Hegu (LI 4), acupoints in close proximity to chloasma and according to differentiation of syndrome were selected in DS group. The acupoints selected according to Feiteng Bafa principles were adhered in FB group. After two treatment courses, the effectiveness rate, the pro-and post-size and pro-and post-color of the chloasma area were compared between the two groups. RESULTS: The size of the chloasma area decreased, and the color of the chloasma area became lighter in both groups (P < 0.01, P < 0.05), and the FB group was better than the DS group (both P < 0.05). The effectiveness rate in the FB group was 95.0%, better than that of 70.0% in the DS group. CONCLUSION: The acupuncture treatment adhered to Feiteng Bafa is better than the simple routine acupuncture for chloasma.